The	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
type	I-Gene_or_gene_product
II	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
in	O
chronic	B-Cancer
myeloid	I-Cancer
leukemia	I-Cancer
:	O
analysis	O
of	O
microsatellite	O
regions	O
and	O
gene	O
expression	O
.	O

Genomic	O
instability	O
is	O
one	O
mechanism	O
proposed	O
to	O
play	O
a	O
role	O
in	O
the	O
disease	O
progression	O
of	O
chronic	B-Cancer
myeloid	I-Cancer
leukemia	I-Cancer
(	O
CML	B-Cancer
)	O
.	O

Microsatellite	O
regions	O
in	O
the	O
type	B-Gene_or_gene_product
II	I-Gene_or_gene_product
transforming	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
(	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
RII	I-Gene_or_gene_product
)	O
gene	O
appear	O
to	O
be	O
targets	O
for	O
mutation	O
in	O
some	O
cancers	B-Cancer
displaying	O
microsatellite	O
instability	O
(	O
replication	O
error	O
phenotype	O
,	O
RER	O
+	O
)	O
.	O

Furthermore	O
,	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
RII	I-Gene_or_gene_product
mutations	O
in	O
RER	B-Cancer
+	I-Cancer
tumors	I-Cancer
have	O
been	O
associated	O
with	O
decreased	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
RII	I-Gene_or_gene_product
mRNA	O
levels	O
.	O

As	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
is	O
a	O
potent	O
negative	O
growth	O
regulator	O
of	O
hematopoietic	B-Cell
cells	I-Cell
,	O
investigations	O
were	O
undertaken	O
to	O
determine	O
whether	O
inactivation	O
of	O
the	O
receptor	O
by	O
microsatellite	O
alteration	O
might	O
be	O
involved	O
in	O
the	O
progression	O
of	O
CML	B-Cancer
.	O

Analysis	O
of	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
RII	I-Gene_or_gene_product
mRNA	O
expression	O
by	O
RNase	B-Gene_or_gene_product
protection	O
,	O
with	O
comparison	O
of	O
cells	B-Cell
from	O
the	O
chronic	O
,	O
accelerated	O
and	O
blast	O
phases	O
of	O
CML	B-Cancer
,	O
showed	O
no	O
change	O
in	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
RII	I-Gene_or_gene_product
transcript	O
levels	O
during	O
disease	O
progression	O
.	O

However	O
,	O
during	O
each	O
phase	O
of	O
the	O
disease	O
,	O
low	O
levels	O
of	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
RII	I-Gene_or_gene_product
were	O
detected	O
when	O
compared	O
with	O
the	O
hematopoietic	B-Cell
cells	I-Cell
of	O
normal	O
donors	B-Organism
.	O

Furthermore	O
,	O
this	O
decreased	O
expression	O
was	O
also	O
observed	O
in	O
the	O
other	O
myeloproliferative	O
disorders	O
,	O
polycythemia	O
rubra	O
vera	O
(	O
PRV	O
)	O
and	O
essential	O
thrombocythemia	O
(	O
ET	O
)	O
.	O

The	O
leukemia	B-Cell
cell	I-Cell
lines	I-Cell
K562	I-Cell
and	O
HL	B-Cell
-	I-Cell
60	I-Cell
had	O
no	O
detectable	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
RII	I-Gene_or_gene_product
mRNA	O
.	O

Two	O
microsatellite	O
regions	O
found	O
altered	O
in	O
RER	B-Cancer
+	I-Cancer
colon	I-Cancer
cancers	I-Cancer
were	O
analyzed	O
to	O
establish	O
if	O
these	O
sequences	O
were	O
aberrant	O
in	O
CML	B-Cancer
.	O

No	O
alteration	O
was	O
detected	O
in	O
either	O
of	O
these	O
regions	O
in	O
any	O
phase	O
of	O
the	O
disease	O
.	O

These	O
results	O
suggest	O
that	O
alterations	O
of	O
the	O
microsatellite	O
regions	O
in	O
the	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
RII	I-Gene_or_gene_product
gene	O
are	O
not	O
involved	O
in	O
the	O
progression	O
of	O
CML	B-Cancer
.	O

Decreased	O
expression	O
of	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
RII	I-Gene_or_gene_product
in	O
CML	B-Cell
cells	I-Cell
and	O
leukemia	B-Cell
cell	I-Cell
lines	I-Cell
raises	O
the	O
possibility	O
that	O
altered	O
expression	O
of	O
the	O
receptor	O
may	O
play	O
a	O
role	O
in	O
the	O
initiation	O
and	O
/	O
or	O
maintenance	O
of	O
the	O
disease	O
state	O
.	O

N	O
-	O
glycosylation	O
of	O
glucose	B-Gene_or_gene_product
transporter	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
Glut	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
is	O
associated	O
with	O
increased	O
transporter	O
affinity	O
for	O
glucose	B-Simple_chemical
in	O
human	B-Organism
leukemic	B-Cell
cells	I-Cell
.	O

To	O
elucidate	O
the	O
role	O
of	O
N	O
-	O
glycosylation	O
in	O
the	O
functional	O
activity	O
of	O
the	O
universal	O
glucose	B-Simple_chemical
transporter	O
,	O
Glut	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
we	O
investigated	O
effects	O
of	O
the	O
N	O
-	O
glycosylation	O
inhibitor	O
,	O
tunicamycin	B-Simple_chemical
,	O
on	O
glucose	B-Simple_chemical
transport	O
by	O
human	B-Organism
leukemic	B-Cell
cell	I-Cell
lines	I-Cell
K562	I-Cell
,	O
U937	B-Cell
and	O
HL60	B-Cell
.	O

Treatment	O
with	O
tunicamycin	B-Simple_chemical
produced	O
a	O
40	O
-	O
50	O
%	O
inhibition	O
of	O
2	B-Simple_chemical
-	I-Simple_chemical
deoxyglucose	I-Simple_chemical
uptake	O
and	O
this	O
was	O
associated	O
with	O
a	O
2	O
-	O
2	O
.	O
5	O
-	O
fold	O
decrease	O
in	O
transporter	O
affinity	O
for	O
glucose	B-Simple_chemical
(	O
Km	O
)	O
without	O
a	O
change	O
in	O
Vmax	O
.	O

Leukemic	B-Cell
K562	I-Cell
,	O
U937	B-Cell
and	O
HL60	B-Cell
cells	I-Cell
expressed	O
Glut	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
transporter	O
protein	O
.	O

With	O
K562	B-Cell
cells	I-Cell
Glut	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
appeared	O
as	O
a	O
broad	O
band	O
of	O
50	O
-	O
60	O
kDa	O
,	O
whereas	O
with	O
U937	B-Cell
and	O
HL60	B-Cell
cells	I-Cell
a	O
diffuse	O
band	O
was	O
observed	O
at	O
approximately	O
55	O
kDa	O
.	O

Treatment	O
of	O
K562	B-Cell
cells	I-Cell
with	O
tunicamycin	B-Simple_chemical
for	O
18	O
h	O
,	O
resulted	O
in	O
extensive	O
loss	O
of	O
the	O
50	O
-	O
60	O
kDa	O
glycoprotein	B-Gene_or_gene_product
,	O
appearance	O
of	O
a	O
30	O
-	O
40	O
kDa	O
band	O
and	O
increased	O
staining	O
of	O
a	O
45	O
kDa	O
band	O
.	O

With	O
U937	B-Cell
cells	I-Cell
,	O
tunicamycin	B-Simple_chemical
treatment	O
resulted	O
in	O
the	O
appearance	O
of	O
a	O
30	O
-	O
40	O
kDa	O
band	O
and	O
increased	O
staining	O
of	O
a	O
45	O
kDa	O
band	O
.	O

With	O
HL60	B-Cell
cells	I-Cell
loss	O
of	O
the	O
55	O
kDa	O
Glut	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
band	O
was	O
observed	O
and	O
a	O
band	O
of	O
45	O
kDa	O
appeared	O
.	O

Tunicamycin	B-Simple_chemical
-	O
treatment	O
resulted	O
in	O
75	O
-	O
90	O
%	O
inhibition	O
in	O
[	B-Simple_chemical
3H	I-Simple_chemical
]	I-Simple_chemical
mannose	I-Simple_chemical
incorporation	O
but	O
only	O
20	O
-	O
25	O
%	O
inhibition	O
in	O
[	B-Simple_chemical
3H	I-Simple_chemical
]	I-Simple_chemical
thymidine	I-Simple_chemical
and	O
[	B-Simple_chemical
3H	I-Simple_chemical
]	I-Simple_chemical
leucine	I-Simple_chemical
incorporation	O
.	O

In	O
contrast	O
,	O
tunicamycin	B-Simple_chemical
had	O
little	O
effect	O
on	O
the	O
viability	O
and	O
MTT	B-Simple_chemical
responses	O
of	O
the	O
cells	B-Cell
used	O
.	O

These	O
results	O
suggest	O
that	O
in	O
leukemic	B-Cell
cells	I-Cell
N	O
-	O
glycosylation	O
of	O
Glut	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
plays	O
an	O
important	O
role	O
in	O
maintaining	O
its	O
structure	O
and	O
functional	O
integration	O
.	O

Immunohistochemical	O
analysis	O
of	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
gene	O
product	O
in	O
node	B-Multi-tissue_structure
-	O
positive	O
lung	B-Cancer
cancer	I-Cancer
and	O
lymph	B-Multi-tissue_structure
nodes	I-Multi-tissue_structure
.	O

The	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
gene	O
product	O
has	O
been	O
considered	O
as	O
an	O
anti	O
-	O
metastatic	O
protein	O
and	O
the	O
level	O
of	O
its	O
expression	O
has	O
been	O
reported	O
to	O
correlate	O
inversely	O
with	O
metastatic	O
potential	O
in	O
some	O
cancers	B-Cancer
.	O

However	O
,	O
the	O
expression	O
of	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
gene	O
product	O
in	O
the	O
metastatic	O
sites	B-Multi-tissue_structure
have	O
not	O
been	O
studied	O
in	O
detail	O
.	O

We	O
examined	O
the	O
expression	O
of	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
gene	O
product	O
in	O
surgically	O
resected	O
46	O
pairs	O
of	O
primary	O
lung	B-Cancer
cancers	I-Cancer
and	O
metastatic	O
lymph	B-Multi-tissue_structure
nodes	I-Multi-tissue_structure
by	O
immunohistochemistry	O
.	O

The	O
positive	O
staining	O
of	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
gene	O
product	O
in	O
primary	B-Cancer
cancers	I-Cancer
and	O
metastatic	O
lymph	B-Multi-tissue_structure
nodes	I-Multi-tissue_structure
were	O
observed	O
in	O
56	O
.	O
5	O
and	O
67	O
.	O
4	O
%	O
,	O
respectively	O
.	O

The	O
heterogeneity	O
of	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
gene	O
product	O
expression	O
between	O
primary	B-Cancer
cancers	I-Cancer
and	O
metastatic	O
lymph	B-Multi-tissue_structure
nodes	I-Multi-tissue_structure
was	O
observed	O
in	O
41	O
.	O
3	O
%	O
.	O

No	O
correlations	O
were	O
found	O
between	O
the	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
gene	O
product	O
expression	O
in	O
lung	B-Cancer
cancers	I-Cancer
and	O
the	O
patients	B-Organism
survival	O
.	O

No	O
significant	O
association	O
was	O
also	O
observed	O
between	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
gene	O
product	O
expression	O
in	O
lymph	B-Multi-tissue_structure
nodes	I-Multi-tissue_structure
and	O
the	O
patients	B-Organism
survival	O
.	O

There	O
was	O
,	O
furthermore	O
,	O
no	O
correlation	O
between	O
the	O
heterogeneity	O
of	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
gene	O
product	O
expression	O
and	O
the	O
patients	B-Organism
survival	O
.	O

In	O
conclusion	O
,	O
the	O
level	O
of	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
gene	O
product	O
expression	O
does	O
not	O
significantly	O
reveal	O
prognostic	O
value	O
in	O
node	B-Cancer
-	I-Cancer
positive	I-Cancer
lung	I-Cancer
cancers	I-Cancer
.	O

Expression	O
of	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
gene	O
product	O
in	O
metastatic	O
lymph	B-Multi-tissue_structure
nodes	I-Multi-tissue_structure
was	O
also	O
unrelated	O
to	O
patients	B-Organism
survival	O
.	O

Bradycardia	O
-	O
induced	O
coronary	B-Multi-tissue_structure
angiogenesis	O
is	O
dependent	O
on	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
.	O

A	O
marked	O
coronary	B-Multi-tissue_structure
angiogenesis	O
is	O
known	O
to	O
occur	O
with	O
chronic	O
bradycardia	O
.	O

We	O
tested	O
the	O
hypothesis	O
that	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
,	O
an	O
endothelial	B-Cell
cell	I-Cell
mitogen	O
and	O
a	O
major	O
regulator	O
of	O
angiogenesis	O
,	O
is	O
upregulated	O
in	O
response	O
to	O
low	O
heart	B-Organ
rate	O
and	O
consequential	O
increased	O
stroke	O
volume	O
.	O

Bradycardia	O
was	O
induced	O
in	O
rats	B-Organism
by	O
administering	O
the	O
bradycardic	O
drug	O
alinidine	B-Simple_chemical
(	O
3	O
mg	O
/	O
kg	O
body	B-Organism_subdivision
weight	O
)	O
twice	O
daily	O
.	O

Heart	B-Organ
rate	O
decreased	O
by	O
32	O
%	O
for	O
20	O
to	O
40	O
minutes	O
after	O
injection	O
and	O
was	O
chronically	O
reduced	O
by	O
10	O
%	O
,	O
14	O
%	O
,	O
and	O
18	O
.	O
5	O
%	O
after	O
1	O
,	O
2	O
,	O
and	O
3	O
weeks	O
of	O
treatment	O
,	O
respectively	O
.	O

Arterial	B-Multi-tissue_structure
pressure	O
and	O
cardiac	B-Organ
output	O
were	O
unchanged	O
.	O

Left	B-Tissue
ventricular	I-Tissue
capillary	I-Tissue
length	O
density	O
(	O
mm	O
/	O
mm	O
(	O
3	O
)	O
)	O
increased	O
gradually	O
with	O
alinidine	B-Simple_chemical
administration	O
;	O
a	O
15	O
%	O
increase	O
after	O
2	O
weeks	O
and	O
a	O
40	O
%	O
increase	O
after	O
3	O
weeks	O
of	O
alinidine	B-Simple_chemical
treatment	O
were	O
documented	O
.	O

Left	B-Multi-tissue_structure
ventricular	I-Multi-tissue_structure
weight	O
,	O
body	B-Organism_subdivision
weight	O
,	O
and	O
their	O
ratio	O
were	O
not	O
significantly	O
altered	O
by	O
alinidine	B-Simple_chemical
treatment	O
.	O

After	O
1	O
week	O
of	O
treatment	O
,	O
before	O
an	O
increase	O
in	O
capillary	B-Tissue
length	O
density	O
,	O
VEGF	B-Gene_or_gene_product
mRNA	O
increased	O
greater	O
than	O
2	O
-	O
fold	O
and	O
then	O
declined	O
to	O
control	O
levels	O
after	O
3	O
weeks	O
of	O
treatment	O
.	O

VEGF	B-Gene_or_gene_product
protein	O
was	O
higher	O
in	O
alinidine	B-Simple_chemical
-	O
treated	O
rats	B-Organism
than	O
in	O
controls	O
after	O
2	O
weeks	O
and	O
increased	O
further	O
after	O
3	O
weeks	O
of	O
treatment	O
.	O

Injection	O
of	O
VEGF	B-Gene_or_gene_product
-	O
neutralizing	O
antibodies	O
over	O
a	O
2	O
-	O
week	O
period	O
completely	O
blocked	O
alinidine	B-Simple_chemical
-	O
stimulated	O
angiogenesis	O
.	O

In	O
contrast	O
,	O
bFGF	B-Gene_or_gene_product
mRNA	O
was	O
not	O
altered	O
by	O
alinidine	B-Simple_chemical
treatment	O
.	O

These	O
data	O
suggest	O
that	O
VEGF	B-Gene_or_gene_product
plays	O
a	O
key	O
role	O
in	O
the	O
angiogenic	O
response	O
that	O
occurs	O
with	O
chronic	O
bradycardia	O
.	O

The	O
mechanism	O
underlying	O
this	O
VEGF	B-Gene_or_gene_product
-	O
associated	O
angiogenesis	O
may	O
be	O
an	O
increase	O
in	O
stretch	O
due	O
to	O
enhanced	O
diastolic	O
filling	O
.	O

External	O
beam	O
radiotherapy	O
for	O
subretinal	B-Immaterial_anatomical_entity
neovascularization	O
in	O
age	O
-	O
related	O
macular	B-Tissue
degeneration	O
:	O
is	O
this	O
treatment	O
efficient	O
?	O

PURPOSE	O
:	O
Control	O
of	O
the	O
natural	O
course	O
of	O
subretinal	B-Immaterial_anatomical_entity
neovascularization	O
(	O
SRNV	O
)	O
in	O
age	O
-	O
related	O
macular	B-Tissue
degeneration	O
(	O
AMD	O
)	O
is	O
difficult	O
.	O

Only	O
a	O
subset	O
of	O
patients	B-Organism
is	O
suitable	O
for	O
laser	O
coagulation	O
.	O

This	O
prospective	O
study	O
aimed	O
to	O
determine	O
the	O
efficacy	O
and	O
individual	O
benefit	O
of	O
external	O
beam	O
radiotherapy	O
(	O
EBRT	O
)	O
.	O

METHODS	O
AND	O
MATERIALS	O
:	O
The	O
prospective	O
trial	O
included	O
287	O
patients	B-Organism
with	O
subfoveal	O
neovascularization	O
due	O
to	O
AMD	O
which	O
was	O
verified	O
by	O
fluorescein	B-Simple_chemical
angiography	O
.	O

Patients	B-Organism
have	O
been	O
treated	O
between	O
January	O
1996	O
and	O
October	O
1997	O
.	O

All	O
patients	B-Organism
received	O
a	O
total	O
dose	O
of	O
16	O
Gy	O
in	O
2	O
-	O
Gy	O
daily	O
fractions	O
with	O
5	O
-	O
6	O
MeV	O
photons	O
based	O
on	O
computerized	O
treatment	O
planning	O
in	O
individual	O
head	B-Organism_subdivision
mask	O
fixation	O
.	O

This	O
first	O
analysis	O
is	O
based	O
on	O
73	O
patients	B-Organism
(	O
50	O
women	B-Organism
,	O
23	O
men	B-Organism
,	O
median	O
age	O
74	O
.	O
3	O
years	O
)	O
,	O
with	O
a	O
median	O
follow	O
-	O
up	O
of	O
13	O
.	O
3	O
months	O
and	O
a	O
minimum	O
follow	O
-	O
up	O
of	O
11	O
months	O
.	O

RESULTS	O
:	O
All	O
patients	B-Organism
completed	O
therapy	O
and	O
tolerability	O
was	O
good	O
.	O

First	O
clinical	O
control	O
with	O
second	O
angiography	O
was	O
performed	O
6	O
weeks	O
after	O
irradiation	O
,	O
then	O
in	O
3	O
-	O
month	O
intervals	O
.	O

Eighteen	O
patients	B-Organism
with	O
SRNV	O
refusing	O
radiotherapy	O
served	O
as	O
a	O
control	O
group	O
and	O
were	O
matched	O
with	O
18	O
irradiated	O
patients	B-Organism
.	O

After	O
7	O
months	O
median	O
visual	O
acuity	O
(	O
VA	O
)	O
was	O
20	O
/	O
160	O
for	O
the	O
irradiated	O
and	O
20	O
/	O
400	O
for	O
the	O
untreated	O
patients	B-Organism
.	O

One	O
year	O
after	O
radiotherapy	O
final	O
median	O
VA	O
was	O
20	O
/	O
400	O
in	O
both	O
groups	O
.	O

CONCLUSION	O
:	O
These	O
results	O
suggest	O
that	O
16	O
Gy	O
of	O
conventionally	O
fractionated	O
external	O
beam	O
irradiation	O
slows	O
down	O
the	O
visual	O
loss	O
in	O
exudative	O
AMD	O
for	O
only	O
a	O
few	O
months	O
.	O

Patients	B-Organism
'	O
reading	O
vision	O
could	O
not	O
be	O
saved	O
for	O
a	O
long	O
-	O
term	O
run	O
.	O

Increased	O
serum	B-Organism_substance
levels	O
of	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
in	O
patients	B-Organism
with	O
renal	B-Cancer
cell	I-Cancer
carcinoma	I-Cancer
.	O

Neovascularization	O
,	O
an	O
essential	O
event	O
for	O
the	O
growth	O
of	O
solid	B-Cancer
tumors	I-Cancer
,	O
is	O
regulated	O
by	O
a	O
number	O
of	O
angiogenic	O
factors	O
.	O

One	O
such	O
factor	O
,	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
,	O
is	O
considered	O
to	O
exert	O
a	O
potent	O
angiogenic	O
activity	O
,	O
as	O
indicated	O
by	O
immunohistochemical	O
and	O
molecular	O
evidence	O
.	O

In	O
this	O
study	O
we	O
investigated	O
the	O
serum	B-Organism_substance
VEGF	B-Gene_or_gene_product
level	O
(	O
s	O
-	O
VEGF	B-Gene_or_gene_product
)	O
in	O
patients	B-Organism
with	O
renal	B-Cancer
cell	I-Cancer
carcinoma	I-Cancer
(	O
RCC	B-Cancer
)	O
.	O

s	O
-	O
VEGF	B-Gene_or_gene_product
in	O
peripheral	B-Organism_substance
blood	I-Organism_substance
samples	I-Organism_substance
was	O
analyzed	O
in	O
40	O
RCC	B-Cancer
patients	B-Organism
and	O
40	O
patients	B-Organism
without	O
cancer	B-Cancer
(	O
controls	O
)	O
using	O
a	O
sandwich	O
enzyme	O
-	O
linked	O
immunoassay	O
.	O

In	O
20	O
RCC	B-Cancer
patients	B-Organism
,	O
serum	B-Organism_substance
samples	I-Organism_substance
were	O
obtained	O
separately	O
from	O
the	O
bilateral	B-Multi-tissue_structure
renal	I-Multi-tissue_structure
veins	I-Multi-tissue_structure
.	O

s	O
-	O
VEGF	B-Gene_or_gene_product
was	O
also	O
measured	O
before	O
,	O
4	O
and	O
8	O
weeks	O
after	O
nephrectomy	O
in	O
11	O
patients	B-Organism
.	O

There	O
were	O
significant	O
differences	O
in	O
s	O
-	O
VEGF	B-Gene_or_gene_product
between	O
the	O
RCC	B-Cancer
patients	B-Organism
and	O
the	O
controls	O
(	O
207	O
.	O
3	O
+	O
/	O
-	O
32	O
.	O
9	O
vs	O
.	O
71	O
.	O
5	O
+	O
/	O
-	O
9	O
.	O
1	O
pg	O
/	O
ml	O
,	O
mean	O
+	O
/	O
-	O
SE	O
)	O
(	O
P	O
less	O
than	O
0	O
.	O
005	O
)	O
,	O
between	O
the	O
tumor	B-Cancer
-	O
bearing	O
renal	B-Multi-tissue_structure
veins	I-Multi-tissue_structure
and	O
the	O
contralateral	O
ones	O
(	O
P	O
less	O
than	O
0	O
.	O
01	O
)	O
,	O
between	O
the	O
pre	O
-	O
and	O
post	O
-	O
nephrectomy	O
situations	O
(	O
P	O
less	O
than	O
0	O
.	O
01	O
)	O
and	O
among	O
the	O
various	O
parameters	O
of	O
tumor	B-Cancer
status	O
such	O
as	O
tumor	B-Cancer
extent	O
(	O
P	O
less	O
than	O
0	O
.	O
001	O
)	O
and	O
existence	O
of	O
metastasis	O
(	O
P	O
less	O
than	O
0	O
.	O
001	O
)	O
.	O

s	O
-	O
VEGF	B-Gene_or_gene_product
significantly	O
correlated	O
with	O
the	O
tumor	B-Cancer
volume	O
obtained	O
by	O
a	O
three	O
-	O
dimensional	O
measurement	O
(	O
r	O
=	O
0	O
.	O
802	O
,	O
P	O
less	O
than	O
0	O
.	O
0001	O
)	O
.	O

The	O
sensitivity	O
and	O
specificity	O
of	O
s	O
-	O
VEGF	B-Gene_or_gene_product
at	O
the	O
cut	O
-	O
off	O
level	O
of	O
100	O
pg	O
/	O
ml	O
,	O
as	O
determined	O
by	O
the	O
receiver	O
-	O
operating	O
-	O
characteristics	O
curve	O
,	O
were	O
80	O
.	O
0	O
%	O
and	O
72	O
.	O
5	O
%	O
,	O
respectively	O
.	O

The	O
results	O
indicate	O
that	O
tumor	B-Tissue
tissue	I-Tissue
of	O
RCC	B-Cancer
liberates	O
VEGF	B-Gene_or_gene_product
into	O
the	O
systemic	O
blood	B-Organism_substance
flow	O
and	O
that	O
s	O
-	O
VEGF	B-Gene_or_gene_product
is	O
a	O
possible	O
marker	O
for	O
RCC	B-Cancer
.	O

Insulin	B-Gene_or_gene_product
-	O
induced	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
expression	O
in	O
retina	B-Multi-tissue_structure
.	O

PURPOSE	O
:	O
Clinical	O
studies	O
have	O
demonstrated	O
that	O
intensive	O
insulin	B-Gene_or_gene_product
therapy	O
causes	O
a	O
transient	O
worsening	O
of	O
retinopathy	O
.	O

The	O
mechanisms	O
underlying	O
the	O
initial	O
insulin	B-Gene_or_gene_product
-	O
induced	O
deterioration	O
of	O
retinal	B-Multi-tissue_structure
status	O
in	O
patients	B-Organism
with	O
diabetes	O
remain	O
unknown	O
.	O

Vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
is	O
known	O
to	O
be	O
operative	O
in	O
the	O
pathogenesis	O
of	O
diabetic	O
retinopathy	O
.	O

The	O
current	O
study	O
was	O
conducted	O
to	O
characterize	O
the	O
effect	O
of	O
insulin	B-Gene_or_gene_product
on	O
retinal	B-Multi-tissue_structure
VEGF	B-Gene_or_gene_product
gene	O
expression	O
in	O
vitro	O
and	O
in	O
vivo	O
.	O

METHODS	O
:	O
The	O
effect	O
of	O
insulin	B-Gene_or_gene_product
on	O
VEGF	B-Gene_or_gene_product
expression	O
in	O
vivo	O
was	O
examined	O
by	O
in	O
situ	O
hybridization	O
studies	O
of	O
rat	B-Organism
retinal	B-Multi-tissue_structure
VEGF	B-Gene_or_gene_product
transcripts	O
.	O

To	O
examine	O
the	O
mechanisms	O
by	O
which	O
insulin	B-Gene_or_gene_product
regulates	O
VEGF	B-Gene_or_gene_product
expression	O
,	O
human	B-Organism
retinal	B-Cell
pigment	I-Cell
epithelial	I-Cell
(	I-Cell
RPE	I-Cell
)	I-Cell
cells	I-Cell
were	O
exposed	O
to	O
insulin	B-Gene_or_gene_product
,	O
and	O
VEGF	B-Gene_or_gene_product
mRNA	O
levels	O
were	O
quantified	O
with	O
RNase	B-Gene_or_gene_product
protection	O
assays	O
(	O
RPAs	O
)	O
.	O

Conditioned	O
media	O
from	O
insulin	B-Gene_or_gene_product
-	O
treated	O
RPE	B-Cell
cells	I-Cell
were	O
assayed	O
for	O
VEGF	B-Gene_or_gene_product
protein	O
and	O
capillary	B-Cell
endothelial	I-Cell
cell	I-Cell
proliferation	O
.	O

The	O
capacity	O
of	O
insulin	B-Gene_or_gene_product
to	O
stimulate	O
the	O
VEGF	B-Gene_or_gene_product
promoter	O
linked	O
to	O
a	O
luciferase	B-Gene_or_gene_product
reporter	O
gene	O
was	O
characterized	O
in	O
transient	O
transfection	O
assays	O
.	O

RESULTS	O
:	O
Insulin	B-Gene_or_gene_product
increased	O
VEGF	B-Gene_or_gene_product
mRNA	O
levels	O
in	O
the	O
ganglion	B-Tissue
,	O
inner	B-Tissue
nuclear	I-Tissue
,	O
and	O
RPE	B-Tissue
cell	I-Tissue
layers	I-Tissue
.	O

In	O
vitro	O
,	O
insulin	B-Gene_or_gene_product
increased	O
VEGF	B-Gene_or_gene_product
mRNA	O
levels	O
in	O
human	B-Organism
RPE	B-Cell
cells	I-Cell
and	O
enhanced	O
VEGF	B-Gene_or_gene_product
promoter	O
activity	O
without	O
affecting	O
transcript	O
stability	O
.	O

Insulin	B-Gene_or_gene_product
treatment	O
also	O
increased	O
VEGF	B-Gene_or_gene_product
protein	O
levels	O
in	O
conditioned	O
RPE	B-Cell
cell	I-Cell
media	O
in	O
a	O
dose	O
-	O
dependent	O
manner	O
with	O
a	O
median	O
effective	O
concentration	O
of	O
5	O
nM	O
.	O

The	O
insulin	B-Gene_or_gene_product
-	O
conditioned	O
RPE	B-Cell
cell	I-Cell
media	O
stimulated	O
capillary	B-Cell
endothelial	I-Cell
cell	I-Cell
proliferation	O
,	O
an	O
effect	O
that	O
was	O
completely	O
blocked	O
by	O
anti	O
-	O
VEGF	B-Gene_or_gene_product
neutralizing	O
antibody	O
.	O

CONCLUSIONS	O
:	O
Insulin	B-Gene_or_gene_product
increases	O
VEGF	B-Gene_or_gene_product
mRNA	O
and	O
secreted	O
protein	O
levels	O
in	O
RPE	B-Cell
cells	I-Cell
through	O
enhanced	O
transcription	O
of	O
the	O
VEGF	B-Gene_or_gene_product
gene	O
.	O

Intensive	O
insulin	B-Gene_or_gene_product
therapy	O
may	O
cause	O
a	O
transient	O
worsening	O
of	O
retinopathy	O
in	O
patients	B-Organism
with	O
diabetes	O
through	O
increased	O
retinal	B-Multi-tissue_structure
VEGF	B-Gene_or_gene_product
gene	O
expression	O
.	O

The	O
antiangiogenic	O
agent	O
linomide	B-Simple_chemical
inhibits	O
the	O
growth	O
rate	O
of	O
von	B-Cancer
Hippel	I-Cancer
-	I-Cancer
Lindau	I-Cancer
paraganglioma	I-Cancer
xenografts	I-Cancer
to	O
mice	B-Organism
.	O

The	O
aim	O
of	O
this	O
study	O
was	O
to	O
ascertain	O
the	O
potential	O
usefulness	O
of	O
the	O
antiangiogenic	O
compound	O
linomide	B-Simple_chemical
for	O
treatment	O
of	O
von	B-Cancer
Hippel	I-Cancer
-	I-Cancer
Lindau	I-Cancer
(	I-Cancer
VHL	I-Cancer
)	I-Cancer
-	I-Cancer
related	I-Cancer
tumors	I-Cancer
.	O

Paraganglioma	B-Tissue
tissue	I-Tissue
fragments	I-Tissue
obtained	O
at	O
surgery	O
from	O
a	O
VHL	O
type	O
2a	O
patient	B-Organism
were	O
transplanted	O
s	O
.	O
c	O
.	O
to	O
male	O
BALB	B-Organism
/	I-Organism
c	I-Organism
nu	I-Organism
/	I-Organism
nu	I-Organism
(	I-Organism
nude	I-Organism
)	I-Organism
mice	I-Organism
:	O
(	O
a	O
)	O
2	O
-	O
3	O
-	O
mm	O
fragments	O
for	O
"	O
prevention	O
"	O
experiments	O
;	O
and	O
(	O
b	O
)	O
2	O
-	O
3	O
-	O
mm	O
fragments	O
allowed	O
to	O
grow	O
to	O
1	O
cm	O
for	O
"	O
intervention	O
"	O
studies	O
.	O

Both	O
groups	O
received	O
either	O
0	O
.	O
5	O
mg	O
/	O
ml	O
linomide	B-Simple_chemical
in	O
drinking	O
water	O
or	O
acidified	O
water	O
and	O
were	O
followed	O
until	O
tumor	B-Cancer
diameter	O
reached	O
3	O
cm	O
or	O
for	O
4	O
weeks	O
.	O

In	O
both	O
the	O
prevention	O
and	O
intervention	O
experiments	O
,	O
a	O
significant	O
diminution	O
of	O
tumor	B-Cancer
size	O
and	O
weight	O
was	O
observed	O
in	O
the	O
drug	O
-	O
treated	O
animals	O
.	O

In	O
vivo	O
nuclear	O
magnetic	O
resonance	O
analysis	O
of	O
tumor	B-Cancer
blood	B-Organism_substance
flow	O
in	O
linomide	B-Simple_chemical
-	O
treated	O
animals	O
showed	O
localization	O
of	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
almost	O
exclusively	O
to	O
the	O
periphery	O
of	O
the	O
poorly	O
vascularized	O
tumors	B-Cancer
with	O
a	O
significant	O
reduction	O
of	O
both	O
vascular	B-Multi-tissue_structure
functionality	O
and	O
vasodilation	O
.	O

Histological	O
examination	O
of	O
tumors	B-Cancer
from	O
linomide	B-Simple_chemical
-	O
treated	O
animals	O
revealed	O
marked	O
avascularity	O
.	O

Treated	O
animals	O
also	O
displayed	O
a	O
2	O
.	O
4	O
-	O
fold	O
reduction	O
of	O
tumor	B-Cancer
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
mRNA	O
levels	O
.	O

Taken	O
together	O
,	O
our	O
data	O
indicate	O
that	O
in	O
VHL	O
disease	O
,	O
therapy	O
directed	O
at	O
inhibition	O
of	O
constitutively	O
expressed	O
VEGF	B-Gene_or_gene_product
induction	O
of	O
angiogenesis	O
by	O
VHL	B-Cancer
tumors	I-Cancer
may	O
constitute	O
an	O
effective	O
medical	O
treatment	O
.	O

LMP1	B-Gene_or_gene_product
of	O
Epstein	B-Organism
-	I-Organism
Barr	I-Organism
virus	I-Organism
induces	O
proliferation	O
of	O
primary	O
mouse	B-Organism
embryonic	B-Cell
fibroblasts	I-Cell
and	O
cooperatively	O
transforms	O
the	O
cells	B-Cell
with	O
a	O
p16	B-Gene_or_gene_product
-	O
insensitive	O
CDK4	B-Gene_or_gene_product
oncogene	O
.	O

The	O
latent	O
membrane	O
protein	O
LMP1	B-Gene_or_gene_product
of	O
Epstein	B-Organism
-	I-Organism
Barr	I-Organism
virus	I-Organism
(	O
EBV	B-Organism
)	O
is	O
often	O
present	O
in	O
EBV	B-Organism
-	O
associated	O
malignancies	B-Cancer
including	O
nasopharyngeal	B-Cancer
carcinoma	I-Cancer
and	O
Hodgkin	B-Cancer
'	I-Cancer
s	I-Cancer
lymphoma	I-Cancer
.	O

Previous	O
work	O
demonstrates	O
that	O
the	O
LMP1	B-Gene_or_gene_product
gene	O
of	O
EBV	B-Organism
is	O
sufficient	O
to	O
transform	O
certain	O
established	O
rodent	O
fibroblast	B-Cell
cell	I-Cell
lines	I-Cell
and	O
to	O
induce	O
the	O
tumorigenicity	O
of	O
some	O
human	B-Organism
epithelial	B-Cell
cell	I-Cell
lines	I-Cell
.	O

In	O
addition	O
,	O
LMP1	B-Gene_or_gene_product
plays	O
pleiotropic	O
roles	O
in	O
cell	B-Cell
growth	O
arrest	O
,	O
differentiation	O
,	O
and	O
apoptosis	O
,	O
depending	O
on	O
the	O
background	O
of	O
the	O
target	O
cells	B-Cell
.	O

To	O
examine	O
the	O
roles	O
of	O
LMP1	B-Gene_or_gene_product
in	O
cell	B-Cell
proliferation	O
and	O
growth	O
regulation	O
in	O
primary	B-Cell
culture	I-Cell
cells	I-Cell
,	O
we	O
constructed	O
a	O
recombinant	O
retrovirus	B-Organism
containing	O
an	O
LMP1	B-Gene_or_gene_product
gene	O
.	O

With	O
this	O
retrovirus	B-Organism
,	O
LMP1	B-Gene_or_gene_product
was	O
shown	O
to	O
stimulate	O
the	O
proliferation	O
of	O
primary	O
mouse	B-Organism
embryonic	B-Cell
fibroblasts	I-Cell
(	O
MEF	B-Cell
cells	I-Cell
)	O
.	O

It	O
has	O
a	O
mitogenic	O
activity	O
for	O
MEF	B-Cell
cells	I-Cell
,	O
as	O
demonstrated	O
by	O
an	O
immediate	O
induction	O
of	O
cell	B-Cell
doubling	O
time	O
.	O

In	O
addition	O
,	O
it	O
significantly	O
extends	O
the	O
passage	O
number	O
of	O
MEF	B-Cell
cells	I-Cell
to	O
more	O
than	O
30	O
after	O
retroviral	B-Organism
infection	O
,	O
compared	O
with	O
less	O
than	O
5	O
for	O
uninfected	O
MEF	B-Cell
cells	I-Cell
.	O

Furthermore	O
,	O
LMP1	B-Gene_or_gene_product
cooperates	O
with	O
a	O
p16	B-Gene_or_gene_product
-	O
insensitive	O
CDK4	B-Gene_or_gene_product
(	O
R24C	O
)	O
oncogene	O
in	O
transforming	O
MEF	B-Cell
cells	I-Cell
.	O

Our	O
results	O
provide	O
the	O
first	O
evidence	O
of	O
the	O
abilities	O
of	O
the	O
LMP1	B-Gene_or_gene_product
gene	O
,	O
acting	O
alone	O
,	O
to	O
effectively	O
induce	O
the	O
proliferation	O
of	O
primary	O
MEF	B-Cell
cells	I-Cell
and	O
of	O
its	O
cooperativity	O
with	O
another	O
cellular	B-Cell
oncogene	O
in	O
transforming	O
primary	O
cells	B-Cell
.	O

Autotaxin	B-Gene_or_gene_product
(	O
ATX	B-Gene_or_gene_product
)	O
,	O
a	O
potent	O
tumor	B-Cancer
motogen	O
,	O
augments	O
invasive	O
and	O
metastatic	O
potential	O
of	O
ras	B-Gene_or_gene_product
-	O
transformed	O
cells	B-Cell
.	O

Autotaxin	B-Gene_or_gene_product
(	O
ATX	B-Gene_or_gene_product
)	O
,	O
an	O
exo	O
-	O
nucleotide	O
pyrophosphatase	O
and	O
phosphodiesterase	O
,	O
was	O
originally	O
isolated	O
as	O
a	O
potent	O
stimulator	O
of	O
tumor	B-Cell
cell	I-Cell
motility	O
.	O

In	O
order	O
to	O
study	O
whether	O
ATX	B-Gene_or_gene_product
expression	O
affects	O
motility	O
-	O
dependent	O
processes	O
such	O
as	O
invasion	O
and	O
metastasis	O
,	O
we	O
stably	O
transfected	O
full	O
-	O
length	O
ATX	B-Gene_or_gene_product
cDNA	O
into	O
two	O
non	O
-	O
expressing	O
cell	B-Cell
lines	I-Cell
,	O
parental	O
and	O
ras	B-Gene_or_gene_product
-	O
transformed	O
NIH3T3	B-Cell
(	I-Cell
clone7	I-Cell
)	I-Cell
cells	I-Cell
.	O

The	O
effect	O
of	O
ATX	B-Gene_or_gene_product
secretion	O
on	O
in	O
vitro	O
cell	B-Cell
motility	O
was	O
variable	O
.	O

The	O
ras	B-Gene_or_gene_product
-	O
transformed	O
,	O
ATX	B-Gene_or_gene_product
-	O
secreting	O
subclones	B-Cell
had	O
enhanced	O
motility	O
to	O
ATX	B-Gene_or_gene_product
as	O
chemoattractant	O
,	O
but	O
there	O
was	O
little	O
difference	O
in	O
the	O
motility	O
responses	O
of	O
NIH3T3	B-Cell
cells	I-Cell
transfected	O
with	O
atx	B-Gene_or_gene_product
,	O
an	O
inactive	O
mutant	O
gene	O
,	O
or	O
empty	O
vector	O
.	O

In	O
MatrigelTM	O
invasion	O
assays	O
,	O
all	O
subclones	B-Cell
,	O
which	O
secreted	O
enzymatically	O
active	O
ATX	B-Gene_or_gene_product
,	O
demonstrated	O
greater	O
spontaneous	O
and	O
ATX	B-Gene_or_gene_product
-	O
stimulated	O
invasion	O
than	O
appropriate	O
controls	O
.	O

This	O
difference	O
in	O
invasiveness	O
was	O
not	O
caused	O
by	O
differences	O
in	O
gelatinase	B-Gene_or_gene_product
production	O
,	O
which	O
was	O
constant	O
within	O
each	O
group	O
of	O
transfectants	B-Cell
.	O

In	O
vivo	O
studies	O
with	O
athymic	B-Organism
nude	I-Organism
mice	I-Organism
demonstrated	O
that	O
injection	O
of	O
atx	B-Gene_or_gene_product
-	O
transfected	O
NIH3T3	B-Cell
cells	I-Cell
resulted	O
in	O
a	O
weak	O
tumorigenic	O
capacity	O
with	O
few	O
experimental	O
metastases	O
.	O

Combination	O
of	O
ATX	B-Gene_or_gene_product
expression	O
with	O
ras	B-Gene_or_gene_product
transformation	O
produced	O
cells	B-Cell
with	O
greatly	O
amplified	O
tumorigenesis	O
and	O
metastatic	O
potential	O
compared	O
to	O
ras	B-Gene_or_gene_product
-	O
transformed	O
controls	B-Cell
.	O

Thus	O
,	O
ATX	B-Gene_or_gene_product
appears	O
to	O
augment	O
cellular	B-Cell
characteristics	O
necessary	O
for	O
tumor	B-Cancer
aggressiveness	O
.	O

Oncogenic	O
epidermal	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
mutants	O
with	O
tandem	O
duplication	O
:	O
gene	O
structure	O
and	O
effects	O
on	O
receptor	O
function	O
.	O

A	O
number	O
of	O
epidermal	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
(	O
EGFR	B-Gene_or_gene_product
)	O
deletion	O
mutants	O
have	O
been	O
identified	O
in	O
gliomas	B-Cancer
,	O
in	O
which	O
the	O
EGFR	B-Gene_or_gene_product
gene	O
is	O
frequently	O
amplified	O
and	O
rearranged	O
.	O

We	O
have	O
previously	O
characterized	O
the	O
structure	O
of	O
a	O
gene	O
in	O
A	B-Cell
-	I-Cell
172	I-Cell
human	I-Cell
glioma	I-Cell
cells	I-Cell
that	O
encodes	O
a	O
190	O
-	O
kDa	O
EGFR	B-Gene_or_gene_product
mutant	O
with	O
tandem	O
duplication	O
of	O
the	O
tyrosine	O
kinase	O
(	O
TK	O
)	O
and	O
calcium	B-Simple_chemical
-	O
mediated	O
internalization	O
(	O
CAIN	O
)	O
domains	O
.	O

Here	O
we	O
describe	O
a	O
185	O
-	O
kDa	O
tandem	O
duplication	O
mutant	O
(	O
TDM	O
)	O
that	O
is	O
expressed	O
in	O
KE	B-Cell
and	O
A	B-Cell
-	I-Cell
1235	I-Cell
glioma	I-Cell
cells	I-Cell
,	O
along	O
with	O
certain	O
functional	O
characteristics	O
of	O
the	O
mutants	O
.	O

The	O
corresponding	O
transcripts	O
in	O
KE	B-Cell
and	O
A	B-Cell
-	I-Cell
1235	I-Cell
cells	I-Cell
contain	O
1053	O
additional	O
nucleotides	O
representing	O
an	O
in	O
-	O
frame	O
duplication	O
of	O
exons	O
18	O
through	O
25	O
which	O
encode	O
the	O
entire	O
TK	O
region	O
and	O
a	O
portion	O
of	O
the	O
CAIN	O
domain	O
.	O

As	O
with	O
duplication	O
of	O
the	O
entire	O
TK	O
/	O
CAIN	O
region	O
(	O
exons	O
18	O
-	O
26	O
)	O
in	O
A	B-Cell
-	I-Cell
172	I-Cell
cells	I-Cell
,	O
duplication	O
of	O
exons	O
18	O
-	O
25	O
is	O
associated	O
with	O
a	O
specific	O
genomic	O
rearrangement	O
between	O
flanking	O
introns	O
.	O

Involved	O
introns	O
contain	O
homology	O
to	O
recombination	O
signal	O
sequence	O
(	O
RSS	O
)	O
heptamers	O
present	O
in	O
the	O
V	O
(	O
D	O
)	O
J	O
region	O
of	O
the	O
T	B-Gene_or_gene_product
lymphocyte	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
gene	O
.	O

In	O
defined	O
medium	O
,	O
both	O
oncogenic	O
TDM	O
are	O
constitutively	O
autophosphorylated	O
and	O
inefficiently	O
downregulated	O
.	O

High	O
-	O
affinity	O
binding	O
is	O
reduced	O
in	O
EGFR	B-Gene_or_gene_product
.	O
TDM	O
/	O
18	O
-	O
26	O
,	O
although	O
the	O
t1	O
/	O
2	O
of	O
receptor	O
internalization	O
is	O
not	O
prolonged	O
.	O

Ciprofloxacin	B-Simple_chemical
mediated	O
cell	B-Cell
growth	O
inhibition	O
,	O
S	O
/	O
G2	O
-	O
M	O
cell	B-Cell
cycle	O
arrest	O
,	O
and	O
apoptosis	O
in	O
a	O
human	B-Organism
transitional	B-Cancer
cell	I-Cancer
carcinoma	I-Cancer
of	O
the	O
bladder	B-Cell
cell	I-Cell
line	I-Cell
.	O

The	O
second	O
most	O
prevalent	O
urological	B-Cancer
malignancy	I-Cancer
in	O
middle	O
aged	O
and	O
elderly	O
men	B-Organism
is	O
bladder	B-Cancer
cancer	I-Cancer
,	O
with	O
90	O
%	O
of	O
the	O
cases	O
being	O
transitional	B-Cancer
cell	I-Cancer
carcinomas	I-Cancer
.	O

The	O
success	O
of	O
current	O
systemic	O
and	O
intravesical	O
therapeutic	O
agents	O
,	O
such	O
as	O
cisplatin	B-Simple_chemical
,	O
thiotepa	B-Simple_chemical
,	O
Adriamycin	B-Simple_chemical
,	O
mitomycin	B-Simple_chemical
C	I-Simple_chemical
,	O
and	O
bacillus	B-Organism
Calmette	I-Organism
-	I-Organism
Guerin	I-Organism
,	O
is	O
limited	O
with	O
recurrence	O
rates	O
reduced	O
to	O
17	O
-	O
44	O
%	O
.	O

In	O
addition	O
,	O
most	O
of	O
these	O
agents	O
require	O
instrumentation	O
of	O
the	O
urinary	B-Organism_subdivision
tract	I-Organism_subdivision
and	O
are	O
delivered	O
at	O
a	O
significant	O
cost	O
and	O
potential	O
morbidity	O
to	O
the	O
patient	B-Organism
.	O

Fluroquinolone	B-Simple_chemical
antibiotics	O
such	O
as	O
ciprofloxacin	B-Simple_chemical
,	O
which	O
can	O
be	O
administered	O
p	O
.	O
o	O
.	O
,	O
may	O
have	O
a	O
profound	O
effect	O
in	O
bladder	B-Cancer
cancer	I-Cancer
management	O
.	O

This	O
is	O
primarily	O
based	O
on	O
limited	O
in	O
vitro	O
studies	O
on	O
tumor	B-Cell
cells	I-Cell
derived	O
from	O
transitional	B-Cancer
cell	I-Cancer
carcinoma	I-Cancer
of	O
the	O
bladder	B-Organ
that	O
revealed	O
a	O
dose	O
-	O
and	O
time	O
-	O
dependent	O
inhibition	O
of	O
cell	B-Cell
growth	O
by	O
ciprofloxacin	B-Simple_chemical
at	O
concentrations	O
that	O
are	O
easily	O
attainable	O
in	O
the	O
urine	B-Organism_substance
of	O
patients	B-Organism
.	O

However	O
,	O
the	O
mechanism	O
(	O
s	O
)	O
by	O
which	O
ciprofloxacin	B-Simple_chemical
elicits	O
its	O
biological	O
effects	O
on	O
bladder	B-Cell
cancer	I-Cell
cells	I-Cell
is	O
not	O
well	O
documented	O
.	O

Our	O
experimental	O
data	O
confirm	O
previous	O
studies	O
showing	O
the	O
in	O
vitro	O
cell	B-Cell
growth	O
inhibition	O
of	O
the	O
transitional	B-Cancer
cell	I-Cancer
carcinoma	I-Cancer
of	O
the	O
bladder	B-Cell
cell	I-Cell
line	I-Cell
HTB9	I-Cell
and	O
further	O
showed	O
the	O
induction	O
of	O
cell	B-Cell
cycle	O
arrest	O
at	O
the	O
S	O
/	O
G2	O
-	O
M	O
checkpoints	O
.	O

In	O
addition	O
,	O
we	O
found	O
down	O
-	O
regulation	O
of	O
cyclin	B-Gene_or_gene_product
B	I-Gene_or_gene_product
,	O
cyclin	B-Gene_or_gene_product
E	I-Gene_or_gene_product
,	O
and	O
dephosphorylation	O
of	O
cdk2	B-Gene_or_gene_product
in	O
ciprofloxacin	B-Simple_chemical
-	O
treated	O
bladder	B-Cell
tumor	I-Cell
cells	I-Cell
.	O

There	O
was	O
also	O
an	O
up	O
-	O
regulation	O
of	O
Bax	B-Gene_or_gene_product
,	O
which	O
altered	O
the	O
Bax	B-Gene_or_gene_product
:	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
ratio	O
,	O
which	O
may	O
be	O
responsible	O
for	O
mitochondrial	B-Cellular_component
depolarization	O
reported	O
to	O
be	O
involved	O
prior	O
to	O
the	O
induction	O
of	O
apoptosis	O
.	O

The	O
cyclin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
dependent	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
inhibitor	O
p21WAF1	B-Gene_or_gene_product
level	O
was	O
found	O
to	O
be	O
decreased	O
within	O
12	O
h	O
of	O
ciprofloxacin	B-Simple_chemical
treatment	O
and	O
disappeared	O
completely	O
when	O
HTB9	B-Cell
cells	I-Cell
were	O
treated	O
with	O
200	O
microg	O
/	O
ml	O
ciprofloxacin	B-Simple_chemical
for	O
24	O
h	O
.	O

The	O
down	O
-	O
regulation	O
of	O
p21WAF1	B-Gene_or_gene_product
closely	O
correlated	O
with	O
poly	B-Gene_or_gene_product
(	I-Gene_or_gene_product
ADP	I-Gene_or_gene_product
-	I-Gene_or_gene_product
ribose	I-Gene_or_gene_product
)	I-Gene_or_gene_product
polymerase	I-Gene_or_gene_product
cleavage	O
and	O
CPP32	B-Gene_or_gene_product
activation	O
.	O

Recent	O
studies	O
revealed	O
that	O
p21WAF1	B-Gene_or_gene_product
protects	O
cells	B-Cell
from	O
apoptosis	O
by	O
arresting	O
them	O
in	O
G1	O
and	O
further	O
binds	O
to	O
pro	B-Gene_or_gene_product
-	I-Gene_or_gene_product
caspase	I-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
,	O
preventing	O
its	O
activation	O
and	O
thus	O
,	O
inhibiting	O
the	O
apoptotic	O
cascade	O
.	O

Hence	O
,	O
the	O
down	O
-	O
regulation	O
of	O
p21WAF1	B-Gene_or_gene_product
,	O
together	O
with	O
the	O
alterations	O
in	O
Bax	B-Gene_or_gene_product
and	O
cdk2	B-Gene_or_gene_product
as	O
observed	O
in	O
our	O
studies	O
,	O
may	O
define	O
a	O
novel	O
mechanism	O
by	O
which	O
ciprofloxacin	B-Simple_chemical
inhibits	O
tumor	B-Cell
cell	I-Cell
growth	O
and	O
induces	O
apoptotic	O
cell	B-Cell
death	O
.	O

The	O
results	O
of	O
our	O
current	O
studies	O
provide	O
strong	O
experimental	O
evidence	O
for	O
the	O
use	O
of	O
ciprofloxacin	B-Simple_chemical
as	O
a	O
potential	O
preventive	O
and	O
/	O
or	O
therapeutic	O
agent	O
for	O
the	O
management	O
of	O
transitional	B-Cancer
cell	I-Cancer
carcinoma	I-Cancer
of	O
the	O
bladder	B-Organ
.	O

Vascular	B-Multi-tissue_structure
proliferation	O
and	O
enhanced	O
expression	O
of	O
endothelial	B-Gene_or_gene_product
nitric	I-Gene_or_gene_product
oxide	I-Gene_or_gene_product
synthase	I-Gene_or_gene_product
in	O
human	B-Organism
peritoneum	B-Multi-tissue_structure
exposed	O
to	O
long	O
-	O
term	O
peritoneal	B-Multi-tissue_structure
dialysis	O
.	O

Long	O
-	O
term	O
peritoneal	B-Multi-tissue_structure
dialysis	O
(	O
PD	O
)	O
is	O
associated	O
with	O
alterations	O
in	O
peritoneal	B-Multi-tissue_structure
permeability	O
and	O
loss	O
of	O
ultrafiltration	O
.	O

These	O
changes	O
originate	O
from	O
increased	O
peritoneal	B-Immaterial_anatomical_entity
surface	I-Immaterial_anatomical_entity
area	I-Immaterial_anatomical_entity
,	O
but	O
the	O
morphologic	O
and	O
molecular	O
mechanisms	O
involved	O
remain	O
unknown	O
.	O

The	O
hypothesis	O
that	O
modifications	O
of	O
activity	O
and	O
/	O
or	O
expression	O
of	O
nitric	B-Gene_or_gene_product
oxide	I-Gene_or_gene_product
synthase	I-Gene_or_gene_product
(	O
NOS	B-Gene_or_gene_product
)	O
isozymes	O
might	O
play	O
a	O
role	O
in	O
these	O
modifications	O
,	O
via	O
enhanced	O
local	O
production	O
of	O
nitric	B-Simple_chemical
oxide	I-Simple_chemical
,	O
was	O
tested	O
in	O
this	O
study	O
.	O

NOS	B-Gene_or_gene_product
activities	O
were	O
measured	O
by	O
the	O
L	O
-	O
citrulline	O
assay	O
in	O
peritoneal	B-Multi-tissue_structure
biopsies	I-Multi-tissue_structure
from	O
seven	O
control	O
subjects	O
,	O
eight	O
uremic	O
patients	B-Organism
immediately	O
before	O
the	O
onset	O
of	O
PD	O
,	O
and	O
13	O
uremic	O
patients	B-Organism
on	O
short	O
-	O
term	O
(	O
less	O
than	O
18	O
mo	O
,	O
n	O
=	O
6	O
)	O
or	O
long	O
-	O
term	O
(	O
greater	O
than	O
18	O
mo	O
,	O
n	O
=	O
7	O
)	O
PD	O
.	O

Peritoneal	B-Multi-tissue_structure
NOS	B-Gene_or_gene_product
activity	O
is	O
increased	O
fivefold	O
in	O
long	O
-	O
term	O
PD	O
patients	B-Organism
compared	O
with	O
control	O
subjects	O
.	O

In	O
uremic	O
patients	B-Organism
,	O
NOS	B-Gene_or_gene_product
activity	O
is	O
positively	O
correlated	O
with	O
the	O
duration	O
of	O
PD	O
.	O

Increased	O
NOS	B-Gene_or_gene_product
activity	O
is	O
mediated	O
solely	O
by	O
Ca	O
(	O
2	O
+	O
)	O
-	O
dependent	O
NOS	B-Gene_or_gene_product
and	O
,	O
as	O
shown	O
by	O
immunoblotting	O
,	O
an	O
upregulation	O
of	O
endothelial	B-Gene_or_gene_product
NOS	I-Gene_or_gene_product
.	O

The	O
biologic	O
relevance	O
of	O
increased	O
NOS	B-Gene_or_gene_product
in	O
long	O
-	O
term	O
PD	O
was	O
demonstrated	O
by	O
enhanced	O
nitrotyrosine	B-Simple_chemical
immunoreactivity	O
and	O
a	O
significant	O
increase	O
in	O
vascular	B-Multi-tissue_structure
density	O
and	O
endothelial	B-Tissue
area	I-Tissue
in	O
the	O
peritoneum	B-Multi-tissue_structure
.	O

Immunoblotting	O
and	O
immunostaining	O
studies	O
demonstrated	O
an	O
upregulation	O
of	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
mostly	O
along	O
the	O
endothelium	B-Tissue
lining	O
peritoneal	B-Multi-tissue_structure
blood	I-Multi-tissue_structure
vessels	I-Multi-tissue_structure
in	O
long	O
-	O
term	O
PD	O
patients	B-Organism
.	O

In	O
the	O
latter	O
,	O
VEGF	B-Gene_or_gene_product
colocalized	O
with	O
the	O
advanced	O
glycation	O
end	O
product	O
pentosidine	B-Simple_chemical
deposits	O
.	O

These	O
data	O
provide	O
a	O
morphologic	O
(	O
angiogenesis	O
and	O
increased	O
endothelial	B-Tissue
area	I-Tissue
)	O
and	O
molecular	O
(	O
enhanced	O
NOS	B-Gene_or_gene_product
activity	O
and	O
endothelial	B-Gene_or_gene_product
NOS	I-Gene_or_gene_product
upregulation	O
)	O
basis	O
for	O
explaining	O
the	O
permeability	O
changes	O
observed	O
in	O
long	O
-	O
term	O
PD	O
.	O

They	O
also	O
support	O
the	O
implication	O
of	O
local	O
advanced	O
glycation	O
end	O
product	O
deposits	O
and	O
liberation	O
of	O
VEGF	B-Gene_or_gene_product
in	O
that	O
process	O
.	O

Active	O
hair	B-Multi-tissue_structure
growth	O
(	O
anagen	O
)	O
is	O
associated	O
with	O
angiogenesis	O
.	O

After	O
the	O
completion	O
of	O
skin	B-Organ
development	O
,	O
angiogenesis	O
,	O
i	O
.	O
e	O
.	O
,	O
the	O
growth	O
of	O
new	O
capillaries	B-Tissue
from	O
pre	O
-	O
existing	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
,	O
is	O
held	O
to	O
occur	O
in	O
the	O
skin	B-Organ
only	O
under	O
pathologic	O
conditions	O
.	O

It	O
has	O
long	O
been	O
noted	O
,	O
however	O
,	O
that	O
hair	B-Multi-tissue_structure
follicle	I-Multi-tissue_structure
cycling	O
is	O
associated	O
with	O
prominent	O
changes	O
in	O
skin	B-Organ
perfusion	O
,	O
that	O
the	O
epithelial	B-Tissue
hair	I-Tissue
bulbs	I-Tissue
of	O
anagen	B-Multi-tissue_structure
follicles	I-Multi-tissue_structure
display	O
angiogenic	O
properties	O
,	O
and	O
that	O
the	O
follicular	B-Tissue
dermal	I-Tissue
papilla	I-Tissue
can	O
produce	O
angiogenic	O
factors	O
.	O

Despite	O
these	O
suggestive	O
observations	O
,	O
no	O
formal	O
proof	O
is	O
as	O
yet	O
available	O
for	O
the	O
concept	O
that	O
angiogenesis	O
is	O
a	O
physiologic	O
event	O
that	O
occurs	O
all	O
over	O
the	O
mature	O
mammalian	O
integument	O
whenever	O
hair	B-Multi-tissue_structure
follicles	I-Multi-tissue_structure
switch	O
from	O
resting	O
(	O
telogen	O
)	O
to	O
active	O
growth	O
(	O
anagen	O
)	O
.	O

This	O
study	O
uses	O
quantitative	O
histomorphometry	O
and	O
double	O
-	O
immunohistologic	O
detection	O
techniques	O
for	O
the	O
demarcation	O
of	O
proliferating	O
endothelial	B-Cell
cells	I-Cell
,	O
to	O
show	O
that	O
synchronized	O
hair	B-Multi-tissue_structure
follicle	I-Multi-tissue_structure
cycling	O
in	O
adolescent	O
C57BL	B-Organism
/	I-Organism
6	I-Organism
mice	I-Organism
is	O
associated	O
with	O
substantial	O
angiogenesis	O
,	O
and	O
that	O
inhibiting	O
angiogenesis	O
in	O
vivo	O
by	O
the	O
intraperitoneal	B-Immaterial_anatomical_entity
application	O
of	O
a	O
fumagillin	B-Simple_chemical
derivative	I-Simple_chemical
retards	O
experimentally	O
induced	O
anagen	B-Multi-tissue_structure
development	O
in	O
these	O
mice	B-Organism
.	O

Thus	O
,	O
angiogenesis	O
is	O
a	O
physiologic	O
event	O
in	O
normal	O
postnatal	O
murine	B-Organism
skin	B-Organ
,	O
apparently	O
is	O
dictated	O
by	O
the	O
hair	B-Multi-tissue_structure
follicle	I-Multi-tissue_structure
,	O
and	O
appears	O
to	O
be	O
required	O
for	O
normal	O
anagen	O
development	O
.	O

Anagen	O
-	O
associated	O
angiogenesis	O
offers	O
an	O
attractive	O
model	O
for	O
identifying	O
the	O
physiologic	O
controls	O
of	O
cutaneous	B-Organism_subdivision
angiogenesis	O
,	O
and	O
an	O
interesting	O
system	O
for	O
screening	O
the	O
effects	O
of	O
potential	O
antiangiogenic	O
drugs	O
in	O
vivo	O
.	O

Neuroblastoma	B-Cancer
and	O
hepatocyte	B-Cell
coculture	O
conditioned	O
media	O
alter	O
apoptosis	O
.	O

BACKGROUND	O
:	O
Neuroblastoma	B-Cancer
is	O
a	O
childhood	O
tumor	B-Cancer
that	O
often	O
displays	O
unusual	O
biological	O
behavior	O
.	O

The	O
tumor	B-Cancer
may	O
present	O
with	O
widespread	O
metastases	O
that	O
are	O
unresponsive	O
to	O
aggressive	O
treatment	O
.	O

At	O
other	O
times	O
,	O
both	O
the	O
metastases	O
and	O
the	O
primary	O
tumor	B-Cancer
may	O
spontaneously	O
regress	O
without	O
treatment	O
.	O

Apoptosis	O
,	O
or	O
programmed	O
cell	B-Cell
death	O
,	O
is	O
thought	O
to	O
play	O
a	O
role	O
in	O
the	O
dichotomous	O
behavior	O
of	O
neuroblastoma	B-Cancer
.	O

We	O
hypothesize	O
that	O
neuroblastoma	B-Cell
cells	I-Cell
will	O
interact	O
with	O
host	O
tissues	B-Tissue
to	O
release	O
mediators	O
that	O
affect	O
apoptosis	O
.	O

MATERIALS	O
AND	O
METHODS	O
:	O
Human	B-Organism
neuroblastoma	B-Cell
cells	I-Cell
and	O
human	B-Organism
Chang	B-Cell
hepatocytes	I-Cell
are	O
grown	O
in	O
a	O
noncontact	O
,	O
coculture	O
system	O
.	O

After	O
incubation	O
for	O
4	O
days	O
,	O
the	O
medium	O
from	O
the	O
coculture	O
system	O
is	O
collected	O
.	O

Neuroblastoma	B-Cell
cells	I-Cell
and	O
Chang	B-Cell
hepatocytes	I-Cell
are	O
then	O
plated	O
separately	O
with	O
the	O
conditioned	O
medium	O
and	O
their	O
own	O
standard	O
growth	O
medium	O
as	O
controls	O
.	O

After	O
4	O
days	O
,	O
these	O
cells	B-Cell
are	O
harvested	O
and	O
cytospins	O
made	O
for	O
immunostaining	O
.	O

Tumor	B-Gene_or_gene_product
necrosis	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
(	O
TNF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
)	O
,	O
Fas	B-Gene_or_gene_product
ligand	I-Gene_or_gene_product
,	O
and	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
are	O
measured	O
with	O
immunohistochemistry	O
.	O

Apoptosis	O
is	O
detected	O
with	O
the	O
TUNEL	O
method	O
.	O

Immunostaining	O
data	O
are	O
interpreted	O
with	O
computer	O
image	O
analysis	O
and	O
reported	O
as	O
stain	O
index	O
.	O

TUNEL	O
data	O
are	O
reported	O
as	O
percentage	O
apoptotic	O
cells	B-Cell
.	O

All	O
data	O
are	O
reported	O
as	O
means	O
+	O
/	O
-	O
SEM	O
.	O

Statistical	O
analysis	O
is	O
performed	O
and	O
P	O
<	O
0	O
.	O
05	O
considered	O
significant	O
.	O

RESULTS	O
:	O
Chang	B-Cell
hepatocytes	I-Cell
grown	O
in	O
the	O
coculture	O
conditioned	O
media	O
have	O
an	O
increase	O
in	O
TNF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
and	O
Fas	B-Gene_or_gene_product
ligand	I-Gene_or_gene_product
.	O

The	O
neuroblastoma	B-Cell
cells	I-Cell
have	O
a	O
significant	O
decrease	O
in	O
Fas	B-Gene_or_gene_product
ligand	I-Gene_or_gene_product
.	O

There	O
is	O
a	O
significant	O
increase	O
in	O
the	O
number	O
of	O
apoptotic	O
hepatocytes	B-Cell
when	O
they	O
are	O
cultured	O
in	O
the	O
conditioned	O
media	O
.	O

In	O
contrast	O
,	O
the	O
neuroblastoma	B-Cell
cells	I-Cell
grown	O
in	O
the	O
coculture	O
conditioned	O
media	O
show	O
no	O
increase	O
in	O
apoptosis	O
.	O

Finally	O
,	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
is	O
significantly	O
increased	O
in	O
the	O
neuroblastoma	B-Cell
cells	I-Cell
cultured	O
in	O
the	O
conditioned	O
media	O
.	O

CONCLUSIONS	O
:	O
Neuroblastoma	B-Cell
cells	I-Cell
grown	O
in	O
coculture	O
conditioned	O
media	O
show	O
increased	O
expression	O
of	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
decreased	O
Fas	B-Gene_or_gene_product
ligand	I-Gene_or_gene_product
levels	O
.	O

These	O
changes	O
should	O
diminish	O
apoptosis	O
activity	O
in	O
the	O
tumor	B-Cell
cells	I-Cell
.	O

In	O
contrast	O
,	O
the	O
conditioned	O
media	O
induce	O
elevated	O
levels	O
of	O
proapoptotic	O
mediators	O
in	O
the	O
Chang	O
hepatocytes	B-Cell
.	O

A	O
tumor	B-Cancer
'	O
s	O
ability	O
to	O
successfully	O
metastasize	O
may	O
be	O
dependent	O
on	O
mediators	O
generated	O
in	O
the	O
tumor	B-Cancer
-	O
host	O
interaction	O
,	O
and	O
may	O
not	O
be	O
just	O
an	O
independent	O
characteristic	O
of	O
the	O
tumor	B-Cancer
itself	O
.	O

Neoplastic	O
transformation	O
by	O
Notch	B-Gene_or_gene_product
requires	O
nuclear	B-Cellular_component
localization	O
.	O

Notch	B-Gene_or_gene_product
proteins	O
are	O
plasma	B-Cellular_component
membrane	I-Cellular_component
-	O
spanning	O
receptors	O
that	O
mediate	O
important	O
cell	B-Cell
fate	O
decisions	O
such	O
as	O
differentiation	O
,	O
proliferation	O
,	O
and	O
apoptosis	O
.	O

The	O
mechanism	O
of	O
Notch	B-Gene_or_gene_product
signaling	O
remains	O
poorly	O
understood	O
.	O

However	O
,	O
it	O
is	O
clear	O
that	O
the	O
Notch	B-Gene_or_gene_product
signaling	O
pathway	O
mediates	O
its	O
effects	O
through	O
intercellular	O
contact	O
between	O
neighboring	O
cells	B-Cell
.	O

The	O
prevailing	O
model	O
for	O
Notch	B-Gene_or_gene_product
signaling	O
suggests	O
that	O
ligand	O
,	O
presented	O
on	O
a	O
neighboring	O
cell	B-Cell
,	O
triggers	O
proteolytic	O
processing	O
of	O
Notch	B-Gene_or_gene_product
.	O

Following	O
proteolysis	O
,	O
it	O
is	O
thought	O
that	O
the	O
intracellular	B-Immaterial_anatomical_entity
portion	O
of	O
Notch	B-Gene_or_gene_product
(	O
N	B-Gene_or_gene_product
(	I-Gene_or_gene_product
ic	I-Gene_or_gene_product
)	I-Gene_or_gene_product
)	O
translocates	O
to	O
the	O
nucleus	B-Cellular_component
,	O
where	O
it	O
is	O
involved	O
in	O
regulating	O
gene	O
expression	O
.	O

There	O
is	O
considerable	O
debate	O
concerning	O
where	O
in	O
the	O
cell	B-Cell
Notch	B-Gene_or_gene_product
functions	O
and	O
what	O
proteins	O
serve	O
as	O
effectors	O
of	O
the	O
Notch	B-Gene_or_gene_product
signal	O
.	O

Several	O
Notch	B-Gene_or_gene_product
genes	O
have	O
clearly	O
been	O
shown	O
to	O
be	O
proto	O
-	O
oncogenes	O
in	O
mammalian	B-Cell
cells	I-Cell
.	O

Activation	O
of	O
Notch	B-Gene_or_gene_product
proto	O
-	O
oncogenes	O
has	O
been	O
associated	O
with	O
tumorigenesis	O
in	O
several	O
human	B-Organism
and	O
other	O
mammalian	B-Cancer
cancers	I-Cancer
.	O

Transforming	O
alleles	O
of	O
Notch	B-Gene_or_gene_product
direct	O
the	O
expression	O
of	O
truncated	O
proteins	O
that	O
primarily	O
consist	O
of	O
N	B-Gene_or_gene_product
(	I-Gene_or_gene_product
ic	I-Gene_or_gene_product
)	I-Gene_or_gene_product
and	O
are	O
not	O
tethered	O
to	O
the	O
plasma	B-Cellular_component
membrane	I-Cellular_component
.	O

However	O
,	O
the	O
mechanism	O
by	O
which	O
Notch	B-Gene_or_gene_product
oncoproteins	O
(	O
generically	O
termed	O
here	O
as	O
N	B-Gene_or_gene_product
(	I-Gene_or_gene_product
ic	I-Gene_or_gene_product
)	I-Gene_or_gene_product
)	O
induce	O
neoplastic	O
transformation	O
is	O
not	O
known	O
.	O

Previously	O
we	O
demonstrated	O
that	O
N1	B-Gene_or_gene_product
(	I-Gene_or_gene_product
ic	I-Gene_or_gene_product
)	I-Gene_or_gene_product
and	O
N2	B-Gene_or_gene_product
(	I-Gene_or_gene_product
ic	I-Gene_or_gene_product
)	I-Gene_or_gene_product
could	O
transform	O
E1A	B-Gene_or_gene_product
immortalized	O
baby	O
rat	B-Organism
kidney	B-Cell
cells	I-Cell
(	O
RKE	B-Cell
)	O
in	O
vitro	O
.	O

We	O
now	O
report	O
direct	O
evidence	O
that	O
N1	B-Gene_or_gene_product
(	I-Gene_or_gene_product
ic	I-Gene_or_gene_product
)	I-Gene_or_gene_product
must	O
accumulate	O
in	O
the	O
nucleus	B-Cellular_component
to	O
induce	O
transformation	O
of	O
RKE	B-Cell
cells	I-Cell
.	O

In	O
addition	O
,	O
we	O
define	O
the	O
minimal	O
domain	O
of	O
N1	B-Gene_or_gene_product
(	I-Gene_or_gene_product
ic	I-Gene_or_gene_product
)	I-Gene_or_gene_product
required	O
to	O
induce	O
transformation	O
and	O
present	O
evidence	O
that	O
transformation	O
of	O
RKE	B-Cell
cells	I-Cell
by	O
N1	B-Gene_or_gene_product
(	I-Gene_or_gene_product
ic	I-Gene_or_gene_product
)	I-Gene_or_gene_product
is	O
likely	O
to	O
be	O
through	O
a	O
CBF1	B-Gene_or_gene_product
-	O
independent	O
pathway	O
.	O

Alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
melanocyte	I-Gene_or_gene_product
-	I-Gene_or_gene_product
stimulating	I-Gene_or_gene_product
hormone	I-Gene_or_gene_product
modulates	O
activation	O
of	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappa	I-Gene_or_gene_product
B	I-Gene_or_gene_product
and	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
secretion	O
of	O
interleukin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
in	O
human	B-Organism
dermal	B-Cell
fibroblasts	I-Cell
.	O

Alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
melanocyte	I-Gene_or_gene_product
-	I-Gene_or_gene_product
stimulating	I-Gene_or_gene_product
hormone	I-Gene_or_gene_product
(	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
MSH	I-Gene_or_gene_product
)	O
has	O
evolved	O
as	O
a	O
mediator	O
of	O
diverse	O
biological	O
activities	O
in	O
an	O
ever	O
-	O
growing	O
number	O
of	O
non	B-Cell
-	I-Cell
melanocytic	I-Cell
cell	I-Cell
types	O
.	O

One	O
mechanism	O
by	O
which	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
MSH	I-Gene_or_gene_product
exerts	O
its	O
effects	O
is	O
modulation	O
of	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappa	I-Gene_or_gene_product
B	I-Gene_or_gene_product
.	O

These	O
two	O
transcription	O
factors	O
also	O
play	O
an	O
important	O
role	O
in	O
fibroblasts	B-Cell
,	O
in	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
composition	O
,	O
and	O
in	O
cytokine	O
expression	O
.	O

By	O
use	O
of	O
electric	O
mobility	O
shift	O
assays	O
,	O
we	O
demonstrate	O
that	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
MSH	I-Gene_or_gene_product
(	O
10	O
(	O
-	O
6	O
)	O
to	O
10	O
(	O
-	O
14	O
)	O
M	O
)	O
activates	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
in	O
human	B-Organism
dermal	B-Cell
fibroblasts	I-Cell
,	O
whereas	O
coincubation	O
with	O
interleukin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
(	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
)	O
results	O
in	O
suppression	O
of	O
its	O
activation	O
.	O

alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
MSH	I-Gene_or_gene_product
also	O
induces	O
activation	O
of	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappa	I-Gene_or_gene_product
B	I-Gene_or_gene_product
but	O
does	O
not	O
modulate	O
DNA	B-Cellular_component
binding	O
on	O
costimulation	O
with	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
.	O

Since	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappa	I-Gene_or_gene_product
B	I-Gene_or_gene_product
are	O
key	O
elements	O
in	O
controlling	O
interleukin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
(	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
)	O
transcription	O
,	O
human	B-Organism
fibroblasts	B-Cell
were	O
treated	O
with	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
MSH	I-Gene_or_gene_product
and	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
for	O
24	O
hours	O
,	O
and	O
cytokine	O
levels	O
in	O
the	O
supernatants	O
were	O
measured	O
by	O
ELISA	O
.	O

alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
MSH	I-Gene_or_gene_product
alone	O
had	O
little	O
effect	O
,	O
whereas	O
coincubation	O
with	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
led	O
to	O
marked	O
downregulation	O
of	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
secretion	O
(	O
at	O
most	O
288	O
+	O
/	O
-	O
152	O
ng	O
/	O
mL	O
)	O
when	O
compared	O
to	O
treatment	O
with	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
alone	O
(	O
919	O
+	O
/	O
-	O
157	O
ng	O
/	O
mL	O
)	O
.	O

Our	O
results	O
indicate	O
that	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
MSH	I-Gene_or_gene_product
exerts	O
modulatory	O
effects	O
on	O
the	O
activation	O
of	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappa	I-Gene_or_gene_product
B	I-Gene_or_gene_product
and	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
and	O
that	O
it	O
can	O
regulate	O
chemokine	O
secretion	O
in	O
human	B-Organism
dermal	B-Cell
fibroblasts	I-Cell
.	O

These	O
effects	O
of	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
MSH	I-Gene_or_gene_product
may	O
have	O
important	O
regulatory	O
functions	O
in	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
composition	O
,	O
wound	B-Pathological_formation
healing	O
,	O
or	O
angiogenesis	O
.	O

CD40	B-Gene_or_gene_product
expression	O
on	O
human	B-Organism
lung	B-Cancer
cancer	I-Cancer
correlates	O
with	O
metastatic	O
spread	O
.	O

PURPOSE	O
:	O
The	O
poor	O
prognosis	O
associated	O
with	O
lung	B-Cancer
cancer	I-Cancer
is	O
related	O
to	O
the	O
high	O
incidence	O
of	O
regional	O
and	O
distant	O
metastasis	O
.	O

There	O
is	O
a	O
crucial	O
need	O
to	O
identify	O
parameters	O
that	O
can	O
predict	O
a	O
tendancy	O
to	O
metastatic	O
spread	O
to	O
allow	O
better	O
prognostic	O
evaluation	O
and	O
therapeutic	O
approach	O
.	O

METHODS	O
:	O
Using	O
flow	O
cytometry	O
we	O
evaluated	O
18	O
human	B-Organism
lung	B-Cell
cancer	I-Cell
cell	I-Cell
lines	I-Cell
for	O
the	O
expression	O
of	O
different	O
surface	O
markers	O
on	O
lung	B-Cancer
cancers	I-Cancer
suggested	O
to	O
be	O
possible	O
prognostic	O
parameters	O
,	O
including	O
epidermal	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
(	O
EGFR	B-Gene_or_gene_product
)	O
,	O
intercellular	B-Gene_or_gene_product
adhesion	I-Gene_or_gene_product
molecule	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
ICAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
,	O
Fas	B-Gene_or_gene_product
and	O
CD40	B-Gene_or_gene_product
.	O

RESULTS	O
:	O
No	O
correlation	O
was	O
found	O
between	O
tumor	B-Cancer
prognosis	O
and	O
EGFR	B-Gene_or_gene_product
,	O
ICAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
or	O
Fas	B-Gene_or_gene_product
.	O

However	O
,	O
a	O
statistically	O
significant	O
correlation	O
was	O
found	O
between	O
the	O
surface	B-Cellular_component
expression	O
of	O
CD40	B-Gene_or_gene_product
and	O
the	O
metastatic	O
spread	O
of	O
the	O
tumor	B-Cancer
.	O

In	O
this	O
study	O
,	O
14	O
of	O
18	O
lung	B-Cell
cancer	I-Cell
cell	I-Cell
lines	I-Cell
(	O
78	O
%	O
)	O
expressed	O
CD40	B-Gene_or_gene_product
on	O
their	O
surface	B-Cellular_component
.	O

All	O
of	O
the	O
4	O
tumors	B-Cancer
that	O
were	O
CD40	B-Gene_or_gene_product
-	O
negative	O
,	O
were	O
stage	O
I	O
tumors	B-Cancer
,	O
without	O
any	O
evidence	O
of	O
regional	O
or	O
distant	O
metastasis	O
.	O

Of	O
the	O
14	O
tumors	B-Cancer
that	O
expressed	O
CD40	B-Gene_or_gene_product
,	O
all	O
but	O
1	O
(	O
93	O
%	O
)	O
had	O
either	O
nodal	B-Multi-tissue_structure
or	O
systemic	O
metastasis	O
at	O
the	O
time	O
of	O
diagnosis	O
.	O

Patients	B-Organism
whose	O
tumors	B-Cancer
were	O
CD40	B-Gene_or_gene_product
-	O
negative	O
showed	O
a	O
significantly	O
better	O
N	O
stage	O
,	O
overall	O
stage	O
at	O
presentation	O
and	O
survival	O
than	O
those	O
patients	B-Organism
with	O
CD40	B-Gene_or_gene_product
-	O
positive	O
patients	B-Organism
.	O

No	O
significant	O
differences	O
between	O
the	O
two	O
groups	O
were	O
observed	O
in	O
tumor	B-Cancer
size	O
,	O
gender	O
,	O
age	O
,	O
histology	O
,	O
differentiation	O
or	O
preoperative	O
therapy	O
.	O

CONCLUSIONS	O
:	O
These	O
results	O
suggest	O
that	O
CD40	B-Gene_or_gene_product
expression	O
on	O
lung	B-Cancer
cancer	I-Cancer
may	O
play	O
a	O
role	O
in	O
metastatic	O
spread	O
,	O
and	O
also	O
may	O
serve	O
as	O
a	O
prognostic	O
marker	O
and	O
an	O
indicator	O
of	O
advanced	O
disease	O
.	O

The	O
value	O
of	O
serum	B-Organism_substance
S	B-Gene_or_gene_product
-	I-Gene_or_gene_product
100beta	I-Gene_or_gene_product
and	O
interleukins	B-Gene_or_gene_product
as	O
tumour	B-Cancer
markers	O
in	O
advanced	O
melanoma	B-Cancer
.	O

Recently	O
serum	B-Organism_substance
S	B-Gene_or_gene_product
-	I-Gene_or_gene_product
100beta	I-Gene_or_gene_product
has	O
shown	O
promise	O
as	O
a	O
tumour	B-Cancer
marker	O
in	O
melanoma	B-Cancer
;	O
however	O
,	O
its	O
use	O
as	O
a	O
prognostic	O
marker	O
in	O
the	O
advanced	O
stage	O
needs	O
to	O
be	O
confirmed	O
.	O

Interleukins	B-Gene_or_gene_product
(	O
ILs	B-Gene_or_gene_product
)	O
may	O
mediate	O
regression	O
or	O
progression	O
of	O
cancer	B-Cancer
.	O

In	O
order	O
to	O
study	O
their	O
relation	O
to	O
the	O
metastatic	O
profile	O
and	O
survival	O
,	O
we	O
evaluated	O
the	O
association	O
between	O
pretreatment	O
serum	B-Organism_substance
levels	O
of	O
S	B-Gene_or_gene_product
-	I-Gene_or_gene_product
100beta	I-Gene_or_gene_product
,	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
6	I-Gene_or_gene_product
,	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
10	I-Gene_or_gene_product
and	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
12	I-Gene_or_gene_product
and	O
metastatic	O
site	O
and	O
survival	O
in	O
50	O
patients	B-Organism
with	O
advanced	O
melanoma	B-Cancer
who	O
were	O
to	O
receive	O
chemoimmunotherapy	O
.	O

Patients	B-Organism
with	O
liver	B-Organ
and	O
/	O
or	O
bone	B-Organ
metastases	O
had	O
significantly	O
higher	O
median	O
concentrations	O
of	O
S	B-Gene_or_gene_product
-	I-Gene_or_gene_product
100beta	I-Gene_or_gene_product
,	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
6	I-Gene_or_gene_product
and	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
10	I-Gene_or_gene_product
than	O
those	O
with	O
only	O
skin	B-Organ
,	O
nodal	B-Multi-tissue_structure
and	O
/	O
or	O
lung	B-Organ
involvement	O
.	O

The	O
differences	O
in	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
12	I-Gene_or_gene_product
levels	O
were	O
unremarkable	O
.	O

Using	O
univariate	O
analysis	O
,	O
the	O
S	B-Gene_or_gene_product
-	I-Gene_or_gene_product
100beta	I-Gene_or_gene_product
level	O
and	O
metastatic	O
profile	O
were	O
found	O
to	O
be	O
statistically	O
significant	O
prognostic	O
factors	O
for	O
survival	O
.	O

Using	O
multivariate	O
analysis	O
the	O
S	B-Gene_or_gene_product
-	I-Gene_or_gene_product
100beta	I-Gene_or_gene_product
level	O
was	O
the	O
most	O
powerful	O
prognostic	O
indicator	O
,	O
while	O
the	O
metastatic	O
profile	O
was	O
found	O
to	O
be	O
significant	O
after	O
exclusion	O
of	O
S	B-Gene_or_gene_product
-	I-Gene_or_gene_product
100beta	I-Gene_or_gene_product
.	O

The	O
findings	O
suggest	O
that	O
elevated	O
serum	B-Organism_substance
levels	O
of	O
S	B-Gene_or_gene_product
-	I-Gene_or_gene_product
100beta	I-Gene_or_gene_product
,	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
6	I-Gene_or_gene_product
and	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
10	I-Gene_or_gene_product
reflect	O
concurrent	O
liver	B-Organ
or	O
bone	B-Organ
metastases	O
in	O
melanoma	B-Cancer
.	O

S	B-Gene_or_gene_product
-	I-Gene_or_gene_product
100beta	I-Gene_or_gene_product
is	O
also	O
an	O
independent	O
prognostic	O
marker	O
.	O

Pretreatment	O
IL	B-Gene_or_gene_product
levels	O
were	O
not	O
associated	O
with	O
outcome	O
.	O

TEL	B-Gene_or_gene_product
,	O
a	O
putative	O
tumor	B-Cancer
suppressor	O
,	O
modulates	O
cell	B-Cell
growth	O
and	O
cell	B-Cell
morphology	O
of	O
ras	B-Gene_or_gene_product
-	O
transformed	O
cells	B-Cell
while	O
repressing	O
the	O
transcription	O
of	O
stromelysin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

TEL	B-Gene_or_gene_product
is	O
a	O
member	O
of	O
the	O
ETS	B-Gene_or_gene_product
family	O
of	O
transcription	O
factors	O
that	O
interacts	O
with	O
the	O
mSin3	B-Gene_or_gene_product
and	O
SMRT	B-Gene_or_gene_product
corepressors	O
to	O
regulate	O
transcription	O
.	O

TEL	B-Gene_or_gene_product
is	O
biallelically	O
disrupted	O
in	O
acute	B-Cancer
leukemia	I-Cancer
,	O
and	O
loss	O
of	O
heterozygosity	O
at	O
the	O
TEL	B-Gene_or_gene_product
locus	O
has	O
been	O
observed	O
in	O
various	O
cancers	B-Cancer
.	O

Here	O
we	O
show	O
that	O
expression	O
of	O
TEL	B-Gene_or_gene_product
in	O
Ras	B-Gene_or_gene_product
-	O
transformed	O
NIH	B-Cell
3T3	I-Cell
cells	I-Cell
inhibits	O
cell	B-Cell
growth	O
in	O
soft	O
agar	O
and	O
in	O
normal	O
cultures	O
.	O

Unexpectedly	O
,	O
cells	B-Cell
expressing	O
both	O
Ras	B-Gene_or_gene_product
and	O
TEL	B-Gene_or_gene_product
grew	O
as	O
aggregates	O
.	O

To	O
begin	O
to	O
explain	O
the	O
morphology	O
of	O
Ras	B-Gene_or_gene_product
-	O
plus	O
TEL	B-Gene_or_gene_product
-	O
expressing	O
cells	B-Cell
,	O
we	O
demonstrated	O
that	O
the	O
endogenous	O
matrix	B-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
stromelysin	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
was	O
repressed	O
by	O
TEL	B-Gene_or_gene_product
.	O

TEL	B-Gene_or_gene_product
bound	O
sequences	O
in	O
the	O
stromelysin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
promoter	O
and	O
repressed	O
the	O
promoter	O
in	O
transient	O
-	O
expression	O
assays	O
,	O
suggesting	O
that	O
it	O
is	O
a	O
direct	O
target	O
for	O
TEL	B-Gene_or_gene_product
-	O
mediated	O
regulation	O
.	O

Mutants	O
of	O
TEL	B-Gene_or_gene_product
that	O
removed	O
a	O
binding	O
site	O
for	O
the	O
mSin3A	B-Gene_or_gene_product
corepressor	O
but	O
retained	O
the	O
ETS	O
domain	O
failed	O
to	O
repress	O
stromelysin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

When	O
BB	B-Simple_chemical
-	I-Simple_chemical
94	I-Simple_chemical
,	O
a	O
matrix	B-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
inhibitor	O
,	O
was	O
added	O
to	O
the	O
culture	O
medium	O
of	O
Ras	B-Gene_or_gene_product
-	O
expressing	O
cells	B-Cell
,	O
it	O
caused	O
a	O
cell	B-Cell
aggregation	O
phenotype	O
similar	O
to	O
that	O
caused	O
by	O
TEL	B-Gene_or_gene_product
expression	O
.	O

In	O
addition	O
,	O
TEL	B-Gene_or_gene_product
inhibited	O
the	O
invasiveness	O
of	O
Ras	B-Gene_or_gene_product
-	O
transformed	O
cells	B-Cell
in	O
vitro	O
and	O
in	O
vivo	O
.	O

Our	O
results	O
suggest	O
that	O
TEL	B-Gene_or_gene_product
acts	O
as	O
a	O
tumor	B-Cancer
suppressor	O
,	O
in	O
part	O
,	O
by	O
transcriptional	O
repression	O
of	O
stromelysin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

Caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
is	O
essential	O
for	O
procaspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
processing	O
and	O
cisplatin	B-Simple_chemical
-	O
induced	O
apoptosis	O
of	O
MCF	B-Cell
-	I-Cell
7	I-Cell
breast	I-Cell
cancer	I-Cell
cells	I-Cell
.	O

In	O
this	O
study	O
,	O
we	O
sought	O
to	O
investigate	O
in	O
more	O
detail	O
the	O
role	O
of	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
in	O
apoptotic	O
processes	O
in	O
cultured	O
cells	B-Cell
and	O
in	O
cell	B-Cell
-	O
free	O
extracts	B-Organism_substance
of	O
breast	B-Cell
cancer	I-Cell
cells	I-Cell
.	O

We	O
present	O
evidence	O
that	O
apoptosis	O
of	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
-	O
deficient	O
MCF	O
-	O
7	O
breast	O
cancer	O
cells	O
is	O
defective	O
in	O
response	O
to	O
cisplatin	B-Simple_chemical
treatment	O
,	O
as	O
determined	O
by	O
chromatin	B-Cellular_component
condensation	O
,	O
nuclear	B-Cellular_component
fragmentation	O
,	O
DNA	B-Cellular_component
fragmentation	O
,	O
and	O
release	O
of	O
cytochrome	B-Gene_or_gene_product
c	I-Gene_or_gene_product
from	O
the	O
mitochondria	B-Cellular_component
.	O

Reconstitution	O
of	O
MCF	B-Cell
-	I-Cell
7	I-Cell
cells	I-Cell
by	O
stable	O
transfection	O
of	O
CASP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
cDNA	O
restores	O
all	O
these	O
defects	O
and	O
results	O
in	O
an	O
extensive	O
apoptosis	O
after	O
cisplatin	B-Simple_chemical
treatment	O
.	O

We	O
further	O
show	O
that	O
in	O
extracts	B-Organism_substance
from	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
-	O
deficient	O
MCF	O
-	O
7	O
cells	O
,	O
procaspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
processing	O
is	O
strongly	O
impaired	O
after	O
stimulation	O
with	O
either	O
cytochrome	B-Gene_or_gene_product
c	I-Gene_or_gene_product
or	O
recombinant	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
.	O

Reconstitution	O
of	O
MCF	B-Organism_substance
-	I-Organism_substance
7	I-Organism_substance
cell	I-Organism_substance
extracts	I-Organism_substance
with	O
procaspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
corrects	O
this	O
defect	O
,	O
resulting	O
in	O
an	O
efficient	O
and	O
complete	O
processing	O
of	O
procaspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
.	O

Together	O
,	O
our	O
data	O
define	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
as	O
an	O
important	O
integrator	O
of	O
the	O
apoptotic	O
process	O
in	O
MCF	B-Cell
-	I-Cell
7	I-Cell
breast	I-Cell
cancer	I-Cell
cells	I-Cell
and	O
reveal	O
an	O
essential	O
function	O
of	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
for	O
procaspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
processing	O
.	O

Vasodilator	B-Gene_or_gene_product
-	I-Gene_or_gene_product
stimulated	I-Gene_or_gene_product
phosphoprotein	I-Gene_or_gene_product
is	O
involved	O
in	O
stress	B-Cellular_component
-	I-Cellular_component
fiber	I-Cellular_component
and	O
membrane	B-Cellular_component
ruffle	I-Cellular_component
formation	O
in	O
endothelial	B-Cell
cells	I-Cell
.	O

Vasodilator	B-Gene_or_gene_product
-	I-Gene_or_gene_product
stimulated	I-Gene_or_gene_product
phosphoprotein	I-Gene_or_gene_product
(	O
VASP	B-Gene_or_gene_product
)	O
is	O
highly	O
expressed	O
in	O
vascular	B-Cell
endothelial	I-Cell
cells	I-Cell
,	O
where	O
it	O
has	O
been	O
implicated	O
in	O
cellular	B-Cell
reorganization	O
during	O
angiogenesis	O
,	O
as	O
well	O
as	O
in	O
endothelial	B-Cell
retraction	O
and	O
changes	O
in	O
vessel	B-Multi-tissue_structure
permeability	O
.	O

However	O
,	O
the	O
cellular	B-Cell
functions	O
of	O
VASP	B-Gene_or_gene_product
are	O
not	O
known	O
.	O

In	O
this	O
study	O
,	O
we	O
have	O
expressed	O
wild	O
-	O
type	O
and	O
mutant	O
forms	O
of	O
VASP	B-Gene_or_gene_product
in	O
endothelial	B-Cell
cells	I-Cell
to	O
determine	O
in	O
what	O
aspects	O
of	O
cytoskeletal	B-Cellular_component
behavior	O
this	O
protein	O
participates	O
.	O

Expression	O
of	O
wild	O
-	O
type	O
VASP	B-Gene_or_gene_product
induces	O
marked	O
membrane	B-Cellular_component
ruffling	O
and	O
formation	O
of	O
prominent	O
stress	B-Cellular_component
fibers	I-Cellular_component
in	O
bovine	B-Organism
aortic	B-Cell
endothelial	I-Cell
cells	I-Cell
.	O

Deletion	O
of	O
the	O
proline	O
-	O
rich	O
domain	O
of	O
VASP	B-Gene_or_gene_product
abolishes	O
its	O
ability	O
to	O
bind	O
profilin	B-Gene_or_gene_product
but	O
does	O
not	O
affect	O
ruffling	O
or	O
stress	B-Cellular_component
fiber	I-Cellular_component
formation	O
.	O

Further	O
deletions	O
reveal	O
a	O
sequence	O
within	O
the	O
carboxy	O
-	O
terminal	O
domain	O
that	O
is	O
responsible	O
for	O
in	O
vivo	O
bundle	O
formation	O
.	O

Ruffling	O
occurs	O
only	O
on	O
the	O
expression	O
of	O
forms	O
of	O
VASP	B-Gene_or_gene_product
that	O
possess	O
bundling	O
activity	O
and	O
the	O
capacity	O
to	O
bind	O
zyxin	B-Gene_or_gene_product
/	O
vinculin	B-Gene_or_gene_product
-	O
derived	O
peptide	O
.	O

The	O
ability	O
of	O
distinct	O
subdomains	O
within	O
VASP	B-Gene_or_gene_product
to	O
bind	O
adhesion	O
proteins	O
and	O
induce	O
F	B-Gene_or_gene_product
-	I-Gene_or_gene_product
actin	I-Gene_or_gene_product
bundling	O
in	O
vivo	O
suggests	O
that	O
this	O
protein	O
could	O
function	O
in	O
the	O
aggregation	O
and	O
tethering	O
of	O
actin	B-Gene_or_gene_product
filaments	O
during	O
the	O
formation	O
of	O
endothelial	B-Cellular_component
cell	I-Cellular_component
-	I-Cellular_component
substrate	I-Cellular_component
and	O
cell	B-Cellular_component
-	I-Cellular_component
cell	I-Cellular_component
contacts	I-Cellular_component
.	O

These	O
data	O
provide	O
a	O
mechanism	O
whereby	O
VASP	B-Gene_or_gene_product
can	O
influence	O
endothelial	B-Cell
migration	O
and	O
organization	O
during	O
capillary	B-Tissue
formation	O
and	O
modulate	O
vascular	B-Multi-tissue_structure
permeability	O
via	O
effects	O
on	O
endothelial	B-Cell
cell	I-Cell
contractility	O
.	O

Urokinase	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
:	O
a	O
molecular	O
organizer	O
in	O
cellular	B-Cell
communication	O
.	O

In	O
a	O
variety	O
of	O
cell	B-Cell
types	O
,	O
the	O
glycolipid	B-Gene_or_gene_product
-	I-Gene_or_gene_product
anchored	I-Gene_or_gene_product
urokinase	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
(	O
uPAR	B-Gene_or_gene_product
)	O
is	O
colocalized	O
pericellularly	O
with	O
components	O
of	O
the	O
plasminogen	B-Gene_or_gene_product
activation	O
system	O
and	O
endocytosis	O
receptors	O
.	O

uPAR	B-Gene_or_gene_product
is	O
also	O
coexpressed	O
with	O
caveolin	B-Gene_or_gene_product
and	O
members	O
of	O
the	O
integrin	B-Gene_or_gene_product
adhesion	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
superfamily	O
.	O

The	O
formation	O
of	O
functional	O
units	O
with	O
these	O
various	O
proteins	O
allows	O
the	O
uPAR	B-Gene_or_gene_product
to	O
mediate	O
the	O
focused	O
proteolysis	O
required	O
for	O
cell	B-Cell
migration	O
and	O
invasion	O
and	O
to	O
contribute	O
both	O
directly	O
and	O
indirectly	O
to	O
cell	B-Cell
adhesive	O
processes	O
in	O
a	O
non	O
-	O
proteolytic	O
fashion	O
.	O

This	O
dual	O
activity	O
,	O
together	O
with	O
the	O
initiation	O
of	O
signal	O
transduction	O
pathways	O
by	O
uPAR	B-Gene_or_gene_product
,	O
is	O
believed	O
to	O
influence	O
cellular	B-Cell
behaviour	O
in	O
angiogenesis	O
,	O
inflammation	O
,	O
wound	B-Pathological_formation
repair	O
and	O
tumor	B-Cancer
progression	O
/	O
metastasis	O
and	O
open	O
up	O
the	O
way	O
for	O
uPAR	B-Gene_or_gene_product
-	O
based	O
therapeutic	O
approaches	O
.	O

p75	B-Gene_or_gene_product
mediated	O
apoptosis	O
in	O
neuroblastoma	B-Cell
cells	I-Cell
is	O
inhibited	O
by	O
expression	O
of	O
TrkA	B-Gene_or_gene_product
.	O

BACKGROUND	O
:	O
Neurotrophins	B-Gene_or_gene_product
mediate	O
their	O
effects	O
by	O
binding	O
to	O
members	O
of	O
the	O
Trk	B-Gene_or_gene_product
family	O
of	O
receptor	O
tyrosine	O
kinases	O
and	O
to	O
the	O
low	O
-	O
affinity	O
nerve	O
growth	O
factor	O
receptor	O
p75	B-Gene_or_gene_product
.	O

Nerve	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
NGF	B-Gene_or_gene_product
)	O
has	O
been	O
demonstrated	O
to	O
support	O
survival	O
and	O
differentiation	O
of	O
neuroblastoma	B-Cell
(	I-Cell
NB	I-Cell
)	I-Cell
cells	I-Cell
by	O
activation	O
of	O
the	O
TrkA	B-Gene_or_gene_product
receptor	O
.	O

The	O
p75	B-Gene_or_gene_product
receptor	O
belongs	O
to	O
the	O
tumor	B-Gene_or_gene_product
necrosis	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
TNF	B-Gene_or_gene_product
)	O
family	O
of	O
death	O
receptors	O
and	O
has	O
been	O
suggested	O
as	O
a	O
receptor	O
that	O
mediates	O
apoptosis	O
in	O
neuronal	B-Cell
and	O
NB	B-Cell
cells	I-Cell
.	O

PROCEDURE	O
:	O
To	O
investigate	O
the	O
effect	O
of	O
p75	B-Gene_or_gene_product
expression	O
in	O
NB	B-Cancer
,	O
we	O
transfected	O
the	O
p75	B-Gene_or_gene_product
cDNA	O
into	O
SH	B-Cell
-	I-Cell
SY5Y	I-Cell
cells	I-Cell
,	O
an	O
NB	B-Cell
cell	I-Cell
line	I-Cell
lacking	O
expression	O
of	O
both	O
p75	B-Gene_or_gene_product
and	O
TrkA	B-Gene_or_gene_product
.	O

RESULTS	O
:	O
Cell	B-Cell
clones	I-Cell
expressing	O
elevated	O
levels	O
of	O
p75	B-Gene_or_gene_product
showed	O
a	O
high	O
degree	O
of	O
apoptosis	O
even	O
in	O
10	O
%	O
serum	B-Organism_substance
-	O
supplemented	O
medium	O
.	O

Apoptotic	O
signaling	O
by	O
p75	B-Gene_or_gene_product
was	O
ligand	O
-	O
independent	O
and	O
only	O
partly	O
caspase	B-Gene_or_gene_product
-	O
dependent	O
.	O

The	O
level	O
of	O
apoptosis	O
correlated	O
directly	O
with	O
the	O
expression	O
level	O
of	O
the	O
receptor	O
,	O
indicating	O
that	O
p75	B-Gene_or_gene_product
may	O
activate	O
the	O
cell	B-Cell
death	O
program	O
directly	O
.	O

However	O
,	O
additional	O
transfection	O
of	O
TrkA	B-Gene_or_gene_product
into	O
SY5Y	B-Gene_or_gene_product
-	O
p75	B-Gene_or_gene_product
cells	O
resulted	O
in	O
a	O
significantly	O
reduced	O
rate	O
of	O
apoptosis	O
even	O
in	O
the	O
absence	O
of	O
NGF	B-Gene_or_gene_product
.	O

CONCLUSIONS	O
:	O
Thus	O
,	O
expression	O
of	O
the	O
TrkA	B-Gene_or_gene_product
receptor	O
itself	O
inhibits	O
p75	B-Gene_or_gene_product
mediated	O
apoptosis	O
in	O
NB	B-Cell
cells	I-Cell
.	O

RNA	O
damage	O
and	O
inhibition	O
of	O
neoplastic	B-Cell
endothelial	I-Cell
cell	I-Cell
growth	O
:	O
effects	O
of	O
human	B-Organism
and	O
amphibian	O
ribonucleases	B-Gene_or_gene_product
.	O

Angiogenesis	O
defines	O
the	O
many	O
steps	O
involved	O
in	O
the	O
growth	O
and	O
migration	O
of	O
endothelial	B-Cell
cell	I-Cell
-	O
derived	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
.	O

This	O
process	O
is	O
necessary	O
for	O
the	O
growth	O
and	O
metastasis	O
of	O
tumors	B-Cancer
,	O
and	O
considerable	O
effort	O
is	O
being	O
expended	O
to	O
find	O
inhibitors	O
of	O
tumor	B-Cancer
angiogenesis	O
.	O

This	O
usually	O
involves	O
screening	O
of	O
potential	O
anti	O
-	O
angiogenic	O
compounds	O
on	O
endothelial	B-Cell
cells	I-Cell
.	O

To	O
this	O
end	O
,	O
two	O
candidate	O
anti	O
-	O
angiogenic	O
RNA	O
-	O
damaging	O
agents	O
,	O
onconase	B-Simple_chemical
and	O
(	B-Simple_chemical
-	I-Simple_chemical
4	I-Simple_chemical
)	I-Simple_chemical
rhEDN	I-Simple_chemical
,	O
were	O
screened	O
for	O
their	O
effects	O
on	O
endothelial	B-Cell
cell	I-Cell
proliferation	O
using	O
three	O
distinct	O
types	O
of	O
endothelial	B-Cell
cells	I-Cell
in	O
culture	O
:	O
HPV	B-Organism
-	I-Organism
16	I-Organism
E6	I-Organism
/	O
E7	B-Organism
-	O
immortalized	O
human	B-Cell
umbilical	I-Cell
vein	I-Cell
endothelial	I-Cell
cells	I-Cell
(	O
HUVECs	B-Cell
)	O
,	O
a	O
Kras	B-Gene_or_gene_product
-	O
transformed	O
HPV	B-Organism
-	I-Organism
16	I-Organism
E6	I-Organism
/	O
E7	B-Organism
HUVEC	B-Cell
(	O
Rhim	O
et	O
al	O
.	O
,	O
Carcinogenesis	O
4	O
,	O
673	O
-	O
681	O
,	O
1998	O
)	O
,	O
and	O
primary	O
HUVECs	B-Cell
.	O

Onconase	B-Simple_chemical
similarly	O
inhibited	O
proliferation	O
in	O
all	O
three	O
cell	B-Cell
lines	I-Cell
(	O
IC	O
(	O
50	O
)	O
=	O
0	O
.	O
3	O
-	O
1	O
.	O
0	O
microM	O
)	O
while	O
(	B-Simple_chemical
-	I-Simple_chemical
4	I-Simple_chemical
)	I-Simple_chemical
rhEDN	I-Simple_chemical
was	O
more	O
effective	O
on	O
immortalized	O
HUVEC	B-Cell
cell	I-Cell
lines	I-Cell
(	O
IC	O
(	O
50	O
)	O
=	O
0	O
.	O
02	O
-	O
0	O
.	O
06	O
microM	O
)	O
than	O
on	O
primary	O
HUVECs	B-Cell
(	O
IC	O
(	O
50	O
)	O
>	O
0	O
.	O
1	O
microM	O
)	O
.	O

Differential	O
sensitivity	O
to	O
these	O
agents	O
implies	O
that	O
more	O
than	O
one	O
endothelial	B-Cell
cell	I-Cell
type	I-Cell
must	O
be	O
used	O
in	O
proliferation	O
assays	O
to	O
screen	O
for	O
novel	O
anti	O
-	O
angiogenic	O
compounds	O
.	O

TRADD	B-Gene_or_gene_product
domain	O
of	O
Epstein	B-Organism
-	I-Organism
Barr	I-Organism
virus	I-Organism
transforming	O
protein	O
LMP1	B-Gene_or_gene_product
is	O
essential	O
for	O
inducing	O
immortalization	O
and	O
suppressing	O
senescence	O
of	O
primary	O
rodent	B-Organism
fibroblasts	B-Cell
.	O

Mutation	O
analysis	O
of	O
latent	B-Gene_or_gene_product
membrane	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
LMP1	B-Gene_or_gene_product
)	O
in	O
Epstein	B-Organism
-	I-Organism
Barr	I-Organism
virus	I-Organism
(	O
EBV	B-Organism
)	O
-	O
induced	O
B	B-Cell
-	I-Cell
cell	I-Cell
immortalization	O
revealed	O
two	O
transformation	O
effector	O
sites	O
,	O
TES1	O
and	O
TES2	O
.	O

TES2	O
mediates	O
the	O
interaction	O
with	O
tumor	B-Gene_or_gene_product
necrosis	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
associated	I-Gene_or_gene_product
death	I-Gene_or_gene_product
domain	I-Gene_or_gene_product
protein	O
(	O
TRADD	B-Gene_or_gene_product
)	O
and	O
plays	O
a	O
key	O
role	O
in	O
transactivating	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappa	I-Gene_or_gene_product
B	I-Gene_or_gene_product
and	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

Recombinant	O
EBV	B-Organism
containing	O
LMP1	B-Gene_or_gene_product
with	O
TES2	O
deleted	O
induces	O
a	O
limited	O
proliferation	O
of	O
B	B-Cell
cells	I-Cell
.	O

The	O
present	O
study	O
shows	O
that	O
a	O
mutant	O
with	O
an	O
LMP1	B-Gene_or_gene_product
site	O
-	O
specific	O
mutation	O
at	O
TES2	O
,	O
LMP1	B-Gene_or_gene_product
(	O
TRADD	B-Gene_or_gene_product
)	O
,	O
initially	O
stimulates	O
cell	B-Cell
growth	O
and	O
significantly	O
extends	O
the	O
life	O
span	O
of	O
MEF	B-Cell
.	O

However	O
,	O
it	O
is	O
not	O
sufficient	O
for	O
the	O
immortalization	O
of	O
MEF	B-Cell
,	O
and	O
MEF	O
-	O
LMP1	B-Gene_or_gene_product
(	O
TRADD	B-Gene_or_gene_product
)	O
cells	O
eventually	O
enter	O
growth	O
arrest	O
.	O

Further	O
analysis	O
reveals	O
that	O
although	O
LMP1	B-Gene_or_gene_product
(	O
TRADD	B-Gene_or_gene_product
)	O
promotes	O
cell	B-Cell
growth	O
,	O
it	O
does	O
not	O
prevent	O
the	O
eventual	O
onset	O
of	O
senescence	O
and	O
the	O
expression	O
of	O
tumor	B-Cancer
suppressor	O
p16	B-Gene_or_gene_product
(	O
Ink4a	B-Gene_or_gene_product
)	O
.	O

IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
12	I-Gene_or_gene_product
inhibition	O
of	O
endothelial	B-Cell
cell	I-Cell
functions	O
and	O
angiogenesis	O
depends	O
on	O
lymphocyte	B-Cell
-	O
endothelial	B-Cell
cell	I-Cell
cross	O
-	O
talk	O
.	O

In	O
vivo	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
12	I-Gene_or_gene_product
-	O
dependent	O
tumor	B-Cancer
inhibition	O
rests	O
on	O
the	O
ability	O
of	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
12	I-Gene_or_gene_product
to	O
activate	O
a	O
CD8	B-Gene_or_gene_product
-	O
mediated	O
cytotoxicity	O
,	O
inhibit	O
angiogenesis	O
,	O
and	O
cause	O
vascular	B-Multi-tissue_structure
injury	O
.	O

Although	O
in	O
vivo	O
studies	O
have	O
shown	O
that	O
such	O
inhibition	O
stems	O
from	O
complex	O
interactions	O
of	O
immune	B-Cell
cells	I-Cell
and	O
the	O
production	O
of	O
IFN	B-Gene_or_gene_product
-	I-Gene_or_gene_product
gamma	I-Gene_or_gene_product
and	O
other	O
downstream	O
angiostatic	O
chemokines	O
,	O
the	O
mechanisms	O
involved	O
are	O
still	O
poorly	O
defined	O
.	O

Here	O
we	O
show	O
that	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
12	I-Gene_or_gene_product
activates	O
an	O
anti	O
-	O
angiogenic	O
program	O
in	O
Con	B-Simple_chemical
A	I-Simple_chemical
-	O
activated	O
mouse	B-Organism
spleen	B-Cell
cells	I-Cell
(	O
activated	O
spc	B-Cell
)	O
or	O
human	B-Organism
PBMC	B-Cell
(	O
activated	O
PBMC	B-Cell
)	O
.	O

The	O
soluble	O
factors	O
they	O
release	O
in	O
its	O
presence	O
arrest	O
the	O
cycle	O
of	O
endothelial	B-Cell
cells	I-Cell
(	O
EC	B-Cell
)	O
,	O
inhibit	O
in	O
vitro	O
angiogenesis	O
,	O
negatively	O
modulate	O
the	O
production	O
of	O
matrix	B-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
,	O
and	O
the	O
ability	O
of	O
EC	B-Cell
to	O
adhere	O
to	O
vitronectin	B-Gene_or_gene_product
and	O
up	O
-	O
regulate	O
ICAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
VCAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
expression	O
.	O

These	O
effects	O
do	O
not	O
require	O
direct	O
cell	B-Cell
-	O
cell	B-Cell
contact	O
,	O
yet	O
result	O
from	O
continuous	O
interaction	O
between	O
activated	O
lymphoid	B-Cell
cells	I-Cell
and	O
EC	B-Cell
.	O

We	O
used	O
neutralizing	O
Abs	O
to	O
show	O
that	O
the	O
IFN	B-Gene_or_gene_product
-	I-Gene_or_gene_product
inducible	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
-	I-Gene_or_gene_product
10	I-Gene_or_gene_product
and	O
monokine	O
-	O
induced	O
by	O
IFN	O
-	O
gamma	O
chemokines	O
are	O
pivotal	O
in	O
inducing	O
these	O
effects	O
.	O

Experiments	O
with	O
nu	B-Organism
/	I-Organism
nu	I-Organism
mice	I-Organism
,	O
nonobese	B-Organism
diabetic	I-Organism
-	I-Organism
SCID	I-Organism
mice	I-Organism
,	O
or	O
activated	O
spc	B-Cell
enriched	O
in	O
specific	O
cell	B-Cell
subpopulations	O
demonstrated	O
that	O
CD4	B-Gene_or_gene_product
(	O
+	O
)	O
,	O
CD8	B-Gene_or_gene_product
(	O
+	O
)	O
,	O
and	O
NK	B-Cell
cells	I-Cell
are	O
all	O
needed	O
to	O
mediate	O
the	O
full	O
anti	O
-	O
angiogenetic	O
effect	O
of	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
12	I-Gene_or_gene_product
.	O

[	O
Study	O
of	O
fungus	B-Simple_chemical
polysaccharides	I-Simple_chemical
compounds	I-Simple_chemical
(	O
FPC	B-Simple_chemical
)	O
in	O
inducing	O
the	O
apoptosis	O
of	O
liver	B-Cell
cancer	I-Cell
cell	I-Cell
Bel	I-Cell
-	I-Cell
7402	I-Cell
]	I-Cell
.	O

To	O
observe	O
the	O
influence	O
of	O
fungus	B-Simple_chemical
polysaccharides	I-Simple_chemical
compounds	I-Simple_chemical
(	O
FPC	B-Simple_chemical
)	O
in	O
inducing	O
human	B-Organism
liver	B-Cell
cancer	I-Cell
cell	I-Cell
Bel	I-Cell
-	I-Cell
7402	I-Cell
apoptosis	O
in	O
cell	B-Cell
cultivating	O
in	O
vitro	O
,	O
the	O
authors	O
analyzed	O
tumor	B-Cancer
inhibitive	O
gene	O
P53	B-Gene_or_gene_product
expression	O
in	O
Bel	B-Cell
-	I-Cell
7402	I-Cell
apoptosis	O
by	O
applying	O
double	O
immuno	O
-	O
marker	O
.	O

The	O
result	O
showed	O
that	O
the	O
multilevel	O
of	O
FPC	B-Simple_chemical
could	O
all	O
apparently	O
induce	O
Bel	B-Cell
-	I-Cell
7402	I-Cell
apoptosis	O
.	O

With	O
the	O
enhancement	O
of	O
FPC	B-Simple_chemical
concentration	O
,	O
the	O
authors	O
observed	O
chromatin	B-Cellular_component
condensation	O
in	O
some	O
phases	O
companying	O
with	O
the	O
characteristic	O
apoptosis	O
.	O

In	O
the	O
meantime	O
,	O
it	O
could	O
also	O
greatly	O
reduce	O
the	O
G1	O
and	O
S	O
,	O
with	O
obviously	O
dose	O
-	O
response	O
relationship	O
.	O

The	O
percentage	O
of	O
cell	B-Cell
apoptosis	O
increased	O
with	O
the	O
enhancing	O
of	O
concentration	O
.	O

In	O
the	O
high	O
-	O
level	O
group	O
the	O
authors	O
found	O
typical	O
DNA	B-Cellular_component
ladder	O
eletrophoresis	O
stripe	O
.	O

The	O
result	O
showed	O
that	O
the	O
mechanism	O
of	O
the	O
FPC	B-Simple_chemical
antineoplastic	O
effect	O
had	O
an	O
intimate	O
relation	O
with	O
its	O
induction	O
to	O
apoptosis	O
and	O
that	O
the	O
result	O
of	O
FPC	B-Simple_chemical
inducing	O
tumor	B-Cell
cell	I-Cell
apoptosis	O
had	O
the	O
character	O
of	O
P53	B-Gene_or_gene_product
independence	O
.	O

Effects	O
of	O
morphological	O
patterning	O
on	O
endothelial	B-Cell
cell	I-Cell
migration	O
.	O

The	O
migration	O
of	O
vascular	B-Cell
endothelial	I-Cell
cells	I-Cell
(	O
ECs	B-Cell
)	O
plays	O
an	O
important	O
role	O
in	O
vascular	B-Multi-tissue_structure
remodeling	O
.	O

Here	O
we	O
studied	O
the	O
effects	O
of	O
cell	B-Cell
morphology	O
on	O
the	O
migration	O
of	O
bovine	B-Organism
aortic	B-Cell
ECs	I-Cell
by	O
culturing	O
cells	B-Cell
on	O
micropatterned	O
strips	O
of	O
collagen	B-Gene_or_gene_product
matrix	B-Cellular_component
(	O
60	O
-	O
,	O
30	O
-	O
,	O
and	O
15	O
-	O
microm	O
wide	O
)	O
.	O

The	O
spreading	O
areas	O
of	O
the	O
cells	B-Cell
on	O
15	O
-	O
and	O
30	O
-	O
microm	O
wide	O
strips	O
were	O
30	O
%	O
lower	O
than	O
those	O
on	O
60	O
-	O
microm	O
wide	O
strips	O
and	O
unpatterned	O
collagen	B-Gene_or_gene_product
.	O

The	O
cells	B-Cell
on	O
15	O
-	O
microm	O
wide	O
strips	O
completely	O
aligned	O
in	O
the	O
direction	O
of	O
the	O
strip	O
,	O
and	O
had	O
significantly	O
lower	O
shape	O
index	O
than	O
those	O
in	O
all	O
other	O
groups	O
.	O

On	O
strips	O
of	O
all	O
widths	O
,	O
ECs	B-Cell
tended	O
to	O
migrate	O
in	O
the	O
direction	O
of	O
strips	O
.	O

ECs	B-Cell
on	O
15	O
-	O
microm	O
wide	O
strips	O
had	O
highest	O
speed	O
,	O
particularly	O
in	O
the	O
direction	O
of	O
the	O
strip	O
.	O

Vinculin	B-Gene_or_gene_product
staining	O
showed	O
that	O
the	O
leading	O
edge	O
of	O
ECs	B-Cell
on	O
15	O
-	O
microm	O
wide	O
strips	O
had	O
focal	O
adhesions	O
that	O
were	O
oriented	O
with	O
their	O
lamellipodial	O
protrusion	O
and	O
the	O
direction	O
of	O
cell	B-Cell
migration	O
;	O
this	O
arrangement	O
of	O
the	O
focal	O
adhesions	O
may	O
promote	O
EC	B-Cell
migration	O
.	O

The	O
present	O
study	O
provides	O
direct	O
evidence	O
on	O
the	O
role	O
of	O
cell	B-Cell
morphology	O
in	O
EC	B-Cell
migration	O
,	O
and	O
will	O
help	O
us	O
to	O
understand	O
the	O
mechanisms	O
of	O
EC	B-Cell
migration	O
during	O
angiogenesis	O
and	O
wound	B-Pathological_formation
healing	O
.	O

Thrombospondin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
plays	O
a	O
protective	O
role	O
in	O
multistep	O
carcinogenesis	O
:	O
a	O
novel	O
host	O
anti	O
-	O
tumor	B-Cancer
defense	O
mechanism	O
.	O

The	O
angiogenic	O
switch	O
during	O
tumorigenesis	O
is	O
thought	O
to	O
be	O
induced	O
by	O
a	O
change	O
in	O
the	O
balance	O
of	O
pro	O
-	O
angiogenic	O
and	O
anti	O
-	O
angiogenic	O
factors	O
.	O

To	O
elucidate	O
the	O
biological	O
role	O
of	O
the	O
endogenous	O
angiogenesis	O
inhibitor	O
thrombospondin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
(	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
)	O
during	O
multistep	O
carcinogenesis	O
,	O
we	O
subjected	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
-	O
deficient	O
and	O
wild	O
-	O
type	O
mice	B-Organism
to	O
a	O
chemical	O
skin	B-Organ
carcinogenesis	O
regimen	O
.	O

Surprisingly	O
,	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
expression	O
was	O
strongly	O
upregulated	O
in	O
the	O
mesenchymal	B-Tissue
stroma	I-Tissue
of	O
wild	O
-	O
type	O
mice	B-Organism
throughout	O
the	O
consecutive	O
stages	O
of	O
tumorigenesis	O
whereas	O
the	O
angiogenesis	O
factor	O
,	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
,	O
was	O
induced	O
predominantly	O
in	O
tumor	B-Cell
cells	I-Cell
.	O

TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
deficiency	O
dramatically	O
enhanced	O
susceptibility	O
to	O
skin	B-Organ
carcinogenesis	O
and	O
resulted	O
in	O
accelerated	O
and	O
increased	O
tumor	B-Cancer
formation	O
.	O

The	O
angiogenic	O
switch	O
occurred	O
in	O
early	O
stages	O
of	O
pre	B-Pathological_formation
-	I-Pathological_formation
malignant	I-Pathological_formation
tumor	I-Pathological_formation
formation	O
,	O
and	O
tumor	B-Cancer
angiogenesis	O
was	O
significantly	O
enhanced	O
in	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
-	O
deficient	O
mice	B-Organism
.	O

While	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
deficiency	O
did	O
not	O
affect	O
tumor	B-Cancer
differentiation	O
or	O
proliferation	O
,	O
tumor	B-Cell
cell	I-Cell
apoptosis	O
was	O
significantly	O
reduced	O
.	O

These	O
results	O
reveal	O
upregulation	O
of	O
an	O
endogenous	O
angiogenesis	O
inhibitor	O
during	O
multi	O
step	O
tumorigenesis	O
and	O
identify	O
enhanced	O
stromal	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
expression	O
as	O
a	O
novel	O
host	O
anti	O
-	O
tumor	B-Cancer
defense	O
mechanism	O
.	O

Relationship	O
of	O
p53	B-Gene_or_gene_product
and	O
Helicobacter	B-Organism
pylori	I-Organism
to	O
clinicopathological	O
features	O
of	O
human	B-Organism
remnant	B-Cancer
stomach	I-Cancer
cancer	I-Cancer
after	O
gastric	B-Organ
surgery	O
for	O
primary	O
gastric	B-Cancer
cancer	I-Cancer
.	O

The	O
purpose	O
of	O
this	O
study	O
was	O
to	O
evaluate	O
the	O
biological	O
features	O
of	O
gastric	B-Cancer
cancer	I-Cancer
of	O
the	O
remnant	B-Organ
stomach	I-Organ
(	O
RSC	B-Cancer
)	O
.	O

Twenty	O
-	O
one	O
patients	B-Organism
underwent	O
resection	O
of	O
the	O
remnant	B-Organ
stomach	I-Organ
for	O
RSC	B-Cancer
and	O
were	O
divided	O
into	O
two	O
groups	O
:	O
the	O
RSCB	B-Cancer
group	O
consisted	O
of	O
11	O
patients	B-Organism
who	O
underwent	O
distal	O
gastrectomy	O
for	O
benign	O
disease	O
and	O
the	O
RSCM	B-Cancer
group	O
consisted	O
of	O
10	O
patients	B-Organism
who	O
underwent	O
gastrectomy	O
for	O
primary	O
gastric	B-Cancer
cancer	I-Cancer
.	O

The	O
interval	O
between	O
primary	O
surgery	O
and	O
the	O
appearance	O
of	O
gastric	B-Cancer
cancer	I-Cancer
in	O
the	O
remnant	B-Organ
stomach	I-Organ
was	O
significantly	O
shorter	O
in	O
the	O
RSCM	B-Cancer
group	O
than	O
in	O
the	O
RSCB	B-Cancer
group	O
.	O

Invasion	O
of	O
adjacent	O
organs	B-Organ
was	O
more	O
frequent	O
in	O
the	O
RSCM	B-Cancer
group	O
than	O
in	O
the	O
RSCB	B-Cancer
group	O
and	O
the	O
Ki	B-Gene_or_gene_product
-	I-Gene_or_gene_product
67	I-Gene_or_gene_product
labeling	O
index	O
of	O
the	O
tumors	B-Cancer
was	O
significantly	O
higher	O
in	O
the	O
former	O
group	O
.	O

Furthermore	O
,	O
p53	B-Gene_or_gene_product
overexpression	O
by	O
tumors	B-Cancer
was	O
almost	O
twice	O
as	O
common	O
in	O
the	O
RSCM	B-Cancer
group	O
as	O
in	O
the	O
RSCB	B-Cancer
group	O
.	O

Although	O
there	O
was	O
no	O
significant	O
difference	O
of	O
the	O
H	B-Organism
.	I-Organism
pylori	I-Organism
positivity	O
between	O
the	O
two	O
groups	O
,	O
the	O
rate	O
for	O
both	O
groups	O
was	O
higher	O
than	O
reported	O
in	O
previous	O
studies	O
.	O

Mutation	O
of	O
p53	B-Gene_or_gene_product
may	O
play	O
an	O
important	O
role	O
in	O
the	O
high	O
proliferative	O
activity	O
of	O
tumors	B-Cancer
in	O
the	O
RSCM	B-Cancer
group	O
and	O
H	B-Organism
.	I-Organism
pylori	I-Organism
infection	O
may	O
be	O
closely	O
related	O
to	O
carcinogenesis	O
in	O
patients	B-Organism
with	O
RSC	B-Cancer
.	O

Current	O
prospects	O
for	O
controlling	O
cancer	B-Cancer
growth	O
with	O
non	O
-	O
cytotoxic	O
agents	O
-	O
-	O
nutrients	O
,	O
phytochemicals	O
,	O
herbal	O
extracts	O
,	O
and	O
available	O
drugs	O
.	O

In	O
animal	O
or	O
cell	B-Cell
culture	O
studies	O
,	O
the	O
growth	O
and	O
spread	O
of	O
cancer	B-Cancer
can	O
be	O
slowed	O
by	O
many	O
nutrients	O
,	O
food	O
factors	O
,	O
herbal	O
extracts	O
,	O
and	O
well	O
-	O
tolerated	O
,	O
available	O
drugs	O
that	O
are	O
still	O
rarely	O
used	O
in	O
the	O
clinical	O
management	O
of	O
cancer	B-Cancer
,	O
in	O
part	O
because	O
they	O
seem	O
unlikely	O
to	O
constitute	O
definitive	O
therapies	O
in	O
themselves	O
.	O

However	O
,	O
it	O
is	O
reasonable	O
to	O
expect	O
that	O
mechanistically	O
complementary	O
combinations	O
of	O
these	O
measures	O
could	O
have	O
a	O
worthwhile	O
impact	O
on	O
survival	O
times	O
and	O
,	O
when	O
used	O
as	O
adjuvants	O
,	O
could	O
improve	O
the	O
cure	O
rates	O
achievable	O
with	O
standard	O
therapies	O
.	O

The	O
therapeutic	O
options	O
available	O
in	O
this	O
regard	O
include	O
measures	O
that	O
:	O
down	O
-	O
regulate	O
serum	B-Organism_substance
free	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
I	I-Gene_or_gene_product
;	O
suppress	O
the	O
synthesis	O
of	O
mevalonic	B-Simple_chemical
acid	I-Simple_chemical
and	O
/	O
or	O
certain	O
derivatives	O
thereof	O
;	O
modulate	O
arachidonate	B-Simple_chemical
metabolism	O
by	O
inhibiting	O
5	B-Gene_or_gene_product
-	I-Gene_or_gene_product
lipoxygenase	I-Gene_or_gene_product
,	O
12	B-Gene_or_gene_product
-	I-Gene_or_gene_product
lipoxygenase	I-Gene_or_gene_product
,	O
or	O
COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
;	O
antagonize	O
the	O
activation	O
of	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
transcription	O
factors	O
;	O
promote	O
the	O
activation	O
of	O
PPAR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
gamma	I-Gene_or_gene_product
transcription	O
factors	O
;	O
and	O
that	O
suppress	O
angiogenesis	O
by	O
additional	O
mechanisms	O
.	O

Many	O
of	O
these	O
measures	O
appear	O
suitable	O
for	O
use	O
in	O
cancer	B-Cancer
prevention	O
.	O

Oncogenic	O
mechanisms	O
of	O
Evi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
protein	O
.	O

Although	O
Evi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
is	O
thought	O
to	O
promote	O
growth	O
or	O
block	O
differentiation	O
in	O
some	O
cell	B-Cell
types	O
,	O
its	O
biological	O
functions	O
have	O
not	O
been	O
elucidated	O
.	O

To	O
explore	O
the	O
mechanisms	O
underlying	O
Evi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
induced	O
oncogenesis	O
,	O
we	O
investigated	O
whether	O
Evi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
affects	O
the	O
signaling	O
of	O
transforming	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
(	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
)	O
,	O
which	O
inhibits	O
proliferation	O
of	O
a	O
wide	O
range	O
of	O
cell	B-Cell
types	I-Cell
and	O
is	O
one	O
of	O
the	O
most	O
studied	O
growth	O
regulatory	O
factors	O
.	O

We	O
demonstrated	O
that	O
Evi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
represses	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
signaling	O
and	O
antagonizes	O
its	O
growth	O
-	O
inhibitory	O
effects	O
.	O

Two	O
separate	O
regions	O
of	O
Evi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
are	O
responsible	O
for	O
this	O
repression	O
,	O
one	O
of	O
which	O
is	O
the	O
first	O
zinc	O
-	O
finger	O
domain	O
.	O

Through	O
this	O
domain	O
,	O
Evi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
physically	O
interacts	O
with	O
Smad3	B-Gene_or_gene_product
,	O
an	O
intracellular	B-Immaterial_anatomical_entity
mediator	O
of	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
signaling	O
,	O
thereby	O
suppressing	O
the	O
transcriptional	O
activity	O
of	O
Smad3	B-Gene_or_gene_product
.	O

These	O
results	O
define	O
a	O
novel	O
function	O
of	O
Evi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
as	O
a	O
repressor	O
of	O
signaling	O
components	O
of	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
.	O

We	O
also	O
demonstrated	O
that	O
Evi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
represses	O
Smad	B-Gene_or_gene_product
-	O
induced	O
transcriptional	O
activation	O
by	O
recruiting	O
CtBP	B-Gene_or_gene_product
as	O
a	O
corepressor	O
.	O

Evi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
associates	O
with	O
CtBP1	B-Gene_or_gene_product
through	O
one	O
of	O
the	O
CtBP	B-Gene_or_gene_product
-	O
binding	O
consensus	O
motifs	O
within	O
the	O
region	O
from	O
amino	B-Amino_acid
acid	I-Amino_acid
544	I-Amino_acid
to	O
607	B-Amino_acid
,	O
and	O
this	O
association	O
is	O
required	O
for	O
the	O
efficient	O
inhibition	O
of	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
signaling	O
.	O

A	O
specific	O
histone	B-Gene_or_gene_product
deacetylase	I-Gene_or_gene_product
(	O
HDAc	B-Gene_or_gene_product
)	O
inhibitor	O
,	O
trichostatin	B-Simple_chemical
A	I-Simple_chemical
(	O
TSA	B-Simple_chemical
)	O
,	O
alleviates	O
Evi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
mediated	O
repression	O
of	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
signaling	O
,	O
suggesting	O
that	O
HDAc	B-Gene_or_gene_product
is	O
involved	O
in	O
transcriptional	O
repression	O
by	O
Evi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

This	O
identifies	O
a	O
novel	O
function	O
of	O
Evi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
as	O
a	O
member	O
of	O
corepressor	O
complexes	O
and	O
suggests	O
that	O
aberrant	O
recruitment	O
of	O
corepressors	O
is	O
one	O
of	O
the	O
mechanisms	O
involved	O
in	O
Evi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
induced	O
leukemogenesis	O
.	O

These	O
results	O
indicate	O
that	O
specific	O
HDAc	B-Gene_or_gene_product
inhibitors	O
may	O
be	O
useful	O
in	O
the	O
treatment	O
of	O
Evi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
induced	O
neoplastic	B-Cancer
tumors	I-Cancer
,	O
including	O
myeloid	B-Cancer
leukemias	I-Cancer
.	O

Antibodies	O
to	O
PAI	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
alter	O
the	O
invasive	O
and	O
migratory	O
properties	O
of	O
human	B-Organism
tumour	B-Cell
cells	I-Cell
in	O
vitro	O
.	O

Recent	O
reports	O
suggest	O
that	O
elevated	O
levels	O
of	O
plasminogen	B-Gene_or_gene_product
activator	I-Gene_or_gene_product
inhibitor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
PAI	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
may	O
contribute	O
to	O
tumour	B-Cancer
progression	O
.	O

The	O
studies	O
reported	O
here	O
were	O
designed	O
to	O
help	O
elucidate	O
PAI	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
'	O
s	O
contribution	O
to	O
the	O
invasive	O
and	O
migratory	O
phenotype	O
.	O

Antibodies	O
to	O
PA	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
dose	O
-	O
dependently	O
,	O
and	O
significantly	O
,	O
inhibited	O
the	O
invasive	O
and	O
migratory	O
potential	O
of	O
human	B-Organism
HT1080	B-Cell
fibrosarcoma	I-Cell
cells	I-Cell
,	O
as	O
did	O
an	O
antibody	O
to	O
uPA	B-Gene_or_gene_product
and	O
the	O
plasmin	B-Gene_or_gene_product
inhibitor	O
aprotinin	B-Gene_or_gene_product
.	O

Invasion	O
of	O
the	O
human	B-Organism
melanoma	B-Cell
cell	I-Cell
line	I-Cell
,	O
BLM	B-Cell
,	O
was	O
also	O
attenuated	O
by	O
the	O
anti	O
-	O
PAI	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
monoclonal	O
antibody	O
MAI	O
-	O
12	O
.	O

The	O
non	O
-	O
invasive	O
human	B-Organism
melanoma	B-Cell
cell	I-Cell
line	I-Cell
,	O
IF6	B-Cell
,	O
which	O
does	O
not	O
express	O
uPA	B-Gene_or_gene_product
,	O
provided	O
further	O
confirmation	O
of	O
PAI	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
uPA	B-Gene_or_gene_product
'	O
s	O
role	O
as	O
,	O
upon	O
transfection	O
with	O
uPA	B-Gene_or_gene_product
,	O
this	O
cell	B-Cell
line	I-Cell
attained	O
an	O
invasive	O
phenotype	O
,	O
which	O
was	O
again	O
attenuated	O
by	O
MAI	O
-	O
12	O
.	O

Although	O
antibodies	O
to	O
PAI	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
did	O
not	O
affect	O
the	O
adhesion	O
of	O
HT1080	B-Cell
cells	I-Cell
to	O
vitronectin	B-Gene_or_gene_product
,	O
the	O
antibody	O
to	O
uPA	B-Gene_or_gene_product
reduced	O
their	O
attachment	O
.	O

Addition	O
of	O
exogenous	O
PAI	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
however	O
,	O
prevented	O
HT1080	B-Cell
cell	I-Cell
adhesion	O
(	O
IC50	O
180	O
nM	O
)	O
and	O
promoted	O
cell	B-Cell
detachment	O
from	O
vitronectin	B-Gene_or_gene_product
.	O

Furthermore	O
melanoma	B-Cell
cells	I-Cell
transfected	O
with	O
a	O
uPA	B-Gene_or_gene_product
variant	O
,	O
which	O
had	O
an	O
impaired	O
interaction	O
with	O
PAI	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
were	O
not	O
invasive	O
and	O
had	O
impaired	O
binding	O
to	O
vitronectin	B-Gene_or_gene_product
.	O

These	O
data	O
highlight	O
the	O
importance	O
of	O
a	O
balanced	O
proteolysis	O
and	O
suggest	O
an	O
additional	O
role	O
for	O
PAI	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
distinct	O
from	O
its	O
role	O
in	O
proteolysis	O
.	O

These	O
data	O
also	O
suggest	O
that	O
uPA	B-Gene_or_gene_product
and	O
PAI	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
may	O
co	O
-	O
operate	O
in	O
the	O
migratory	O
process	O
by	O
respectively	O
facilitating	O
the	O
attachment	O
to	O
,	O
and	O
subsequent	O
detachment	O
from	O
,	O
vitronectin	B-Gene_or_gene_product
in	O
the	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
.	O

These	O
results	O
support	O
the	O
clinical	O
findings	O
and	O
indicate	O
that	O
modulation	O
of	O
PAI	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
activity	O
may	O
be	O
of	O
therapeutic	O
benefit	O
for	O
the	O
treatment	O
of	O
cancer	B-Cancer
.	O

Inhibition	O
of	O
PDGF	B-Gene_or_gene_product
-	O
stimulated	O
and	O
matrix	B-Cellular_component
-	O
mediated	O
proliferation	O
of	O
human	B-Organism
vascular	B-Cell
smooth	I-Cell
muscle	I-Cell
cells	I-Cell
by	O
SPARC	B-Gene_or_gene_product
is	O
independent	O
of	O
changes	O
in	O
cell	B-Cell
shape	O
or	O
cyclin	B-Simple_chemical
-	I-Simple_chemical
dependent	I-Simple_chemical
kinase	I-Simple_chemical
inhibitors	I-Simple_chemical
.	O

Interactions	O
among	O
growth	O
factors	O
,	O
cells	B-Cell
,	O
and	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
regulate	O
proliferation	O
during	O
normal	O
development	O
and	O
in	O
pathologies	O
such	O
as	O
atherosclerosis	O
.	O

SPARC	B-Gene_or_gene_product
(	O
secreted	B-Gene_or_gene_product
protein	I-Gene_or_gene_product
,	I-Gene_or_gene_product
acidic	I-Gene_or_gene_product
,	I-Gene_or_gene_product
and	I-Gene_or_gene_product
rich	I-Gene_or_gene_product
in	I-Gene_or_gene_product
cysteine	I-Gene_or_gene_product
)	O
is	O
a	O
matrix	B-Cellular_component
-	O
associated	O
glycoprotein	O
that	O
modulates	O
the	O
adhesion	O
and	O
proliferation	O
of	O
vascular	B-Cell
cells	I-Cell
.	O

In	O
this	O
study	O
,	O
we	O
demonstrate	O
that	O
SPARC	B-Gene_or_gene_product
inhibits	O
human	B-Organism
arterial	B-Cell
smooth	I-Cell
muscle	I-Cell
cell	I-Cell
proliferation	O
stimulated	O
by	O
platelet	B-Gene_or_gene_product
-	I-Gene_or_gene_product
derived	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
or	O
by	O
adhesion	O
to	O
monomeric	B-Gene_or_gene_product
type	I-Gene_or_gene_product
I	I-Gene_or_gene_product
collagen	I-Gene_or_gene_product
.	O

Binding	O
studies	O
with	O
SPARC	B-Gene_or_gene_product
and	O
SPARC	B-Gene_or_gene_product
peptides	I-Gene_or_gene_product
indicate	O
specific	O
and	O
saturable	O
interaction	O
with	O
smooth	B-Cell
muscle	I-Cell
cells	I-Cell
that	O
involves	O
the	O
C	O
-	O
terminal	O
Ca2	O
+	O
-	O
binding	O
region	O
of	O
the	O
protein	O
.	O

We	O
also	O
report	O
that	O
SPARC	B-Gene_or_gene_product
arrests	O
monomeric	O
collagen	B-Gene_or_gene_product
-	O
supported	O
smooth	B-Cell
muscle	I-Cell
cell	I-Cell
proliferation	O
in	O
the	O
late	O
G1	O
-	O
phase	O
of	O
the	O
cell	B-Cell
cycle	O
in	O
the	O
absence	O
of	O
an	O
effect	O
on	O
cell	B-Cell
shape	O
or	O
on	O
levels	O
of	O
cyclin	B-Simple_chemical
-	I-Simple_chemical
dependent	I-Simple_chemical
kinase	I-Simple_chemical
inhibitors	I-Simple_chemical
.	O

Cyclin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
dependent	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
activity	O
,	O
p107	B-Gene_or_gene_product
and	O
cyclin	B-Gene_or_gene_product
A	I-Gene_or_gene_product
levels	O
,	O
and	O
retinoblastoma	B-Gene_or_gene_product
protein	I-Gene_or_gene_product
phosphorylation	O
are	O
markedly	O
reduced	O
in	O
response	O
to	O
the	O
addition	O
of	O
exogenous	O
SPARC	B-Gene_or_gene_product
and	O
/	O
or	O
peptides	O
derived	O
from	O
specific	O
domains	O
of	O
SPARC	B-Gene_or_gene_product
.	O

Thus	O
,	O
SPARC	B-Gene_or_gene_product
,	O
previously	O
characterized	O
as	O
an	O
inhibitor	O
of	O
platelet	B-Gene_or_gene_product
-	I-Gene_or_gene_product
derived	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
binding	O
to	O
its	O
receptor	O
,	O
also	O
antagonizes	O
smooth	B-Cell
muscle	I-Cell
cell	I-Cell
proliferation	O
mediated	O
by	O
monomeric	O
collagen	B-Gene_or_gene_product
at	O
the	O
level	O
of	O
cyclin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
dependent	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
activity	O
.	O

Prognostic	O
significance	O
of	O
heat	B-Gene_or_gene_product
shock	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
27	I-Gene_or_gene_product
(	O
HSP27	B-Gene_or_gene_product
)	O
in	O
patients	B-Organism
with	O
oral	B-Cancer
squamous	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
.	O

Heat	B-Gene_or_gene_product
shock	I-Gene_or_gene_product
proteins	I-Gene_or_gene_product
(	O
HSPs	B-Gene_or_gene_product
)	O
have	O
been	O
defined	O
as	O
proteins	O
induced	O
by	O
heat	O
shock	O
and	O
other	O
environmental	O
and	O
pathophysiologic	O
stress	O
.	O

Heat	B-Gene_or_gene_product
shock	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
27	I-Gene_or_gene_product
(	O
HSP27	B-Gene_or_gene_product
)	O
is	O
one	O
of	O
the	O
small	B-Gene_or_gene_product
heat	I-Gene_or_gene_product
shock	I-Gene_or_gene_product
proteins	I-Gene_or_gene_product
.	O

HSP27	B-Gene_or_gene_product
is	O
implicated	O
in	O
protein	O
-	O
protein	O
interactions	O
such	O
as	O
folding	O
,	O
translocation	O
,	O
and	O
prevention	O
of	O
inappropriate	O
protein	O
aggregation	O
.	O

Many	O
of	O
their	O
functions	O
suggest	O
that	O
they	O
play	O
important	O
roles	O
in	O
cancers	B-Cancer
.	O

Archival	B-Tissue
tissues	I-Tissue
from	O
40	O
patients	B-Organism
with	O
oral	B-Cancer
squamous	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
who	O
received	O
primary	O
surgical	O
resection	O
were	O
examined	O
for	O
HSP27	B-Gene_or_gene_product
by	O
immunohistochemistry	O
and	O
correlated	O
with	O
clinical	O
stage	O
,	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
,	O
histological	O
grade	O
and	O
survival	O
period	O
.	O

HSP27	B-Gene_or_gene_product
expression	O
was	O
positive	O
staining	O
(	O
+	O
)	O
in	O
20	O
(	O
50	O
%	O
)	O
,	O
weak	O
or	O
negative	O
staining	O
(	O
-	O
)	O
in	O
20	O
(	O
50	O
%	O
)	O
of	O
total	O
40	O
cases	O
.	O

There	O
was	O
no	O
correlation	O
between	O
HSP27	B-Gene_or_gene_product
expression	O
and	O
clinical	O
stage	O
,	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
and	O
histological	O
grade	O
.	O

However	O
,	O
when	O
compared	O
with	O
clinicopathological	O
features	O
,	O
the	O
expression	O
of	O
HSP27	B-Gene_or_gene_product
correlated	O
inversely	O
with	O
survival	O
period	O
.	O

This	O
study	O
suggests	O
that	O
the	O
expression	O
of	O
HSP27	B-Gene_or_gene_product
is	O
frequently	O
promoted	O
in	O
patients	B-Organism
with	O
oral	B-Cancer
squamous	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
and	O
should	O
be	O
considered	O
an	O
independent	O
prognostic	O
factor	O
of	O
oral	B-Cancer
squamous	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
patients	B-Organism
.	O

Upregulation	O
of	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
receptors	I-Gene_or_gene_product
is	O
associated	O
with	O
advanced	O
neuroblastoma	B-Cancer
.	O

BACKGROUND	O
:	O
Angiogenesis	O
is	O
essential	O
for	O
tumor	B-Cancer
growth	O
and	O
relies	O
on	O
the	O
production	O
of	O
angiogenic	O
factors	O
.	O

Vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
is	O
a	O
major	O
regulator	O
of	O
angiogenesis	O
that	O
binds	O
to	O
tyrosine	O
kinase	O
receptors	O
Flt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
KDR	B-Gene_or_gene_product
.	O

The	O
interaction	O
of	O
VEGF	B-Gene_or_gene_product
and	O
its	O
receptors	O
at	O
gene	O
and	O
protein	O
levels	O
in	O
neuroblastoma	B-Cancer
remains	O
widely	O
unknown	O
.	O

METHODS	O
:	O
Tumor	B-Cancer
biopsy	I-Cancer
specimens	I-Cancer
and	O
serum	B-Organism_substance
were	O
obtained	O
from	O
37	O
neuroblastoma	B-Cancer
patients	B-Organism
;	O
adrenal	B-Multi-tissue_structure
biopsy	I-Multi-tissue_structure
sections	I-Multi-tissue_structure
and	O
sera	B-Organism_substance
of	O
7	O
normal	O
children	B-Organism
served	O
as	O
controls	O
.	O

Biopsy	B-Cancer
specimens	I-Cancer
were	O
examined	O
by	O
real	O
-	O
time	O
reverse	O
transcription	O
polymerase	O
chain	O
reaction	O
(	O
RT	O
-	O
PCR	O
)	O
and	O
Western	O
blotting	O
;	O
serum	B-Organism_substance
was	O
analyzed	O
by	O
enzyme	O
-	O
linked	O
immunosorbent	O
assay	O
(	O
ELISA	O
)	O
.	O

VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
A	I-Gene_or_gene_product
(	I-Gene_or_gene_product
165	I-Gene_or_gene_product
)	I-Gene_or_gene_product
,	O
B	B-Gene_or_gene_product
,	O
C	B-Gene_or_gene_product
,	O
Flt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
and	O
KDR	B-Gene_or_gene_product
were	O
analyzed	O
.	O

RESULTS	O
:	O
VEGF	B-Gene_or_gene_product
isoforms	O
and	O
its	O
receptors	O
'	O
mRNA	O
were	O
expressed	O
in	O
neuroblastoma	B-Cancer
and	O
control	O
tissues	B-Tissue
.	O

Whereas	O
the	O
ligands	O
were	O
increased	O
in	O
stages	O
III	O
and	O
IV	O
,	O
the	O
receptors	O
were	O
upregulated	O
in	O
stage	O
III	O
only	O
.	O

At	O
protein	O
level	O
,	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
B	I-Gene_or_gene_product
and	O
C	B-Gene_or_gene_product
,	O
Flt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
and	O
KDR	B-Gene_or_gene_product
were	O
not	O
detectable	O
in	O
tissue	B-Organism_substance
lysates	I-Organism_substance
,	O
whereas	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
A	I-Gene_or_gene_product
was	O
increased	O
in	O
stages	O
III	O
and	O
IV	O
.	O

Serum	B-Organism_substance
VEGF	B-Gene_or_gene_product
protein	O
levels	O
were	O
upregulated	O
in	O
stage	O
III	O
.	O

CONCLUSIONS	O
:	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
A	I-Gene_or_gene_product
(	I-Gene_or_gene_product
165	I-Gene_or_gene_product
)	I-Gene_or_gene_product
is	O
one	O
of	O
the	O
major	O
angiogenesis	O
regulators	O
among	O
the	O
ligands	O
'	O
family	O
of	O
VEGF	B-Gene_or_gene_product
,	O
whereas	O
its	O
receptors	O
KDR	B-Gene_or_gene_product
,	O
and	O
most	O
probably	O
Flt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
may	O
contribute	O
to	O
a	O
poor	O
prognosis	O
(	O
angiogenic	O
)	O
phenotype	O
,	O
as	O
indicated	O
by	O
their	O
upregulated	O
MRNA	O
levels	O
in	O
stage	B-Cancer
III	I-Cancer
neuroblastoma	I-Cancer
.	O

VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
A	I-Gene_or_gene_product
(	I-Gene_or_gene_product
165	I-Gene_or_gene_product
)	I-Gene_or_gene_product
mainly	O
contributes	O
to	O
increased	O
serum	B-Organism_substance
VEGF	B-Gene_or_gene_product
levels	O
and	O
may	O
serve	O
as	O
a	O
diagnostic	O
tool	O
in	O
advanced	B-Cancer
-	I-Cancer
stage	I-Cancer
neuroblastoma	I-Cancer
.	O

Hypoxia	B-Gene_or_gene_product
-	I-Gene_or_gene_product
inducible	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
1	I-Gene_or_gene_product
activation	O
by	O
aerobic	O
glycolysis	O
implicates	O
the	O
Warburg	O
effect	O
in	O
carcinogenesis	O
.	O

Cancer	B-Cell
cells	I-Cell
display	O
high	O
rates	O
of	O
aerobic	O
glycolysis	O
,	O
a	O
phenomenon	O
known	O
historically	O
as	O
the	O
Warburg	O
effect	O
.	O

Lactate	B-Simple_chemical
and	O
pyruvate	B-Simple_chemical
,	O
the	O
end	O
products	O
of	O
glycolysis	O
,	O
are	O
highly	O
produced	O
by	O
cancer	B-Cell
cells	I-Cell
even	O
in	O
the	O
presence	O
of	O
oxygen	B-Simple_chemical
.	O

Hypoxia	O
-	O
induced	O
gene	O
expression	O
in	O
cancer	B-Cell
cells	I-Cell
has	O
been	O
linked	O
to	O
malignant	O
transformation	O
.	O

Here	O
we	O
provide	O
evidence	O
that	O
lactate	B-Simple_chemical
and	O
pyruvate	B-Simple_chemical
regulate	O
hypoxia	O
-	O
inducible	O
gene	O
expression	O
independently	O
of	O
hypoxia	O
by	O
stimulating	O
the	O
accumulation	O
of	O
hypoxia	B-Gene_or_gene_product
-	I-Gene_or_gene_product
inducible	I-Gene_or_gene_product
Factor	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
(	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
)	O
.	O

In	O
human	B-Organism
gliomas	B-Cancer
and	O
other	O
cancer	B-Cell
cell	I-Cell
lines	I-Cell
,	O
the	O
accumulation	O
of	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
protein	O
under	O
aerobic	O
conditions	O
requires	O
the	O
metabolism	O
of	O
glucose	B-Simple_chemical
to	O
pyruvate	B-Simple_chemical
that	O
prevents	O
the	O
aerobic	O
degradation	O
of	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
protein	O
,	O
activates	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
DNA	B-Cellular_component
binding	O
activity	O
,	O
and	O
enhances	O
the	O
expression	O
of	O
several	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
activated	O
genes	O
including	O
erythropoietin	B-Gene_or_gene_product
,	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
,	O
glucose	B-Gene_or_gene_product
transporter	I-Gene_or_gene_product
3	I-Gene_or_gene_product
,	O
and	O
aldolase	B-Gene_or_gene_product
A	I-Gene_or_gene_product
.	O

Our	O
findings	O
support	O
a	O
novel	O
role	O
for	O
pyruvate	B-Simple_chemical
in	O
metabolic	O
signaling	O
and	O
suggest	O
a	O
mechanism	O
by	O
which	O
high	O
rates	O
of	O
aerobic	O
glycolysis	O
can	O
promote	O
the	O
malignant	O
transformation	O
and	O
survival	O
of	O
cancer	B-Cell
cells	I-Cell
.	O

Risk	O
of	O
myelodysplastic	O
syndrome	O
and	O
acute	B-Cancer
myeloid	I-Cancer
leukemia	I-Cancer
in	O
congenital	O
neutropenias	O
.	O

Granulocyte	B-Gene_or_gene_product
colony	I-Gene_or_gene_product
-	I-Gene_or_gene_product
stimulating	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
G	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
)	O
has	O
had	O
a	O
major	O
impact	O
on	O
the	O
management	O
of	O
"	O
severe	O
chronic	O
neutropenia	O
"	O
(	O
SCN	O
)	O
,	O
a	O
collective	O
term	O
referring	O
to	O
congenital	O
,	O
idiopathic	O
,	O
or	O
cyclic	O
neutropenia	O
.	O

Almost	O
all	O
patients	B-Organism
respond	O
to	O
G	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
with	O
increased	O
neutrophils	B-Cell
,	O
reduced	O
infections	O
,	O
and	O
improved	O
survival	O
.	O

Some	O
responders	O
with	O
congenital	O
neutropenia	O
and	O
Shwachman	O
-	O
Diamond	O
syndrome	O
(	O
SDS	O
)	O
have	O
developed	O
myelodysplastic	O
syndrome	O
and	O
acute	B-Cancer
myeloid	I-Cancer
leukemia	I-Cancer
(	O
MDS	O
/	O
AML	B-Cancer
)	O
,	O
which	O
raises	O
the	O
question	O
of	O
the	O
role	O
of	O
G	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
in	O
pathogenesis	O
.	O

The	O
issue	O
is	O
complicated	O
because	O
both	O
disorders	O
have	O
a	O
propensity	O
for	O
MDS	O
or	O
AML	B-Cancer
as	O
part	O
of	O
their	O
natural	O
history	O
.	O

To	O
address	O
this	O
,	O
the	O
Severe	O
Chronic	O
Neutropenia	O
International	O
Registry	O
(	O
SCNIR	O
)	O
used	O
its	O
large	O
database	O
of	O
chronic	O
neutropenia	O
patients	B-Organism
treated	O
with	O
G	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
to	O
determine	O
the	O
incidence	O
of	O
malignant	B-Cell
myeloid	I-Cell
transformation	O
in	O
the	O
two	O
disorders	O
,	O
and	O
its	O
relationship	O
to	O
treatment	O
and	O
to	O
other	O
patient	B-Organism
characteristics	O
.	O

No	O
statistically	O
significant	O
relationships	O
were	O
found	O
between	O
age	O
at	O
onset	O
of	O
MDS	O
or	O
AML	B-Cancer
and	O
patient	B-Organism
gender	O
,	O
G	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
dose	O
,	O
or	O
duration	O
of	O
G	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
therapy	O
.	O

What	O
was	O
observed	O
,	O
however	O
,	O
was	O
the	O
multistep	O
acquisition	O
of	O
aberrant	O
cellular	B-Cell
genetic	O
changes	O
in	O
marrow	B-Cell
cells	I-Cell
from	O
patients	B-Organism
who	O
transformed	O
,	O
including	O
activating	O
ras	B-Gene_or_gene_product
oncogene	O
mutations	O
,	O
clonal	O
cytogenetic	O
abnormalities	O
,	O
and	O
G	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
mutations	O
.	O

In	O
murine	B-Organism
models	O
,	O
the	O
latter	O
produces	O
a	O
hyperproliferative	O
response	O
to	O
G	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
,	O
confers	O
resistance	O
to	O
apoptosis	O
,	O
and	O
enhances	O
cell	B-Cell
survival	O
.	O

Since	O
congenital	O
neutropenia	O
and	O
SDS	O
are	O
inherited	O
forms	O
of	O
bone	B-Multi-tissue_structure
marrow	I-Multi-tissue_structure
failure	O
,	O
G	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
may	O
accelerate	O
the	O
propensity	O
for	O
MDS	O
/	O
AML	B-Cancer
in	O
the	O
genetically	O
altered	O
stem	B-Cell
and	O
progenitor	B-Cell
cells	I-Cell
,	O
especially	O
in	O
those	O
with	O
G	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
and	O
ras	B-Gene_or_gene_product
mutations	O
(	O
82	O
%	O
and	O
50	O
%	O
of	O
patients	B-Organism
who	O
transform	O
,	O
respectively	O
)	O
.	O

Alternatively	O
,	O
and	O
equally	O
plausible	O
,	O
G	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
may	O
simply	O
be	O
an	O
"	O
innocent	O
bystander	O
"	O
that	O
corrects	O
neutropenia	O
,	O
prolongs	O
patient	B-Organism
survival	O
,	O
and	O
allows	O
time	O
for	O
the	O
malignant	O
predisposition	O
to	O
declare	O
itself	O
.	O

In	O
patients	B-Organism
who	O
transform	O
to	O
overt	O
MDS	O
or	O
AML	B-Cancer
,	O
hematopoietic	B-Cell
stem	I-Cell
cell	I-Cell
transplantation	O
is	O
the	O
only	O
chance	O
for	O
cure	O
.	O

In	O
those	O
with	O
"	O
soft	O
"	O
signs	O
of	O
MDS	O
,	O
such	O
as	O
an	O
isolated	O
clonal	O
cytogenetic	O
change	O
but	O
without	O
other	O
evidence	O
of	O
MDS	O
,	O
or	O
with	O
an	O
isolated	O
G	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
mutation	O
,	O
there	O
is	O
room	O
for	O
conservative	O
management	O
.	O

One	O
option	O
is	O
to	O
reduce	O
the	O
G	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
dosage	O
as	O
much	O
as	O
possible	O
,	O
and	O
observe	O
the	O
tempo	O
of	O
progression	O
,	O
if	O
any	O
,	O
to	O
more	O
overt	O
signs	O
of	O
malignancy	B-Cancer
.	O

Hypoxic	O
induction	O
of	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
and	O
VEGF	B-Gene_or_gene_product
expression	O
in	O
head	B-Cell
and	I-Cell
neck	I-Cell
squamous	I-Cell
cell	I-Cell
carcinoma	I-Cell
lines	I-Cell
is	O
mediated	O
by	O
stress	O
activated	O
protein	O
kinases	O
.	O

Solid	B-Cancer
tumors	I-Cancer
must	O
establish	O
a	O
blood	B-Organism_substance
supply	O
in	O
order	O
to	O
proliferate	O
and	O
grow	O
.	O

Cancer	B-Cell
cells	I-Cell
secrete	O
soluble	O
factors	O
which	O
can	O
induce	O
proliferation	O
and	O
migration	O
of	O
capillary	B-Cell
endothelial	I-Cell
cells	I-Cell
.	O

Among	O
the	O
most	O
potent	O
of	O
the	O
angiogenic	O
factors	O
is	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
.	O

Increased	O
VEGF	B-Gene_or_gene_product
expression	O
by	O
malignant	B-Cancer
tumors	I-Cancer
has	O
been	O
associated	O
with	O
high	O
vascularity	O
,	O
increased	O
cancer	B-Cell
cell	I-Cell
growth	O
,	O
and	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
.	O

Reduced	O
oxygen	B-Simple_chemical
tension	O
has	O
been	O
shown	O
to	O
increase	O
VEGF	B-Gene_or_gene_product
production	O
by	O
induction	O
of	O
the	O
transcription	O
factor	O
hypoxia	B-Gene_or_gene_product
inducible	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
1	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
(	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
)	O
.	O

The	O
mechanisms	O
by	O
which	O
hypoxic	O
tumor	B-Cancer
environments	O
induce	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
and	O
VEGF	B-Gene_or_gene_product
expression	O
are	O
largely	O
unknown	O
.	O

Jun	B-Gene_or_gene_product
N	I-Gene_or_gene_product
terminal	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
(	O
JNK1	B-Gene_or_gene_product
)	O
and	O
p38	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
are	O
activated	O
by	O
a	O
variety	O
of	O
stress	O
stimuli	O
.	O

To	O
determine	O
if	O
hypoxic	O
activation	O
of	O
these	O
stress	O
activated	O
protein	O
kinases	O
regulated	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
and	O
VEGF	B-Gene_or_gene_product
expression	O
,	O
we	O
assayed	O
JNK1	B-Gene_or_gene_product
and	O
p38	B-Gene_or_gene_product
activity	O
in	O
squamous	B-Cell
cell	I-Cell
carcinoma	I-Cell
(	I-Cell
SCC	I-Cell
)	I-Cell
lines	I-Cell
grown	O
under	O
normoxic	O
or	O
hypoxic	O
conditions	O
.	O

Hypoxia	O
rapidly	O
induced	O
both	O
JNK1	B-Gene_or_gene_product
and	O
p38	B-Gene_or_gene_product
activity	O
in	O
these	O
cells	B-Cell
.	O

This	O
activation	O
correlated	O
with	O
induction	O
of	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
expression	O
and	O
DNA	B-Cellular_component
binding	O
activity	O
which	O
was	O
blocked	O
by	O
the	O
p38	B-Gene_or_gene_product
inhibitor	O
SB203580	B-Simple_chemical
.	O

Hypoxia	O
also	O
increased	O
VEGF	B-Gene_or_gene_product
production	O
by	O
SCC	B-Cell
lines	I-Cell
,	O
which	O
was	O
inhibited	O
by	O
treatment	O
with	O
SB203580	B-Simple_chemical
.	O

Overexpression	O
of	O
JNK1	B-Gene_or_gene_product
or	O
p38	B-Gene_or_gene_product
was	O
sufficient	O
to	O
induce	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
and	O
VEGF	B-Gene_or_gene_product
expression	O
.	O

These	O
results	O
indicate	O
that	O
induction	O
of	O
SAPKs	B-Gene_or_gene_product
by	O
hypoxia	O
regulates	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
and	O
VEGF	B-Gene_or_gene_product
expression	O
in	O
head	B-Cell
and	I-Cell
neck	I-Cell
carcinoma	I-Cell
cell	I-Cell
lines	I-Cell
.	O

Simian	B-Organism
virus	I-Organism
40	I-Organism
large	B-Gene_or_gene_product
T	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
and	O
two	O
independent	O
T	B-Gene_or_gene_product
-	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
segments	O
sensitize	O
cells	B-Cell
to	O
apoptosis	O
following	O
genotoxic	O
damage	O
.	O

The	O
simian	B-Organism
virus	I-Organism
40	I-Organism
(	O
SV40	B-Organism
)	O
large	B-Gene_or_gene_product
tumor	I-Gene_or_gene_product
(	I-Gene_or_gene_product
T	I-Gene_or_gene_product
)	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
is	O
sufficient	O
to	O
transform	O
cells	B-Cell
in	O
cultures	O
and	O
induce	O
tumors	B-Cancer
in	O
experimental	O
animals	O
.	O

Transformation	O
of	O
primary	O
cells	B-Cell
in	O
cultures	O
requires	O
both	O
overcoming	O
growth	O
arrest	O
by	O
stimulating	O
the	O
cell	B-Cell
cycle	O
and	O
blocking	O
cell	B-Cell
death	O
activities	O
presumably	O
activated	O
by	O
oncogene	O
-	O
mediated	O
hyperproliferation	O
signals	O
.	O

The	O
study	O
presented	O
here	O
examined	O
the	O
ability	O
of	O
specific	O
regions	O
and	O
activities	O
of	O
T	B-Gene_or_gene_product
antigen	I-Gene_or_gene_product
to	O
modulate	O
apoptosis	O
in	O
cells	B-Cell
treated	O
with	O
the	O
genotoxic	O
agent	O
5	B-Simple_chemical
-	I-Simple_chemical
fluorouracil	I-Simple_chemical
(	O
5	B-Simple_chemical
-	I-Simple_chemical
FU	I-Simple_chemical
)	O
.	O

The	O
results	O
showed	O
that	O
the	O
expression	O
of	O
full	O
-	O
length	O
T	B-Gene_or_gene_product
antigen	I-Gene_or_gene_product
rendered	O
rat	B-Organism
embryo	B-Cell
fibroblasts	I-Cell
(	O
REF	B-Cell
)	O
sensitive	O
to	O
5	B-Simple_chemical
-	I-Simple_chemical
FU	I-Simple_chemical
-	O
induced	O
apoptosis	O
.	O

Thus	O
,	O
neither	O
the	O
p53	B-Gene_or_gene_product
-	O
binding	O
region	O
nor	O
the	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
homology	O
region	O
of	O
T	B-Gene_or_gene_product
antigen	I-Gene_or_gene_product
was	O
sufficient	O
to	O
prevent	O
cell	B-Cell
death	O
induced	O
by	O
the	O
DNA	B-Cellular_component
-	O
damaging	O
agent	O
.	O

T	B-Gene_or_gene_product
-	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
-	O
mediated	O
sensitization	O
occurred	O
independently	O
of	O
retinoblastoma	B-Gene_or_gene_product
protein	I-Gene_or_gene_product
or	O
p53	B-Gene_or_gene_product
and	O
p300	B-Gene_or_gene_product
binding	O
.	O

An	O
N	O
-	O
terminal	O
segment	O
containing	O
the	O
first	O
127	O
T	B-Gene_or_gene_product
-	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
amino	B-Amino_acid
acids	I-Amino_acid
(	O
T1	O
-	O
127	O
)	O
was	O
sufficient	O
to	O
sensitize	O
cells	B-Cell
.	O

A	O
C	O
-	O
terminal	O
segment	O
consisting	O
of	O
T	B-Gene_or_gene_product
-	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
amino	B-Amino_acid
acids	I-Amino_acid
251	O
to	O
708	O
(	O
T251	O
-	O
708	O
)	O
also	O
sensitized	O
cells	B-Cell
to	O
5	B-Simple_chemical
-	I-Simple_chemical
FU	I-Simple_chemical
-	O
induced	O
apoptosis	O
.	O

This	O
sensitization	O
did	O
not	O
occur	O
when	O
T251	O
-	O
708	O
was	O
targeted	O
to	O
the	O
nucleus	B-Cellular_component
by	O
inclusion	O
of	O
the	O
SV40	B-Organism
nuclear	O
localization	O
signal	O
.	O

The	O
introduction	O
of	O
mutations	O
into	O
the	O
T	B-Gene_or_gene_product
-	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
J	O
domain	O
resulted	O
in	O
mutation	O
-	O
specific	O
and	O
variable	O
inhibition	O
of	O
apoptosis	O
.	O

This	O
result	O
suggested	O
that	O
either	O
the	O
structural	O
or	O
the	O
functional	O
integrity	O
of	O
the	O
J	O
domain	O
is	O
required	O
to	O
sensitize	O
cells	B-Cell
to	O
apoptosis	O
.	O

Treatment	O
of	O
REF	B-Cell
or	O
REF	B-Cell
expressing	O
full	O
-	O
length	O
T	B-Gene_or_gene_product
antigen	I-Gene_or_gene_product
,	O
an	O
N	O
-	O
terminal	O
segment	O
,	O
or	O
T251	O
-	O
708	O
resulted	O
in	O
increased	O
expression	O
of	O
the	O
p53	B-Gene_or_gene_product
-	O
responsive	O
MDM2	B-Gene_or_gene_product
gene	O
;	O
apoptosis	O
occurred	O
through	O
a	O
p53	B-Gene_or_gene_product
-	O
dependent	O
pathway	O
,	O
as	O
p53	B-Gene_or_gene_product
-	O
null	O
cells	O
expressing	O
these	O
T	B-Gene_or_gene_product
antigens	I-Gene_or_gene_product
were	O
resistant	O
to	O
5	B-Simple_chemical
-	I-Simple_chemical
FU	I-Simple_chemical
-	O
induced	O
apoptosis	O
.	O

Possible	O
mechanisms	O
involved	O
in	O
sensitizing	O
cells	B-Cell
to	O
a	O
p53	B-Gene_or_gene_product
-	O
dependent	O
apoptosis	O
pathway	O
in	O
spite	O
of	O
the	O
ability	O
of	O
T	B-Gene_or_gene_product
antigen	I-Gene_or_gene_product
to	O
bind	O
and	O
inactivate	O
the	O
transcriptional	O
transactivating	O
activity	O
of	O
p53	B-Gene_or_gene_product
are	O
discussed	O
.	O

Optimizing	O
treatment	O
of	O
choroidal	B-Multi-tissue_structure
neovascularization	O
feeder	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
associated	O
with	O
age	O
-	O
related	O
macular	B-Tissue
degeneration	O
.	O

PURPOSE	O
:	O
To	O
optimize	O
the	O
method	O
of	O
treating	O
choroidal	B-Multi-tissue_structure
neovascularization	O
(	O
CNV	O
)	O
associated	O
with	O
age	O
-	O
related	O
macular	B-Tissue
degeneration	O
(	O
AMD	O
)	O
.	O

DESIGN	O
:	O
Experimental	O
study	O
and	O
interventional	O
case	O
series	O
.	O

METHODS	O
:	O
The	O
parameters	O
associated	O
with	O
locating	O
and	O
then	O
photocoagulating	O
CNV	O
feeder	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
were	O
identified	O
and	O
optimized	O
using	O
published	O
data	O
and	O
data	O
derived	O
from	O
modeling	O
the	O
choroidal	B-Multi-tissue_structure
vasculature	I-Multi-tissue_structure
.	O

Based	O
on	O
these	O
optimized	O
parameters	O
,	O
a	O
prototype	O
diagnostic	O
/	O
treatment	O
system	O
was	O
designed	O
that	O
captures	O
high	O
-	O
speed	O
indocyanine	B-Simple_chemical
green	I-Simple_chemical
(	O
ICG	B-Simple_chemical
)	O
angiogram	O
images	O
and	O
facilitates	O
analysis	O
of	O
the	O
images	O
by	O
enhancing	O
visualization	O
of	O
dye	O
movement	O
through	O
CNV	O
feeder	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
(	O
FVs	B-Multi-tissue_structure
)	O
.	O

The	O
system	O
also	O
permits	O
precise	O
aiming	O
and	O
delivery	O
of	O
810	O
-	O
nm	O
wavelength	O
photocoagulation	O
laser	O
energy	O
to	O
target	O
FVs	B-Multi-tissue_structure
on	O
a	O
real	O
-	O
time	O
ICG	B-Simple_chemical
angiogram	O
image	O
of	O
the	O
choroidal	B-Multi-tissue_structure
vasculature	I-Multi-tissue_structure
.	O

Target	O
FVs	B-Multi-tissue_structure
are	O
tracked	O
by	O
a	O
joy	O
-	O
stick	O
controlled	O
laser	O
aiming	O
beam	O
until	O
an	O
intravenously	B-Immaterial_anatomical_entity
-	O
injected	O
high	O
-	O
concentration	O
ICG	B-Simple_chemical
dye	I-Simple_chemical
bolus	O
is	O
observed	O
to	O
enter	O
the	O
target	O
vessel	B-Multi-tissue_structure
,	O
at	O
which	O
time	O
the	O
laser	O
is	O
fired	O
.	O

Proof	O
of	O
principle	O
of	O
the	O
combined	O
diagnosis	O
/	O
treatment	O
system	O
design	O
for	O
performing	O
dye	O
-	O
enhanced	O
photocoagulation	O
(	O
DEP	O
)	O
in	O
the	O
clinical	O
setting	O
and	O
determination	O
of	O
the	O
minimum	O
DEP	O
laser	O
energy	O
needed	O
to	O
close	O
CNV	O
FVs	B-Multi-tissue_structure
was	O
made	O
in	O
11	O
AMD	O
patients	B-Organism
requiring	O
treatment	O
of	O
CNV	O
,	O
but	O
for	O
whom	O
other	O
treatment	O
was	O
not	O
appropriate	O
.	O

RESULTS	O
:	O
Using	O
ICG	B-Simple_chemical
-	O
DEP	O
,	O
CNV	O
feeder	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
were	O
closed	O
with	O
single	O
pulse	O
laser	O
energy	O
,	O
delivering	O
as	O
little	O
as	O
0	O
.	O
6	O
to	O
1	O
.	O
8	O
J	O
of	O
energy	O
to	O
the	O
fundus	B-Multi-tissue_structure
,	O
producing	O
no	O
visible	O
change	O
in	O
the	O
fundus	B-Multi-tissue_structure
.	O

Successful	O
FV	B-Multi-tissue_structure
closure	O
was	O
usually	O
indicated	O
immediately	O
by	O
presence	O
of	O
incarcerated	O
ICG	B-Simple_chemical
dye	I-Simple_chemical
in	O
the	O
vessel	B-Multi-tissue_structure
adjacent	O
to	O
the	O
burn	O
site	O
.	O

The	O
prototype	O
system	O
proved	O
relatively	O
easy	O
to	O
operate	O
.	O

After	O
acquiring	O
and	O
interpreting	O
diagnostic	O
angiograms	O
and	O
repositioning	O
a	O
patient	B-Organism
in	O
front	O
of	O
the	O
device	O
,	O
feeder	B-Multi-tissue_structure
vessel	I-Multi-tissue_structure
DEP	O
and	O
treatment	O
evaluation	O
required	O
15	O
to	O
20	O
minutes	O
.	O

CONCLUSIONS	O
:	O
Indocyanine	B-Simple_chemical
green	I-Simple_chemical
dye	I-Simple_chemical
-	O
enhanced	O
photocoagulation	O
of	O
CNV	O
feeder	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
,	O
facilitated	O
by	O
use	O
of	O
a	O
device	O
that	O
permits	O
real	O
-	O
time	O
visualization	O
of	O
the	O
choroidal	B-Multi-tissue_structure
circulation	O
while	O
aiming	O
the	O
treatment	O
laser	O
beam	O
,	O
appears	O
to	O
minimize	O
the	O
amount	O
of	O
energy	O
applied	O
to	O
the	O
fundus	B-Multi-tissue_structure
and	O
the	O
volume	O
of	O
fundus	B-Tissue
tissue	I-Tissue
affected	O
by	O
treatment	O
,	O
compared	O
with	O
other	O
treatment	O
modalities	O
.	O

The	O
combination	O
diagnosis	O
/	O
treatment	O
device	O
should	O
be	O
useful	O
in	O
optimizing	O
FV	B-Multi-tissue_structure
treatment	O
and	O
in	O
refining	O
and	O
evaluating	O
the	O
efficacy	O
of	O
DEP	O
in	O
future	O
clinical	O
trials	O
.	O

Inhibition	O
of	O
angiogenesis	O
by	O
the	O
cancer	B-Cancer
chemopreventive	O
agent	O
conjugated	B-Simple_chemical
linoleic	I-Simple_chemical
acid	I-Simple_chemical
.	O

Dietary	O
conjugated	B-Simple_chemical
linoleic	I-Simple_chemical
acid	I-Simple_chemical
(	O
CLA	B-Simple_chemical
)	O
has	O
been	O
shown	O
previously	O
to	O
inhibit	O
rat	B-Organism
mammary	B-Organ
carcinogenesis	O
.	O

In	O
addition	O
to	O
direct	O
effects	O
on	O
mammary	B-Cell
epithelial	I-Cell
cells	I-Cell
,	O
including	O
decreased	O
proliferation	O
and	O
induction	O
of	O
apoptosis	O
,	O
CLA	B-Simple_chemical
may	O
exert	O
its	O
effects	O
indirectly	O
by	O
inhibiting	O
the	O
differentiation	O
of	O
mammary	B-Cell
stromal	I-Cell
cells	I-Cell
to	O
an	O
endothelial	B-Cell
cell	I-Cell
type	I-Cell
.	O

Specifically	O
,	O
CLA	B-Simple_chemical
was	O
found	O
to	O
decrease	O
the	O
ability	O
of	O
mammary	B-Cell
stromal	I-Cell
cells	I-Cell
to	O
form	O
complex	O
anastomosing	O
microcapillary	B-Multi-tissue_structure
networks	I-Multi-tissue_structure
in	O
vitro	O
on	O
Engelbreth	O
-	O
Holm	O
-	O
Swarm	O
-	O
derived	O
reconstituted	O
basement	B-Cellular_component
membrane	I-Cellular_component
.	O

This	O
suggested	O
that	O
CLA	B-Simple_chemical
might	O
inhibit	O
angiogenesis	O
in	O
vivo	O
.	O

To	O
test	O
this	O
possibility	O
,	O
CD2	O
/	O
F	O
(	O
1	O
)	O
mice	B-Organism
were	O
placed	O
on	O
synthetic	O
diets	O
containing	O
0	O
,	O
1	O
,	O
or	O
2	O
%	O
CLA	B-Simple_chemical
for	O
6	O
weeks	O
,	O
before	O
angiogenic	O
challenge	O
by	O
s	O
.	O
c	O
.	O
injection	O
with	O
an	O
angiogenic	O
gel	O
substrate	O
(	O
Matrigel	O
pellet	O
assay	O
)	O
.	O

After	O
7	O
days	O
,	O
the	O
pellets	O
from	O
animals	O
fed	O
the	O
control	O
diet	O
were	O
infiltrated	O
by	O
abundant	O
branching	O
networks	B-Multi-tissue_structure
of	I-Multi-tissue_structure
blood	I-Multi-tissue_structure
vessels	I-Multi-tissue_structure
with	O
patent	O
lumen	B-Immaterial_anatomical_entity
-	O
containing	O
RBCs	B-Cell
.	O

In	O
contrast	O
,	O
pellets	O
from	O
the	O
CLA	B-Simple_chemical
-	O
fed	O
animals	O
contained	O
fewer	O
infiltrating	O
cells	B-Cell
,	O
which	O
formed	O
limited	O
branching	O
cellular	B-Multi-tissue_structure
networks	I-Multi-tissue_structure
,	O
the	O
majority	O
of	O
which	O
had	O
collapsed	O
lumen	B-Immaterial_anatomical_entity
and	O
no	O
RBCs	B-Cell
.	O

Both	O
levels	O
of	O
dietary	O
CLA	B-Simple_chemical
showed	O
similar	O
effects	O
,	O
with	O
the	O
number	O
of	O
RBC	B-Cell
-	O
containing	O
vessels	B-Multi-tissue_structure
per	O
20x	O
field	O
decreased	O
to	O
a	O
third	O
of	O
that	O
seen	O
in	O
control	O
.	O

Dietary	O
CLA	B-Simple_chemical
decreased	O
serum	B-Organism_substance
levels	O
of	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
and	O
whole	O
mammary	B-Organ
gland	I-Organ
levels	O
of	O
VEGF	B-Gene_or_gene_product
and	O
its	O
receptor	O
Flk	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

Both	O
cis	B-Simple_chemical
-	I-Simple_chemical
9	I-Simple_chemical
,	O
trans	B-Simple_chemical
-	I-Simple_chemical
11	I-Simple_chemical
and	O
trans	B-Simple_chemical
-	I-Simple_chemical
10	I-Simple_chemical
,	O
cis	B-Simple_chemical
-	I-Simple_chemical
12	I-Simple_chemical
CLA	I-Simple_chemical
isomers	I-Simple_chemical
were	O
effective	O
in	O
inhibiting	O
angiogenesis	O
in	O
vitro	O
in	O
a	O
dose	O
-	O
dependent	O
fashion	O
.	O

The	O
ability	O
of	O
CLA	B-Simple_chemical
to	O
inhibit	O
angiogenesis	O
may	O
contribute	O
to	O
its	O
efficacy	O
as	O
a	O
chemopreventive	O
agent	O
.	O

Molecular	O
characterization	O
of	O
angiogenic	O
properties	O
of	O
human	B-Organism
oral	B-Cell
squamous	I-Cell
cell	I-Cell
carcinoma	I-Cell
cells	I-Cell
.	O

Little	O
is	O
known	O
about	O
the	O
specificity	O
of	O
angiogenic	O
properties	O
of	O
oral	B-Cell
cancer	I-Cell
cells	I-Cell
and	O
the	O
possible	O
mechanisms	O
.	O

Stimulatory	O
effects	O
on	O
proliferation	O
and	O
migration	O
of	O
human	B-Cell
umbilical	I-Cell
vein	I-Cell
endothelial	I-Cell
cells	I-Cell
(	O
HUVEC	B-Cell
)	O
characterized	O
the	O
angiogenic	O
properties	O
of	O
oral	B-Cell
cancer	I-Cell
cells	I-Cell
but	O
not	O
normal	B-Cell
oral	I-Cell
keratinocytes	I-Cell
(	O
NOK	B-Cell
)	O
.	O

ELISA	O
found	O
the	O
presence	O
of	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factors	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
both	O
in	O
the	O
tested	O
oral	B-Cell
cancer	I-Cell
cells	I-Cell
and	O
NOK	B-Cell
.	O

Attenuation	O
of	O
the	O
proangiogenic	O
effects	O
by	O
neutralizing	O
VEGF	B-Gene_or_gene_product
antibodies	I-Gene_or_gene_product
suggests	O
VEGF	B-Gene_or_gene_product
play	O
a	O
key	O
role	O
in	O
the	O
acquisition	O
of	O
the	O
angiogenic	O
phenotype	O
in	O
oral	B-Cell
cancer	I-Cell
cells	I-Cell
.	O

Western	O
blotting	O
of	O
p53	B-Gene_or_gene_product
and	O
murine	B-Gene_or_gene_product
double	I-Gene_or_gene_product
mutant	I-Gene_or_gene_product
2	I-Gene_or_gene_product
(	O
Mdm2	B-Gene_or_gene_product
)	O
together	O
with	O
p53	B-Gene_or_gene_product
DNA	B-Cellular_component
sequencing	O
analysis	O
indicate	O
that	O
p53	B-Gene_or_gene_product
function	O
loss	O
by	O
mutation	O
or	O
overexpression	O
of	O
Mdm2	B-Gene_or_gene_product
occurred	O
in	O
all	O
tested	O
oral	B-Cell
cancer	I-Cell
cells	I-Cell
regardless	O
of	O
their	O
etiology	O
.	O

In	O
summary	O
,	O
the	O
angiogenic	O
property	O
of	O
oral	B-Cell
cancer	I-Cell
cells	I-Cell
is	O
mediated	O
by	O
many	O
factors	O
in	O
addition	O
to	O
VEGF	B-Gene_or_gene_product
and	O
the	O
functional	O
status	O
of	O
p53	B-Gene_or_gene_product
.	O

Enforced	O
expression	O
of	O
tissue	B-Gene_or_gene_product
inhibitor	I-Gene_or_gene_product
of	I-Gene_or_gene_product
matrix	I-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
affects	O
functional	O
capillary	B-Tissue
morphogenesis	O
and	O
inhibits	O
tumor	B-Cancer
growth	O
in	O
a	O
murine	B-Organism
tumor	B-Cancer
model	O
.	O

Homeostasis	O
of	O
the	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
is	O
a	O
delicate	O
balance	O
between	O
degradation	O
and	O
remodeling	O
,	O
the	O
balance	O
being	O
maintained	O
by	O
the	O
interaction	O
of	O
activated	O
matrix	B-Gene_or_gene_product
metalloproteinases	I-Gene_or_gene_product
(	O
MMPs	B-Gene_or_gene_product
)	O
and	O
specific	O
tissue	B-Gene_or_gene_product
inhibitors	I-Gene_or_gene_product
of	I-Gene_or_gene_product
matrix	I-Gene_or_gene_product
metalloproteinases	I-Gene_or_gene_product
(	O
TIMPs	B-Gene_or_gene_product
)	O
.	O

Up	O
-	O
regulation	O
of	O
MMP	B-Gene_or_gene_product
activity	O
,	O
favoring	O
proteolytic	O
degradation	O
of	O
the	O
basement	B-Cellular_component
membrane	I-Cellular_component
and	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
,	O
has	O
been	O
linked	O
to	O
tumor	B-Cancer
growth	O
and	O
metastasis	O
,	O
as	O
well	O
as	O
tumor	B-Cancer
-	O
associated	O
angiogenesis	O
,	O
whereas	O
inhibition	O
of	O
MMP	B-Gene_or_gene_product
activity	O
appears	O
to	O
restrict	O
these	O
processes	O
.	O

We	O
have	O
used	O
retroviral	O
-	O
mediated	O
gene	O
delivery	O
to	O
effect	O
sustained	O
autocrine	O
expression	O
of	O
TIMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
in	O
murine	B-Organism
neuroblastoma	B-Cell
and	O
melanoma	B-Cell
tumor	I-Cell
cells	I-Cell
in	O
order	O
to	O
further	O
examine	O
the	O
ability	O
of	O
TIMPs	B-Gene_or_gene_product
to	O
inhibit	O
angiogenesis	O
in	O
vivo	O
.	O

Growth	O
of	O
both	O
histologic	O
types	O
of	O
gene	O
-	O
modified	O
tumor	B-Cell
cells	I-Cell
in	O
severe	O
combined	O
immunodeficiency	O
(	O
SCID	O
)	O
mice	B-Organism
was	O
significantly	O
restricted	O
when	O
compared	O
with	O
controls	O
.	O

Grossly	O
,	O
these	O
tumors	B-Cancer
were	O
small	O
and	O
had	O
few	O
feeding	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
.	O

Histologic	O
evaluation	O
revealed	O
that	O
although	O
tumors	B-Cancer
overexpressing	O
TIMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
had	O
an	O
increased	O
number	O
of	O
CD31	B-Gene_or_gene_product
(	O
+	O
)	O
endothelial	O
cells	O
,	O
these	O
endothelial	B-Cell
cells	I-Cell
had	O
not	O
formed	O
functional	O
tubules	B-Tissue
,	O
as	O
evidenced	O
by	O
decreased	O
vessel	B-Multi-tissue_structure
continuity	O
and	O
minimal	O
pericyte	B-Cell
recruitment	O
.	O

This	O
effect	O
appears	O
to	O
be	O
mediated	O
,	O
in	O
part	O
,	O
by	O
decreased	O
expression	O
of	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
(	I-Gene_or_gene_product
VE	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
by	O
endothelial	B-Cell
cells	I-Cell
in	O
the	O
presence	O
of	O
TIMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
as	O
seen	O
both	O
in	O
an	O
in	O
vitro	O
assay	O
and	O
in	O
TIMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
-	O
overexpressing	O
tumors	B-Cancer
.	O

Taken	O
together	O
,	O
these	O
results	O
demonstrate	O
that	O
overexpression	O
of	O
TIMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
can	O
inhibit	O
angiogenesis	O
and	O
associated	O
tumor	B-Cancer
growth	O
,	O
and	O
that	O
the	O
antiangiogenic	O
effects	O
of	O
TIMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
appear	O
to	O
be	O
mediated	O
through	O
the	O
inhibition	O
of	O
functional	O
capillary	B-Tissue
morphogenesis	O
.	O

Brn	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3a	I-Gene_or_gene_product
,	O
a	O
neuronal	B-Cell
transcription	O
factor	O
of	O
the	O
POU	O
gene	O
family	O
:	O
indications	O
for	O
its	O
involvement	O
in	O
cancer	B-Cancer
and	O
angiogenesis	O
.	O

Brn	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3a	I-Gene_or_gene_product
,	O
a	O
member	O
of	O
the	O
POU	O
gene	O
family	O
(	O
so	O
-	O
called	O
because	O
of	O
the	O
similarity	O
with	O
the	O
group	O
of	O
transcription	O
factors	O
Pit	B-Gene_or_gene_product
,	O
Oct	B-Gene_or_gene_product
,	O
and	O
Unc	B-Gene_or_gene_product
)	O
,	O
was	O
found	O
in	O
neuronal	B-Cell
cells	I-Cell
engaged	O
in	O
the	O
transcription	O
activity	O
of	O
the	O
p1	O
and	O
p2	O
promoters	O
of	O
the	O
most	O
powerful	O
antiapoptotic	O
gene	O
,	O
namely	O
,	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
.	O

The	O
alternative	O
splicing	O
of	O
Brn	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3a	I-Gene_or_gene_product
mRNA	O
produces	O
two	O
molecular	O
forms	O
:	O
a	O
longer	O
,	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
transactivating	O
form	O
,	O
and	O
a	O
shorter	O
inactive	O
form	O
,	O
lacking	O
84	O
AA	O
in	O
the	O
aminoterminus	O
.	O

In	O
neuronal	B-Cell
cells	I-Cell
,	O
following	O
Brn	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3a	I-Gene_or_gene_product
gene	O
transfection	O
and	O
superexpression	O
,	O
an	O
increase	O
of	O
30	O
fold	O
of	O
the	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
protein	O
occurs	O
,	O
leading	O
to	O
apoptosis	O
protection	O
.	O

However	O
,	O
recent	O
works	O
demonstrate	O
that	O
Brn	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3a	I-Gene_or_gene_product
expression	O
is	O
not	O
restricted	O
to	O
neuronal	B-Cell
cells	I-Cell
,	O
as	O
its	O
activity	O
was	O
detected	O
also	O
in	O
cancer	B-Cell
cells	I-Cell
of	O
non	O
-	O
neuronal	B-Cell
nature	O
.	O

Looking	O
for	O
mechanisms	O
linking	O
Brn	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3a	I-Gene_or_gene_product
to	O
carcinogenesis	O
,	O
we	O
discuss	O
the	O
role	O
of	O
this	O
transcription	O
factor	O
in	O
influencing	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
/	O
p53	B-Gene_or_gene_product
antagonism	O
and	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
/	O
VEGF	B-Gene_or_gene_product
induction	O
of	O
tumor	B-Cancer
angiogenesis	O
,	O
concluding	O
this	O
review	O
with	O
a	O
proposal	O
for	O
the	O
oncogenic	O
nature	O
of	O
Brn	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3a	I-Gene_or_gene_product
.	O

Roles	O
of	O
cell	B-Cell
adhesion	O
molecules	O
in	O
tumor	B-Cancer
angiogenesis	O
induced	O
by	O
cotransplantation	O
of	O
cancer	B-Cell
and	O
endothelial	B-Cell
cells	I-Cell
to	O
nude	B-Organism
rats	I-Organism
.	O

Roles	O
of	O
cell	B-Cell
adhesion	O
molecules	O
mediating	O
the	O
interaction	O
of	O
cancer	B-Cell
and	O
endothelial	B-Cell
cells	I-Cell
in	O
tumor	B-Cancer
angiogenesis	O
were	O
investigated	O
using	O
new	O
in	O
vitro	O
and	O
in	O
vivo	O
model	O
systems	O
with	O
a	O
cultured	O
murine	B-Organism
endothelial	B-Cell
cell	I-Cell
line	I-Cell
(	O
F	B-Cell
-	I-Cell
2	I-Cell
)	O
and	O
human	B-Organism
cultured	O
epidermoid	B-Cell
cancer	I-Cell
cells	I-Cell
(	O
A431	B-Cell
)	O
.	O

The	O
A431	B-Cell
cells	I-Cell
exhibited	O
typical	O
in	O
vitro	O
cell	B-Cell
adhesion	O
to	O
the	O
endothelial	B-Cell
F	I-Cell
-	I-Cell
2	I-Cell
cells	I-Cell
.	O

The	O
initial	O
step	O
of	O
adhesion	O
was	O
mediated	O
by	O
sialyl	B-Simple_chemical
Lewis	I-Simple_chemical
(	I-Simple_chemical
x	I-Simple_chemical
)	I-Simple_chemical
(	O
Le	B-Simple_chemical
(	I-Simple_chemical
x	I-Simple_chemical
)	I-Simple_chemical
)	O
and	O
sialyl	B-Simple_chemical
Le	I-Simple_chemical
(	I-Simple_chemical
a	I-Simple_chemical
)	I-Simple_chemical
,	O
the	O
carbohydrate	O
determinants	O
expressed	O
on	O
the	O
cancer	B-Cell
cells	I-Cell
,	O
and	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
selectin	I-Gene_or_gene_product
expressed	O
constitutively	O
on	O
F	B-Cell
-	I-Cell
2	I-Cell
cells	I-Cell
.	O

Prolonged	O
culture	O
led	O
to	O
the	O
implantation	O
of	O
cancer	B-Cell
cells	I-Cell
into	O
the	O
monolayer	B-Cell
of	O
the	O
F	B-Cell
-	I-Cell
2	I-Cell
cells	I-Cell
,	O
which	O
was	O
mediated	O
mainly	O
by	O
alpha	B-Gene_or_gene_product
(	I-Gene_or_gene_product
3	I-Gene_or_gene_product
)	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
(	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	I-Gene_or_gene_product
integrin	I-Gene_or_gene_product
.	O

F	B-Cell
-	I-Cell
2	I-Cell
cells	I-Cell
cultured	O
on	O
Matrigel	O
showed	O
evident	O
tube	B-Tissue
formation	O
,	O
and	O
coculture	O
of	O
F	B-Cell
-	I-Cell
2	I-Cell
cells	I-Cell
with	O
A431	B-Cell
cells	I-Cell
led	O
to	O
the	O
formation	O
of	O
A431	B-Cell
cell	I-Cell
nests	O
constantly	O
surrounded	O
by	O
tube	B-Tissue
-	O
like	O
networks	O
consisting	O
of	O
F	B-Cell
-	I-Cell
2	I-Cell
cells	I-Cell
.	O

This	O
in	O
vitro	O
morphogenesis	O
was	O
inhibited	O
by	O
the	O
addition	O
of	O
anti	O
-	O
sialyl	B-Simple_chemical
Le	I-Simple_chemical
(	I-Simple_chemical
x	I-Simple_chemical
)	I-Simple_chemical
/	O
Le	B-Simple_chemical
(	I-Simple_chemical
a	I-Simple_chemical
)	I-Simple_chemical
or	O
anti	O
-	O
beta	B-Gene_or_gene_product
(	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	I-Gene_or_gene_product
integrin	I-Gene_or_gene_product
antibodies	O
,	O
which	O
led	O
to	O
the	O
formation	O
of	O
cancer	B-Cell
cell	I-Cell
aggregates	I-Cell
that	O
were	O
independent	O
from	O
the	O
F	B-Cell
-	I-Cell
2	I-Cell
cell	I-Cell
networks	O
.	O

This	O
in	O
vitro	O
morphological	O
appearance	O
was	O
exactly	O
reproduced	O
in	O
the	O
in	O
vivo	O
tumors	B-Cancer
,	O
which	O
were	O
formed	O
when	O
the	O
mixture	O
of	O
A431	B-Cell
and	O
F	B-Cell
-	I-Cell
2	I-Cell
cells	I-Cell
at	O
the	O
ratio	O
of	O
10	O
:	O
1	O
were	O
cotransplanted	O
s	O
.	O
c	O
.	O
into	O
the	O
back	B-Organism_subdivision
of	O
nude	B-Organism
rats	I-Organism
.	O

The	O
tumors	B-Cancer
of	O
A431	B-Cell
supplemented	O
with	O
F	B-Cell
-	I-Cell
2	I-Cell
cells	I-Cell
were	O
profoundly	O
vascularized	O
throughout	O
by	O
the	O
tubular	B-Tissue
structures	I-Tissue
formed	O
by	O
F	B-Cell
-	I-Cell
2	I-Cell
cells	I-Cell
,	O
the	O
lumen	B-Immaterial_anatomical_entity
of	O
which	O
contained	O
the	O
host	O
rat	B-Organism
blood	B-Cell
cells	I-Cell
.	O

The	O
tumor	B-Cancer
mass	O
thus	O
formed	O
was	O
an	O
average	O
5	O
.	O
8	O
-	O
fold	O
as	O
large	O
as	O
control	O
A431	B-Cancer
tumors	I-Cancer
that	O
were	O
grown	O
without	O
F	B-Cell
-	I-Cell
2	I-Cell
cells	I-Cell
.	O

The	O
co	O
-	O
injection	O
of	O
anti	O
-	O
Le	B-Simple_chemical
(	I-Simple_chemical
x	I-Simple_chemical
)	I-Simple_chemical
/	O
Le	B-Simple_chemical
(	I-Simple_chemical
a	I-Simple_chemical
)	I-Simple_chemical
or	O
anti	O
-	O
beta	B-Gene_or_gene_product
(	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	I-Gene_or_gene_product
integrin	I-Gene_or_gene_product
antibodies	O
produced	O
a	O
marked	O
reduction	O
in	O
the	O
size	O
of	O
A431	B-Cancer
tumors	I-Cancer
,	O
which	O
were	O
not	O
vascularized	O
and	O
accompanied	O
an	O
independent	O
tiny	O
remnant	O
clump	O
of	O
F	B-Cell
-	I-Cell
2	I-Cell
cells	I-Cell
.	O

The	O
size	O
of	O
these	O
A431	B-Cancer
tumors	I-Cancer
did	O
not	O
differ	O
significantly	O
from	O
those	O
of	O
control	O
A431	B-Cancer
tumors	I-Cancer
raised	O
without	O
F	B-Cell
-	I-Cell
2	I-Cell
cells	I-Cell
.	O

These	O
results	O
indicate	O
that	O
the	O
interaction	O
of	O
tumor	B-Cell
cells	I-Cell
and	O
endothelial	B-Cell
cells	I-Cell
in	O
orderly	O
tumor	B-Cancer
angiomorphogenesis	O
is	O
highly	O
dependent	O
on	O
the	O
action	O
of	O
cell	B-Cell
adhesion	O
molecules	O
mediating	O
the	O
adhesion	O
of	O
cancer	B-Cell
cells	I-Cell
to	O
endothelial	B-Cell
cells	I-Cell
,	O
inhibition	O
of	O
which	O
remarkably	O
retards	O
tumor	B-Cancer
growth	O
and	O
angiogenesis	O
.	O

Identification	O
of	O
Dss1	B-Gene_or_gene_product
as	O
a	O
12	B-Simple_chemical
-	I-Simple_chemical
O	I-Simple_chemical
-	I-Simple_chemical
tetradecanoylphorbol	I-Simple_chemical
-	I-Simple_chemical
13	I-Simple_chemical
-	I-Simple_chemical
acetate	I-Simple_chemical
-	O
responsive	O
gene	O
expressed	O
in	O
keratinocyte	B-Cell
progenitor	I-Cell
cells	I-Cell
,	O
with	O
possible	O
involvement	O
in	O
early	O
skin	B-Organ
tumorigenesis	O
.	O

This	O
study	O
identifies	O
genes	O
expressed	O
early	O
in	O
12	B-Simple_chemical
-	I-Simple_chemical
O	I-Simple_chemical
-	I-Simple_chemical
tetradecanoylphorbol	I-Simple_chemical
-	I-Simple_chemical
13	I-Simple_chemical
-	I-Simple_chemical
acetate	I-Simple_chemical
(	O
TPA	B-Simple_chemical
)	O
-	O
induced	O
skin	B-Organ
carcinogenesis	O
in	O
genetically	O
initiated	O
Tg	O
.	O
AC	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Ha	I-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
transgenic	O
mice	O
.	O

Keratinocyte	B-Cell
progenitor	I-Cell
cells	I-Cell
from	O
TPA	B-Simple_chemical
-	O
treated	O
Tg	B-Organism
.	I-Organism
AC	I-Organism
mice	I-Organism
were	O
isolated	O
with	O
fluorescence	O
-	O
activated	O
cell	B-Cell
sorting	O
and	O
expression	O
was	O
analyzed	O
using	O
cDNA	O
microarray	O
technology	O
.	O

Eleven	O
genes	O
were	O
identified	O
whose	O
expression	O
changed	O
significantly	O
in	O
response	O
to	O
carcinogen	O
treatment	O
.	O

Deleted	B-Gene_or_gene_product
in	I-Gene_or_gene_product
split	I-Gene_or_gene_product
hand	I-Gene_or_gene_product
/	I-Gene_or_gene_product
split	I-Gene_or_gene_product
foot	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
Dss1	B-Gene_or_gene_product
)	O
is	O
a	O
gene	O
associated	O
with	O
a	O
heterogeneous	O
limb	B-Organism_subdivision
developmental	O
disorder	O
called	O
split	O
hand	B-Organism_subdivision
/	O
split	O
foot	B-Organism_subdivision
malformation	O
.	O

cDNA	O
microarray	O
expression	O
analysis	O
showed	O
that	O
the	O
mouse	B-Organism
homologue	O
of	O
Dss1	B-Gene_or_gene_product
is	O
induced	O
by	O
TPA	B-Simple_chemical
.	O

Dss1	B-Gene_or_gene_product
overexpression	O
was	O
detected	O
by	O
Northern	O
blot	O
analysis	O
in	O
early	O
TPA	B-Simple_chemical
-	O
treated	O
hyperplastic	O
skins	B-Organ
and	O
in	O
JB6	B-Cell
Cl	I-Cell
41	I-Cell
-	I-Cell
5a	I-Cell
epidermal	I-Cell
cells	I-Cell
.	O

Interestingly	O
,	O
Dss1	B-Gene_or_gene_product
expression	O
was	O
also	O
shown	O
to	O
be	O
elevated	O
in	O
skin	B-Organ
papillomas	I-Organ
relative	O
to	O
normal	O
skins	B-Organ
,	O
and	O
further	O
increased	O
in	O
squamous	B-Cell
cell	I-Cell
malignancies	I-Cell
.	O

Functional	O
studies	O
by	O
ectopically	O
constitutive	O
expression	O
of	O
Dss1	B-Gene_or_gene_product
in	O
JB6	B-Cell
Cl	I-Cell
41	I-Cell
-	I-Cell
5a	I-Cell
preneoplastic	I-Cell
cells	I-Cell
strongly	O
increased	O
focus	O
formation	O
and	O
proliferation	O
of	O
these	O
cells	B-Cell
and	O
enhanced	O
efficiency	O
of	O
neoplastic	O
transformation	O
of	O
the	O
cells	B-Cell
in	O
soft	O
agar	O
.	O

These	O
results	O
strongly	O
suggest	O
that	O
Dss1	B-Gene_or_gene_product
is	O
a	O
TPA	B-Simple_chemical
-	O
inducible	O
gene	O
that	O
may	O
play	O
an	O
important	O
role	O
in	O
the	O
early	O
stages	O
of	O
skin	B-Organ
carcinogenesis	O
.	O

Downregulation	O
of	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
FLIP	I-Gene_or_gene_product
sensitizes	O
DU145	B-Cell
prostate	I-Cell
cancer	I-Cell
cells	I-Cell
to	O
Fas	B-Gene_or_gene_product
-	O
mediated	O
apoptosis	O
.	O

Although	O
DU145	B-Cell
prostate	I-Cell
cancer	I-Cell
cells	I-Cell
are	O
resistant	O
to	O
exogenously	O
applied	O
Fas	B-Gene_or_gene_product
agonist	O
CH	O
-	O
11	O
(	O
anti	O
-	O
Fas	B-Gene_or_gene_product
monoclonal	O
antibody	O
)	O
,	O
Fas	B-Gene_or_gene_product
-	O
resistance	O
can	O
be	O
overcome	O
using	O
a	O
FasL	B-Gene_or_gene_product
expressing	O
adenovirus	B-Organism
(	O
AdGFPFasL	B-Organism
(	I-Organism
TET	I-Organism
)	I-Organism
)	O
[	O
Hyer	O
et	O
al	O
.	O
,	O
Molecular	O
Therapy	O
,	O
2000	O
;	O
2	O
:	O
348	O
-	O
58	O
(	O
ref	O
.	O
12	O
)	O
]	O
.	O

The	O
purpose	O
of	O
this	O
study	O
was	O
to	O
try	O
to	O
understand	O
why	O
DU145	B-Cell
cells	I-Cell
are	O
resistant	O
to	O
CH	O
-	O
11	O
and	O
determine	O
the	O
signaling	O
pathway	O
utilized	O
by	O
AdGFPFasL	B-Organism
(	I-Organism
TET	I-Organism
)	I-Organism
to	O
induce	O
apoptosis	O
in	O
these	O
Fas	B-Gene_or_gene_product
-	O
resistant	O
cells	B-Cell
.	O

Using	O
immunoblot	O
analysis	O
,	O
we	O
show	O
that	O
AdGFPFasL	B-Organism
(	I-Organism
TET	I-Organism
)	I-Organism
is	O
capable	O
of	O
initiating	O
the	O
classic	O
Fas	B-Gene_or_gene_product
-	O
mediated	O
apoptotic	O
pathway	O
in	O
DU145	B-Cell
cells	I-Cell
,	O
which	O
includes	O
activation	O
of	O
caspases	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
,	O
-	B-Gene_or_gene_product
3	I-Gene_or_gene_product
,	O
-	B-Gene_or_gene_product
7	I-Gene_or_gene_product
,	O
and	O
-	B-Gene_or_gene_product
9	I-Gene_or_gene_product
,	O
BID	B-Gene_or_gene_product
cleavage	O
,	O
cytochrome	B-Gene_or_gene_product
c	I-Gene_or_gene_product
release	O
from	O
mitochondria	B-Cellular_component
,	O
and	O
PARP	B-Gene_or_gene_product
cleavage	O
.	O

In	O
contrast	O
,	O
CH	O
-	O
11	O
binds	O
to	O
Fas	B-Gene_or_gene_product
,	O
but	O
is	O
unable	O
to	O
transmit	O
the	O
death	O
signal	O
beyond	O
the	O
plasma	B-Cellular_component
membrane	I-Cellular_component
suggesting	O
a	O
block	O
at	O
the	O
DISC	B-Gene_or_gene_product
(	O
death	B-Gene_or_gene_product
inducing	I-Gene_or_gene_product
signaling	I-Gene_or_gene_product
complex	I-Gene_or_gene_product
)	O
.	O

The	O
anti	O
-	O
apoptotic	O
protein	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
FLIP	I-Gene_or_gene_product
(	O
cellular	B-Gene_or_gene_product
Flice	I-Gene_or_gene_product
-	I-Gene_or_gene_product
like	I-Gene_or_gene_product
inhibitory	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
)	O
,	O
which	O
has	O
been	O
shown	O
to	O
inhibit	O
Fas	B-Gene_or_gene_product
-	O
mediated	O
apoptosis	O
at	O
the	O
DISC	B-Gene_or_gene_product
,	O
was	O
down	O
-	O
regulated	O
following	O
AdGFPFasL	B-Organism
(	I-Organism
TET	I-Organism
)	I-Organism
treatment	O
prompting	O
us	O
to	O
investigate	O
its	O
role	O
in	O
inhibiting	O
CH	O
-	O
11	O
-	O
induced	O
cell	B-Cell
death	O
.	O

Using	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
FLIP	I-Gene_or_gene_product
anti	O
-	O
sense	O
oligonucleotides	O
to	O
down	O
-	O
regulate	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
FLIP	I-Gene_or_gene_product
we	O
sensitized	O
DU145	B-Cell
cells	I-Cell
to	O
CH	O
-	O
11	O
-	O
induced	O
apoptosis	O
.	O

These	O
data	O
suggest	O
that	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
FLIP	I-Gene_or_gene_product
may	O
play	O
a	O
critical	O
role	O
in	O
regulating	O
Fas	B-Gene_or_gene_product
-	O
mediated	O
apoptosis	O
in	O
prostate	B-Cell
cancer	I-Cell
cells	I-Cell
and	O
that	O
modulation	O
of	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
FLIP	I-Gene_or_gene_product
may	O
enhance	O
Fas	B-Gene_or_gene_product
signaling	O
based	O
therapies	O
.	O

Topoisomerase	B-Gene_or_gene_product
I	I-Gene_or_gene_product
protein	O
expression	O
in	O
primary	B-Cancer
colorectal	I-Cancer
cancer	I-Cancer
and	O
lymph	B-Cancer
node	I-Cancer
metastases	I-Cancer
.	O

Topoisomerase	B-Gene_or_gene_product
I	I-Gene_or_gene_product
(	O
topo	B-Gene_or_gene_product
I	I-Gene_or_gene_product
)	O
is	O
an	O
important	O
target	O
for	O
the	O
treatment	O
of	O
malignant	B-Cancer
disease	I-Cancer
,	O
especially	O
colorectal	B-Cancer
cancer	I-Cancer
.	O

Because	O
there	O
is	O
little	O
information	O
on	O
the	O
expression	O
of	O
topo	B-Gene_or_gene_product
I	I-Gene_or_gene_product
in	O
colorectal	B-Cancer
tumors	I-Cancer
,	O
this	O
study	O
evaluated	O
and	O
characterized	O
topo	B-Gene_or_gene_product
I	I-Gene_or_gene_product
protein	O
expression	O
in	O
primary	O
colorectal	B-Cancer
cancer	I-Cancer
and	O
lymph	B-Cancer
node	I-Cancer
metastases	I-Cancer
and	O
studied	O
the	O
association	O
between	O
topo	B-Gene_or_gene_product
I	I-Gene_or_gene_product
protein	O
expression	O
and	O
clinicopathologic	O
data	O
,	O
p53	B-Gene_or_gene_product
status	O
,	O
and	O
proliferating	B-Gene_or_gene_product
cell	I-Gene_or_gene_product
nuclear	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
(	O
PCNA	B-Gene_or_gene_product
)	O
status	O
.	O

Immunohistochemistry	O
assay	O
was	O
performed	O
for	O
topo	B-Gene_or_gene_product
I	I-Gene_or_gene_product
protein	O
expression	O
in	O
249	O
primary	O
human	B-Organism
colorectal	B-Cancer
cancer	I-Cancer
and	O
42	O
paired	O
lymph	B-Cancer
node	I-Cancer
metastasis	I-Cancer
samples	I-Cancer
.	O

Topo	B-Gene_or_gene_product
I	I-Gene_or_gene_product
expression	O
was	O
described	O
as	O
the	O
percentage	O
of	O
cells	B-Cell
staining	O
positive	O
for	O
topo	B-Gene_or_gene_product
I	I-Gene_or_gene_product
,	O
along	O
with	O
the	O
intensity	O
and	O
localization	O
of	O
the	O
staining	O
.	O

Clinicopathologic	O
data	O
(	O
sex	O
,	O
age	O
,	O
Dukes	O
'	O
stage	O
,	O
differentiation	O
grade	O
,	O
survival	O
status	O
)	O
,	O
p53	B-Gene_or_gene_product
status	O
,	O
and	O
PCNA	B-Gene_or_gene_product
status	O
were	O
statistically	O
analyzed	O
for	O
association	O
with	O
topo	B-Gene_or_gene_product
I	I-Gene_or_gene_product
protein	O
expression	O
.	O

Topo	B-Gene_or_gene_product
I	I-Gene_or_gene_product
expression	O
in	O
paired	O
primary	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastases	O
were	O
studied	O
for	O
concordance	O
.	O

Topo	B-Gene_or_gene_product
I	I-Gene_or_gene_product
protein	O
expression	O
was	O
detected	O
in	O
127	O
(	O
51	O
%	O
)	O
samples	B-Cancer
,	O
including	O
24	O
.	O
4	O
%	O
with	O
>	O
50	O
%	O
positive	O
tumor	B-Cell
cells	I-Cell
.	O

The	O
majority	O
had	O
nuclear	B-Cellular_component
(	O
70	O
.	O
1	O
%	O
)	O
or	O
nuclear	B-Cellular_component
and	O
cytoplasmic	B-Organism_substance
staining	O
(	O
17	O
.	O
3	O
%	O
)	O
.	O

A	O
higher	O
percentage	O
of	O
cells	B-Cell
expressing	O
topo	B-Gene_or_gene_product
I	I-Gene_or_gene_product
in	O
primary	B-Cancer
colorectal	I-Cancer
cancer	I-Cancer
was	O
significantly	O
associated	O
with	O
advanced	O
age	O
(	O
P	O
=	O
.	O
040	O
)	O
.	O

Patients	B-Organism
with	O
rectal	B-Cancer
cancer	I-Cancer
had	O
greater	O
topo	B-Gene_or_gene_product
I	I-Gene_or_gene_product
expression	O
than	O
those	O
with	O
colon	B-Cancer
tumors	I-Cancer
(	O
P	O
=	O
.	O
029	O
)	O
.	O

No	O
significant	O
correlation	O
was	O
found	O
between	O
topo	B-Gene_or_gene_product
I	I-Gene_or_gene_product
protein	O
expression	O
and	O
sex	O
,	O
Dukes	O
'	O
stage	O
,	O
differentiation	O
grade	O
,	O
survival	O
status	O
,	O
p53	B-Gene_or_gene_product
status	O
,	O
and	O
PCNA	B-Gene_or_gene_product
status	O
.	O

Concordance	O
in	O
topo	B-Gene_or_gene_product
I	I-Gene_or_gene_product
staining	O
between	O
primary	B-Cancer
and	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastases	O
was	O
observed	O
in	O
33	O
of	O
42	O
cases	O
(	O
P	O
=	O
.	O
029	O
)	O
.	O

This	O
suggests	O
that	O
the	O
activity	O
of	O
topo	B-Gene_or_gene_product
I	I-Gene_or_gene_product
inhibitors	O
will	O
not	O
differ	O
across	O
various	O
tumor	B-Cancer
stages	O
,	O
pathology	O
,	O
and	O
patient	B-Organism
gender	O
.	O

p53	B-Gene_or_gene_product
and	O
PCNA	B-Gene_or_gene_product
status	O
do	O
not	O
appear	O
to	O
influence	O
topo	B-Gene_or_gene_product
I	I-Gene_or_gene_product
expression	O
,	O
and	O
topo	B-Gene_or_gene_product
I	I-Gene_or_gene_product
has	O
no	O
apparent	O
association	O
with	O
the	O
acquisition	O
of	O
a	O
metastatic	O
phenotype	O
.	O

Topo	B-Gene_or_gene_product
I	I-Gene_or_gene_product
expression	O
now	O
needs	O
to	O
be	O
evaluated	O
in	O
patients	B-Organism
undergoing	O
topo	B-Gene_or_gene_product
I	I-Gene_or_gene_product
-	O
inhibitor	O
therapy	O
,	O
to	O
better	O
define	O
the	O
role	O
of	O
this	O
protein	O
as	O
a	O
predictive	O
marker	O
.	O

Regulation	O
of	O
transforming	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
signaling	O
and	O
vascular	B-Multi-tissue_structure
diseases	O
.	O

PURPOSE	O
:	O
Members	O
of	O
the	O
transforming	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	I-Gene_or_gene_product
TGF	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
superfamily	O
play	O
critical	O
roles	O
in	O
regulation	O
of	O
various	O
cellular	B-Cell
functions	O
.	O

Dysregulation	O
of	O
the	O
signaling	O
mechanisms	O
of	O
the	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
superfamily	O
proteins	O
is	O
associated	O
with	O
clinical	O
diseases	O
such	O
as	O
cancer	B-Cancer
,	O
fibrotic	O
diseases	O
,	O
and	O
vascular	B-Multi-tissue_structure
disorders	O
.	O

Therefore	O
,	O
understanding	O
these	O
signaling	O
mechanisms	O
may	O
provide	O
us	O
with	O
novel	O
ways	O
to	O
develop	O
strategies	O
for	O
treating	O
clinical	O
diseases	O
induced	O
by	O
these	O
cytokines	O
.	O

METHODS	O
:	O
This	O
review	O
discusses	O
our	O
current	O
understanding	O
of	O
the	O
mechanisms	O
of	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
signaling	O
,	O
focusing	O
on	O
the	O
roles	O
of	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
in	O
regulation	O
of	O
vascular	B-Cell
wall	I-Cell
cells	I-Cell
and	O
on	O
the	O
regulation	O
of	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
superfamily	O
signals	O
by	O
inhibitory	O
Smads	B-Gene_or_gene_product
.	O

[	O
Expression	O
and	O
mutation	O
of	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kit	I-Gene_or_gene_product
gene	O
in	O
gastrointestinal	B-Cancer
stromal	I-Cancer
tumor	I-Cancer
]	O
.	O

OBJECTIVE	O
:	O
To	O
investigate	O
the	O
effect	O
of	O
the	O
expression	O
and	O
mutation	O
of	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kit	I-Gene_or_gene_product
gene	O
and	O
its	O
relationship	O
with	O
clinical	O
pathology	O
and	O
prognosis	O
of	O
gastrointestinal	B-Cancer
stromal	I-Cancer
tumor	I-Cancer
(	O
GIST	B-Cancer
)	O
.	O

METHODS	O
:	O
Immunohistochemical	O
and	O
PCR	O
-	O
SSCP	O
techniques	O
were	O
used	O
to	O
detect	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kit	I-Gene_or_gene_product
protein	O
expression	O
and	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kit	I-Gene_or_gene_product
gene	O
exon	O
11	O
mutation	O
in	O
82	O
patients	B-Organism
with	O
GIST	B-Cancer
.	O

RESULTS	O
:	O
The	O
positive	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kit	I-Gene_or_gene_product
protein	O
expression	O
and	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kit	I-Gene_or_gene_product
gene	O
mutation	O
rates	O
were	O
97	O
.	O
6	O
%	O
(	O
80	O
/	O
82	O
)	O
and	O
41	O
.	O
5	O
%	O
(	O
34	O
/	O
82	O
)	O
.	O

Correlating	O
the	O
results	O
of	O
these	O
two	O
methods	O
and	O
clinicopathological	O
factors	O
,	O
the	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kit	I-Gene_or_gene_product
expression	O
and	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kit	I-Gene_or_gene_product
gene	O
mutation	O
rates	O
were	O
95	O
.	O
0	O
%	O
(	O
19	O
/	O
20	O
)	O
and	O
0	O
in	O
benign	B-Cancer
GIST	I-Cancer
,	O
and	O
were	O
98	O
.	O
4	O
%	O
(	O
61	O
/	O
62	O
)	O
,	O
54	O
.	O
8	O
%	O
(	O
34	O
/	O
62	O
)	O
in	O
malignant	B-Cancer
GIST	I-Cancer
.	O

Mutation	O
positive	O
GIST	B-Cancer
showed	O
higher	O
frequency	O
of	O
adjacent	O
tissue	B-Tissue
invasion	O
,	O
metastasis	O
and	O
recurrence	O
as	O
compared	O
with	O
mutation	O
negative	O
ones	O
.	O

CONCLUSION	O
:	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kit	I-Gene_or_gene_product
protein	O
is	O
an	O
important	O
diagnostic	O
marker	O
of	O
gastrointestinal	B-Cancer
stromal	I-Cancer
tumor	I-Cancer
.	O

c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kit	I-Gene_or_gene_product
gene	O
mutation	O
may	O
play	O
a	O
significant	O
role	O
in	O
the	O
pathogenesis	O
of	O
GIST	B-Cancer
and	O
also	O
may	O
be	O
a	O
prognostic	O
marker	O
.	O

Inhibition	O
of	O
glucose	B-Simple_chemical
metabolism	O
sensitizes	O
tumor	B-Cell
cells	I-Cell
to	O
death	O
receptor	O
-	O
triggered	O
apoptosis	O
through	O
enhancement	O
of	O
death	B-Gene_or_gene_product
-	I-Gene_or_gene_product
inducing	I-Gene_or_gene_product
signaling	I-Gene_or_gene_product
complex	I-Gene_or_gene_product
formation	O
and	O
apical	O
procaspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
processing	O
.	O

Tumors	B-Cancer
display	O
a	O
high	O
rate	O
of	O
glucose	B-Simple_chemical
uptake	O
and	O
glycolysis	O
.	O

We	O
investigated	O
how	O
inhibition	O
of	O
glucose	B-Simple_chemical
metabolism	O
could	O
affect	O
death	O
receptor	O
-	O
mediated	O
apoptosis	O
in	O
human	B-Organism
tumor	B-Cell
cells	I-Cell
of	O
diverse	O
origin	O
.	O

We	O
show	O
that	O
both	O
substitution	O
of	O
glucose	B-Simple_chemical
for	O
pyruvate	B-Simple_chemical
and	O
treatment	O
with	O
2	B-Simple_chemical
-	I-Simple_chemical
deoxyglucose	I-Simple_chemical
enhanced	O
apoptosis	O
induced	O
by	O
tumor	B-Gene_or_gene_product
necrosis	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	I-Gene_or_gene_product
TNF	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
,	O
CD95	B-Gene_or_gene_product
agonistic	O
antibody	O
,	O
and	O
TNF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
related	I-Gene_or_gene_product
apoptosis	I-Gene_or_gene_product
-	I-Gene_or_gene_product
inducing	I-Gene_or_gene_product
ligand	I-Gene_or_gene_product
(	O
TRAIL	B-Gene_or_gene_product
)	O
.	O

Inhibition	O
of	O
glucose	B-Simple_chemical
metabolism	O
enhanced	O
killing	O
of	O
myeloid	B-Cell
leukemia	I-Cell
U937	I-Cell
,	O
cervical	B-Cell
carcinoma	I-Cell
HeLa	I-Cell
,	O
and	O
breast	B-Cell
carcinoma	I-Cell
MCF	I-Cell
-	I-Cell
7	I-Cell
cells	I-Cell
upon	O
death	O
receptor	O
ligation	O
.	O

Caspase	B-Gene_or_gene_product
activation	O
,	O
mitochondrial	B-Cellular_component
depolarization	O
,	O
and	O
cytochrome	B-Gene_or_gene_product
c	I-Gene_or_gene_product
release	O
were	O
increased	O
under	O
these	O
conditions	O
.	O

Glucose	B-Simple_chemical
deprivation	O
-	O
mediated	O
sensitization	O
to	O
apoptosis	O
was	O
prevented	O
in	O
MCF	B-Cell
-	I-Cell
7	I-Cell
cells	I-Cell
overexpressing	O
BCL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
.	O

Interestingly	O
,	O
the	O
human	B-Organism
B	B-Cell
-	I-Cell
lymphoblastoid	I-Cell
cell	I-Cell
line	I-Cell
SKW6	I-Cell
.	I-Cell
4	I-Cell
,	O
a	O
prototype	O
for	O
mitochondria	B-Cellular_component
-	O
independent	O
death	O
receptor	O
-	O
induced	O
apoptosis	O
,	O
was	O
also	O
sensitized	O
to	O
anti	O
-	O
CD95	B-Gene_or_gene_product
and	O
TRAIL	B-Gene_or_gene_product
-	O
induced	O
apoptosis	O
under	O
glucose	B-Simple_chemical
-	O
free	O
conditions	O
.	O

Changes	O
in	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
FLIP	I-Gene_or_gene_product
(	I-Gene_or_gene_product
L	I-Gene_or_gene_product
)	I-Gene_or_gene_product
and	O
cFLIPs	B-Gene_or_gene_product
levels	O
were	O
observed	O
in	O
some	O
but	O
not	O
all	O
the	O
cell	B-Cell
lines	I-Cell
studied	O
following	O
glucose	B-Simple_chemical
deprivation	O
.	O

Glucose	B-Simple_chemical
deprivation	O
enhanced	O
death	O
receptor	O
-	O
triggered	O
formation	O
of	O
death	B-Gene_or_gene_product
-	I-Gene_or_gene_product
inducing	I-Gene_or_gene_product
signaling	I-Gene_or_gene_product
complex	I-Gene_or_gene_product
and	O
early	O
processing	O
of	O
procaspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
.	O

Altogether	O
,	O
these	O
results	O
suggest	O
that	O
the	O
glycolytic	O
pathway	O
may	O
be	O
an	O
important	O
target	O
for	O
therapeutic	O
intervention	O
to	O
sensitize	O
tumor	B-Cell
cells	I-Cell
to	O
selectively	O
toxic	O
soluble	O
death	O
ligands	O
or	O
death	O
ligand	O
-	O
expressing	O
cells	B-Cell
of	O
the	O
immune	B-Anatomical_system
system	I-Anatomical_system
by	O
facilitating	O
the	O
activation	O
of	O
initiator	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
.	O

Arteriogenesis	O
:	O
the	O
development	O
and	O
growth	O
of	O
collateral	B-Multi-tissue_structure
arteries	I-Multi-tissue_structure
.	O

In	O
patients	B-Organism
with	O
atherosclerotic	O
vascular	B-Multi-tissue_structure
diseases	O
,	O
collateral	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
bypassing	O
major	O
arterial	B-Pathological_formation
obstructions	I-Pathological_formation
have	O
frequently	O
been	O
observed	O
.	O

This	O
may	O
explain	O
why	O
some	O
patients	B-Organism
remain	O
without	O
symptoms	O
or	O
signs	O
of	O
ischemia	O
.	O

The	O
term	O
"	O
arteriogenesis	O
"	O
was	O
introduced	O
to	O
differentiate	O
the	O
formation	O
of	O
collateral	B-Multi-tissue_structure
arteries	I-Multi-tissue_structure
from	O
angiogenesis	O
,	O
which	O
mainly	O
occurs	O
in	O
the	O
ischemic	O
,	O
collateral	O
flow	O
-	O
dependent	O
tissue	B-Tissue
.	O

Many	O
observations	O
in	O
various	O
animal	O
models	O
and	O
humans	B-Organism
support	O
that	O
the	O
remodeling	O
of	O
preexisting	O
collateral	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
is	O
the	O
mechanism	O
of	O
collateral	B-Multi-tissue_structure
artery	I-Multi-tissue_structure
formation	O
.	O

This	O
remodeling	O
process	O
seems	O
to	O
be	O
mainly	O
flow	O
-	O
mediated	O
.	O

It	O
involves	O
endothelial	B-Cell
cell	I-Cell
activation	O
,	O
basal	B-Cellular_component
membrane	I-Cellular_component
degradation	O
,	O
leukocyte	B-Cell
invasion	O
,	O
proliferation	O
of	O
vascular	B-Cell
cells	I-Cell
,	O
neointima	B-Tissue
formation	O
(	O
in	O
most	O
species	O
studied	O
)	O
,	O
and	O
changes	O
of	O
the	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
.	O

The	O
contribution	O
of	O
ischemia	O
to	O
arteriogenesis	O
is	O
still	O
unclear	O
,	O
but	O
arteriogenesis	O
clearly	O
can	O
occur	O
in	O
the	O
absence	O
of	O
any	O
significant	O
ischemia	O
.	O

It	O
is	O
questionable	O
,	O
whether	O
collateral	B-Multi-tissue_structure
arteries	I-Multi-tissue_structure
also	O
form	O
de	O
novo	O
in	O
ischemic	O
vascular	B-Multi-tissue_structure
diseases	O
.	O

A	O
better	O
understanding	O
of	O
the	O
mechanisms	O
of	O
arteriogenesis	O
will	O
be	O
important	O
for	O
the	O
design	O
of	O
more	O
effective	O
strategies	O
for	O
the	O
treatment	O
of	O
patients	B-Organism
with	O
ischemic	O
vascular	B-Multi-tissue_structure
diseases	O
.	O

Bee	B-Organism
venom	B-Organism_substance
induces	O
apoptosis	O
and	O
inhibits	O
expression	O
of	O
cyclooxygenase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
mRNA	O
in	O
human	B-Organism
lung	B-Cell
cancer	I-Cell
cell	I-Cell
line	I-Cell
NCI	I-Cell
-	I-Cell
H1299	I-Cell
.	O

To	O
investigate	O
whether	O
bee	B-Organism
venom	B-Organism_substance
(	O
BV	B-Organism_substance
)	O
induces	O
apoptosis	O
,	O
the	O
3	B-Simple_chemical
-	I-Simple_chemical
(	I-Simple_chemical
4	I-Simple_chemical
,	I-Simple_chemical
5	I-Simple_chemical
-	I-Simple_chemical
dimethylthiazol	I-Simple_chemical
-	I-Simple_chemical
2	I-Simple_chemical
-	I-Simple_chemical
yl	I-Simple_chemical
)	I-Simple_chemical
-	I-Simple_chemical
2	I-Simple_chemical
,	I-Simple_chemical
5	I-Simple_chemical
-	I-Simple_chemical
diphenyltetrazolium	I-Simple_chemical
bromide	I-Simple_chemical
assay	O
,	O
terminal	B-Gene_or_gene_product
deoxynucleotidyl	I-Gene_or_gene_product
transferase	I-Gene_or_gene_product
-	O
mediated	O
dUTP	B-Simple_chemical
nick	O
end	O
-	O
labeling	O
assay	O
,	O
4	B-Simple_chemical
,	I-Simple_chemical
6	I-Simple_chemical
-	I-Simple_chemical
diamidino	I-Simple_chemical
-	I-Simple_chemical
2	I-Simple_chemical
-	I-Simple_chemical
phenylindole	I-Simple_chemical
staining	O
,	O
flow	O
cytometric	O
analysis	O
,	O
and	O
DNA	B-Cellular_component
fragmentation	O
assay	O
were	O
performed	O
on	O
NCI	B-Cell
-	I-Cell
H1299	I-Cell
lung	I-Cell
cancer	I-Cell
cells	I-Cell
treated	O
with	O
BV	B-Organism_substance
.	O

Through	O
morphological	O
and	O
biochemical	O
analyses	O
,	O
it	O
was	O
demonstrated	O
that	O
NCI	B-Cell
-	I-Cell
H1299	I-Cell
cells	I-Cell
treated	O
with	O
BV	B-Organism_substance
exhibit	O
several	O
features	O
of	O
apoptosis	O
.	O

In	O
addition	O
,	O
reverse	O
transcription	O
-	O
polymerase	O
chain	O
reaction	O
and	O
prostaglandin	B-Simple_chemical
E	I-Simple_chemical
(	I-Simple_chemical
2	I-Simple_chemical
)	I-Simple_chemical
(	O
PGE	B-Simple_chemical
(	I-Simple_chemical
2	I-Simple_chemical
)	I-Simple_chemical
)	O
immunoassay	O
were	O
performed	O
to	O
verify	O
whether	O
BV	B-Organism_substance
possesses	O
an	O
inhibitory	O
effect	O
on	O
the	O
expression	O
of	O
cyclooxygenase	B-Gene_or_gene_product
(	O
COX	B-Gene_or_gene_product
)	O
and	O
PGE	B-Simple_chemical
(	I-Simple_chemical
2	I-Simple_chemical
)	I-Simple_chemical
synthesis	O
.	O

Expression	O
of	O
COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
mRNA	O
and	O
synthesis	O
of	O
PGE	B-Simple_chemical
(	I-Simple_chemical
2	I-Simple_chemical
)	I-Simple_chemical
were	O
inhibited	O
by	O
BV	B-Organism_substance
.	O

These	O
results	O
suggest	O
the	O
possibility	O
that	O
BV	B-Organism_substance
may	O
exert	O
an	O
anti	O
-	O
tumor	B-Cancer
effect	O
on	O
human	B-Organism
lung	B-Cancer
cancer	I-Cancer
.	O

Possible	O
role	O
of	O
cyclooxygenase	B-Gene_or_gene_product
II	I-Gene_or_gene_product
in	O
the	O
acquisition	O
of	O
ovarian	B-Multi-tissue_structure
luteal	I-Multi-tissue_structure
function	O
in	O
rodents	O
.	O

The	O
development	O
of	O
the	O
corpus	B-Multi-tissue_structure
luteum	I-Multi-tissue_structure
(	O
CL	B-Multi-tissue_structure
)	O
,	O
which	O
involves	O
angiogenesis	O
,	O
is	O
essential	O
for	O
the	O
establishment	O
of	O
early	O
pregnancy	O
.	O

We	O
investigated	O
the	O
roles	O
of	O
the	O
prostaglandin	B-Simple_chemical
synthases	O
cyclooxygenase	B-Gene_or_gene_product
(	I-Gene_or_gene_product
COX	I-Gene_or_gene_product
)	I-Gene_or_gene_product
I	I-Gene_or_gene_product
and	O
COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
II	I-Gene_or_gene_product
in	O
angiogenesis	O
and	O
progesterone	B-Simple_chemical
production	O
in	O
the	O
newly	O
formed	O
CL	B-Multi-tissue_structure
,	O
using	O
inhibitors	O
of	O
the	O
COX	B-Gene_or_gene_product
enzymes	I-Gene_or_gene_product
and	O
the	O
gonadotropin	B-Gene_or_gene_product
-	O
induced	O
pseudopregnant	O
rat	B-Organism
as	O
a	O
model	O
.	O

Injection	O
of	O
indomethacin	B-Simple_chemical
,	O
a	O
nonselective	O
COX	B-Gene_or_gene_product
inhibitor	O
,	O
on	O
the	O
day	O
of	O
ovulation	O
and	O
the	O
following	O
day	O
decreased	O
serum	B-Organism_substance
levels	O
of	O
progesterone	B-Simple_chemical
,	O
as	O
did	O
injection	O
of	O
the	O
selective	O
COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
II	I-Gene_or_gene_product
inhibitor	O
NS	B-Simple_chemical
-	I-Simple_chemical
398	I-Simple_chemical
.	O

In	O
contrast	O
,	O
a	O
selective	O
COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
I	I-Gene_or_gene_product
inhibitor	O
,	O
SC	B-Simple_chemical
-	I-Simple_chemical
560	I-Simple_chemical
,	O
had	O
no	O
effect	O
on	O
serum	B-Organism_substance
progesterone	B-Simple_chemical
concentrations	O
.	O

None	O
of	O
the	O
inhibitors	O
had	O
any	O
effect	O
on	O
the	O
weight	O
of	O
the	O
superovulated	B-Organ
ovaries	I-Organ
or	O
on	O
the	O
synthesis	O
of	O
progesterone	B-Simple_chemical
by	O
cultured	B-Cell
luteal	I-Cell
cells	I-Cell
.	O

To	O
determine	O
whether	O
changes	O
in	O
angiogenesis	O
are	O
responsible	O
for	O
the	O
decrease	O
in	O
progesterone	B-Simple_chemical
synthesis	O
,	O
we	O
measured	O
hemoglobin	B-Gene_or_gene_product
and	O
CD34	B-Gene_or_gene_product
levels	O
in	O
luteinized	O
ovaries	B-Organ
following	O
injection	O
of	O
COX	B-Gene_or_gene_product
inhibitors	O
and	O
measured	O
the	O
relative	O
frequency	O
of	O
cells	B-Cell
positive	O
for	O
platelet	B-Gene_or_gene_product
-	I-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
cell	I-Gene_or_gene_product
adhesion	I-Gene_or_gene_product
molecule	I-Gene_or_gene_product
as	O
a	O
specific	O
marker	O
for	O
endothelial	B-Cell
cells	I-Cell
.	O

All	O
of	O
these	O
parameters	O
were	O
reduced	O
by	O
the	O
COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
II	I-Gene_or_gene_product
inhibitors	O
,	O
suggesting	O
that	O
changes	O
in	O
the	O
vasculature	B-Multi-tissue_structure
are	O
responsible	O
for	O
the	O
decrease	O
in	O
serum	B-Organism_substance
progesterone	B-Simple_chemical
.	O

Histological	O
examination	O
of	O
ovarian	B-Organ
corrosion	O
casts	O
indicated	O
that	O
NS	B-Simple_chemical
-	I-Simple_chemical
398	I-Simple_chemical
inhibited	O
the	O
establishment	O
of	O
luteal	B-Multi-tissue_structure
capillary	I-Multi-tissue_structure
vessels	I-Multi-tissue_structure
following	O
the	O
injection	O
of	O
hCG	B-Gene_or_gene_product
.	O

The	O
results	O
are	O
consistent	O
with	O
the	O
hypothesis	O
that	O
the	O
activity	O
of	O
COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
II	I-Gene_or_gene_product
is	O
associated	O
with	O
the	O
formation	O
of	O
functional	O
CL	B-Multi-tissue_structure
via	O
its	O
stimulation	O
of	O
angiogenesis	O
.	O

Tumor	B-Cancer
angiogenesis	O
modulates	O
leukocyte	B-Cell
-	O
vessel	B-Multi-tissue_structure
wall	I-Multi-tissue_structure
interactions	O
in	O
vivo	O
by	O
reducing	O
endothelial	B-Gene_or_gene_product
adhesion	I-Gene_or_gene_product
molecule	I-Gene_or_gene_product
expression	O
.	O

The	O
expression	O
of	O
endothelial	B-Gene_or_gene_product
cell	I-Gene_or_gene_product
(	I-Gene_or_gene_product
EC	I-Gene_or_gene_product
)	I-Gene_or_gene_product
adhesion	I-Gene_or_gene_product
molecules	I-Gene_or_gene_product
involved	O
in	O
leukocyte	B-Cell
-	O
vessel	B-Multi-tissue_structure
wall	I-Multi-tissue_structure
interactions	O
is	O
suppressed	O
in	O
malignancies	B-Cancer
.	O

In	O
the	O
present	O
study	O
,	O
we	O
investigated	O
in	O
vivo	O
the	O
regulation	O
of	O
leukocyte	B-Cell
-	O
vessel	B-Multi-tissue_structure
wall	I-Multi-tissue_structure
interactions	O
by	O
the	O
presence	O
of	O
a	O
tumor	B-Cancer
.	O

By	O
means	O
of	O
intravital	O
microscopy	O
,	O
tumor	B-Gene_or_gene_product
necrosis	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
-	O
stimulated	O
leukocyte	B-Cell
-	O
vessel	B-Multi-tissue_structure
wall	I-Multi-tissue_structure
interactions	O
were	O
studied	O
in	O
ear	B-Tissue
skin	I-Tissue
microvessels	I-Tissue
of	O
nude	B-Organism
mice	I-Organism
bearing	O
small	O
human	B-Organism
LS174T	B-Cancer
colon	I-Cancer
carcinomas	I-Cancer
and	O
in	O
C57Bl	B-Organism
/	I-Organism
6	I-Organism
mice	I-Organism
bearing	O
murine	B-Organism
B16F10	B-Cancer
melanomas	I-Cancer
.	O

Leukocyte	B-Cell
-	O
vessel	B-Multi-tissue_structure
wall	I-Multi-tissue_structure
interactions	O
were	O
studied	O
both	O
within	O
and	O
outside	O
small	O
tumors	B-Cancer
growing	O
in	O
the	O
ear	B-Organ
,	O
and	O
in	O
ear	B-Tissue
microvessels	I-Tissue
of	O
mice	B-Organism
with	O
a	O
large	O
tumor	B-Cancer
growing	O
on	O
their	O
flank	B-Organism_subdivision
.	O

Tumor	B-Cancer
-	O
free	O
mice	B-Organism
were	O
used	O
as	O
controls	O
.	O

Compared	O
with	O
values	O
measured	O
at	O
the	O
edge	O
of	O
the	O
ear	B-Organ
and	O
in	O
the	O
contralateral	O
ear	B-Organ
,	O
leukocyte	B-Cell
adhesion	O
was	O
found	O
to	O
be	O
diminished	O
significantly	O
in	O
vessels	B-Tissue
inside	O
the	O
ear	B-Cancer
tumor	I-Cancer
in	O
both	O
mouse	B-Organism
models	O
.	O

This	O
reduction	O
disappeared	O
with	O
increasing	O
distance	O
from	O
the	O
tumor	B-Cancer
.	O

Surprisingly	O
,	O
the	O
level	O
of	O
leukocyte	B-Cell
adhesion	O
in	O
ear	B-Multi-tissue_structure
venules	I-Multi-tissue_structure
of	O
mice	B-Organism
with	O
a	O
large	O
flank	B-Cancer
tumor	I-Cancer
was	O
also	O
reduced	O
significantly	O
.	O

Leukocyte	B-Cell
rolling	O
,	O
i	O
.	O
e	O
.	O
,	O
the	O
step	O
preceding	O
adhesion	O
,	O
was	O
not	O
influenced	O
by	O
the	O
presence	O
of	O
a	O
tumor	B-Cancer
in	O
nude	B-Organism
mice	I-Organism
,	O
but	O
was	O
down	O
-	O
regulated	O
in	O
immune	O
-	O
competent	O
C57Bl	B-Organism
/	I-Organism
6	I-Organism
mice	I-Organism
.	O

Treatment	O
of	O
mice	B-Organism
bearing	O
a	O
small	O
ear	B-Cancer
tumor	I-Cancer
with	O
a	O
humanized	O
antivascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
antibody	I-Gene_or_gene_product
prevented	O
the	O
down	O
-	O
regulation	O
of	O
leukocyte	B-Cell
-	O
vessel	B-Multi-tissue_structure
wall	I-Multi-tissue_structure
interactions	O
inside	O
the	O
tumor	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
compared	O
with	O
the	O
nontreated	O
group	O
.	O

Fluorescence	O
-	O
activated	O
cell	B-Cell
sorter	O
analysis	O
showed	O
that	O
isolated	O
tumor	B-Cell
ECs	I-Cell
have	O
suppressed	O
levels	O
of	O
intercellular	B-Gene_or_gene_product
adhesion	I-Gene_or_gene_product
molecule	I-Gene_or_gene_product
1	I-Gene_or_gene_product
as	O
compared	O
with	O
ECs	B-Cell
from	O
normal	O
mouse	B-Organism
tissues	B-Tissue
.	O

In	O
cultured	O
b	B-Cell
.	I-Cell
END5	I-Cell
cells	I-Cell
the	O
tumor	B-Gene_or_gene_product
necrosis	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
-	O
induced	O
up	O
-	O
regulation	O
of	O
intercellular	B-Gene_or_gene_product
adhesion	I-Gene_or_gene_product
molecule	I-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
vascular	B-Gene_or_gene_product
cell	I-Gene_or_gene_product
adhesion	I-Gene_or_gene_product
molecule	I-Gene_or_gene_product
1	I-Gene_or_gene_product
was	O
reduced	O
in	O
ECs	B-Cell
that	O
were	O
preincubated	O
with	O
basic	B-Gene_or_gene_product
fibroblast	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
or	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
.	O

The	O
current	O
results	O
may	O
have	O
an	O
impact	O
on	O
the	O
effectiveness	O
of	O
clinical	O
immunotherapeutic	O
treatment	O
protocols	O
,	O
because	O
immune	B-Cell
effector	I-Cell
cells	I-Cell
may	O
not	O
be	O
able	O
to	O
enter	O
tumor	B-Tissue
tissue	I-Tissue
.	O

[	O
Effects	O
of	O
Epstein	B-Organism
-	I-Organism
Barr	I-Organism
virus	I-Organism
latent	B-Gene_or_gene_product
membrane	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
EBV	B-Organism
-	O
LMP1	B-Gene_or_gene_product
)	O
on	O
related	O
factors	O
of	O
metastasis	O
of	O
nasopharyngeal	B-Cell
carcinoma	I-Cell
cell	I-Cell
line	I-Cell
CNE1	I-Cell
]	O
.	O

BACKGROUND	O
&	O
#	O
38	O
;	O
OBJECTIVE	O
:	O
It	O
has	O
been	O
proved	O
that	O
Epstein	B-Organism
-	I-Organism
Barr	I-Organism
virus	I-Organism
(	O
EBV	B-Organism
)	O
latent	B-Gene_or_gene_product
membrane	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
EBV	B-Organism
-	O
LMP1	B-Gene_or_gene_product
)	O
can	O
induce	O
the	O
expression	O
of	O
matrix	B-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
(	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
)	O
.	O

This	O
study	O
was	O
designed	O
to	O
investigate	O
the	O
effect	O
of	O
EBV	B-Organism
-	O
LMP1	B-Gene_or_gene_product
on	O
related	O
factors	O
of	O
metastasis	O
of	O
nasopharyngeal	B-Cell
carcinoma	I-Cell
cell	I-Cell
line	I-Cell
CNE1	I-Cell
.	O

METHODS	O
:	O
Expression	O
of	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
was	O
studied	O
in	O
human	B-Organism
NPC	B-Cell
cell	I-Cell
lines	I-Cell
cultured	O
in	O
vitro	O
:	O
CNE1	B-Cell
(	O
well	O
differentiated	O
cell	B-Cell
line	I-Cell
of	O
NPC	B-Cell
)	O
and	O
CNE1	B-Cell
-	I-Cell
GL	I-Cell
(	O
CNE1	B-Cell
cell	I-Cell
line	I-Cell
transfected	O
with	O
an	O
eukaryotic	O
LMP1	B-Gene_or_gene_product
-	O
expression	O
plasmid	O
)	O
by	O
SP	O
immunohistochemistry	O
and	O
Western	O
blot	O
analysis	O
.	O

Cell	B-Cell
-	O
matrix	B-Cellular_component
adhesion	O
assay	O
was	O
used	O
to	O
study	O
the	O
adhesive	O
ability	O
of	O
CNE1	B-Cell
-	I-Cell
GL	I-Cell
cells	I-Cell
.	O

The	O
effects	O
of	O
LMP1	B-Gene_or_gene_product
on	O
the	O
invasion	O
and	O
migration	O
of	O
CNE1	B-Cell
cells	I-Cell
were	O
investigated	O
by	O
transwell	O
methods	O
.	O

RESULTS	O
:	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
was	O
expressed	O
in	O
both	O
cell	B-Cell
lines	I-Cell
but	O
the	O
intensity	O
of	O
the	O
staining	O
was	O
different	O
.	O

The	O
positive	O
rates	O
of	O
expression	O
of	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
in	O
CNE1	B-Cell
and	O
CNE1	B-Cell
-	I-Cell
GL	I-Cell
cells	I-Cell
were	O
30	O
.	O
2	O
%	O
and	O
98	O
.	O
2	O
%	O
,	O
respectively	O
(	O
P	O
<	O
0	O
.	O
05	O
)	O
.	O

The	O
increased	O
expression	O
of	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
was	O
also	O
shown	O
in	O
CNE1	B-Cell
-	I-Cell
GL	I-Cell
cells	I-Cell
by	O
Western	O
blot	O
analysis	O
.	O

Cell	B-Cell
-	O
matrix	B-Cellular_component
adhesion	O
assay	O
showed	O
that	O
the	O
adhesive	O
ability	O
of	O
CNE1	B-Cell
-	I-Cell
GL	I-Cell
with	O
the	O
matrix	B-Cellular_component
(	O
mean	O
A	O
value	O
:	O
1	O
.	O
2508	O
+	O
/	O
-	O
0	O
.	O
0711	O
)	O
was	O
higher	O
than	O
that	O
of	O
CNE1	B-Cell
cell	I-Cell
(	O
mean	O
A	O
value	O
:	O
0	O
.	O
9519	O
+	O
/	O
-	O
0	O
.	O
068	O
)	O
(	O
P	O
<	O
0	O
.	O
001	O
)	O
.	O

Invasion	O
assay	O
and	O
migration	O
assay	O
showed	O
that	O
the	O
invasion	O
and	O
migration	O
of	O
CNE1	B-Cell
-	I-Cell
GL	I-Cell
cell	I-Cell
were	O
higher	O
than	O
those	O
of	O
CNE1	B-Cell
cells	I-Cell
(	O
P	O
<	O
0	O
.	O
01	O
)	O
.	O

CONCLUSION	O
:	O
The	O
transfection	O
of	O
LMP1	B-Gene_or_gene_product
can	O
increase	O
the	O
expression	O
of	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
in	O
CNE1	B-Cell
cells	I-Cell
.	O

Abilities	O
of	O
adhesion	O
,	O
migration	O
,	O
and	O
invasion	O
of	O
CNE1	B-Cell
cell	I-Cell
were	O
induced	O
by	O
LMP1	B-Gene_or_gene_product
.	O

It	O
is	O
suggested	O
that	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
may	O
have	O
a	O
role	O
in	O
the	O
LMP1	B-Gene_or_gene_product
-	O
induced	O
acceleration	O
of	O
invasion	O
and	O
metastasis	O
of	O
NPC	B-Cell
cells	I-Cell
.	O

Truncated	O
galectin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
inhibits	O
tumor	B-Cancer
growth	O
and	O
metastasis	O
in	O
orthotopic	O
nude	B-Organism
mouse	I-Organism
model	O
of	O
human	B-Organism
breast	B-Cancer
cancer	I-Cancer
.	O

PURPOSE	O
:	O
The	O
goal	O
of	O
this	O
research	O
was	O
to	O
evaluate	O
a	O
potential	O
therapeutic	O
agent	O
for	O
breast	B-Cancer
cancer	I-Cancer
based	O
on	O
galectin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
that	O
has	O
been	O
implicated	O
in	O
tumorigenicity	O
and	O
metastasis	O
of	O
breast	B-Cancer
cancer	I-Cancer
.	O

The	O
hypothesis	O
was	O
that	O
therapy	O
with	O
NH	O
(	O
2	O
)	O
-	O
terminally	O
truncated	O
form	O
of	O
galectin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
(	O
galectin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3C	I-Gene_or_gene_product
)	O
will	O
be	O
efficacious	O
for	O
reduction	O
in	O
tumor	B-Cancer
growth	O
and	O
for	O
inhibition	O
of	O
metastases	O
.	O

EXPERIMENTAL	O
DESIGN	O
:	O
Recombinant	O
human	B-Organism
galectin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
was	O
produced	O
in	O
Escherichia	B-Organism
coli	I-Organism
from	O
which	O
galectin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3C	I-Gene_or_gene_product
was	O
derived	O
by	O
collagenase	B-Gene_or_gene_product
enzyme	O
digestion	O
.	O

Toxicity	O
,	O
pharmacokinetic	O
,	O
and	O
organ	B-Organ
biodistribution	O
studies	O
were	O
performed	O
in	O
nude	B-Organism
mice	I-Organism
.	O

For	O
efficacy	O
studies	O
,	O
nude	B-Organism
mice	I-Organism
bearing	O
orthotopically	O
implanted	O
tumors	B-Cancer
derived	O
from	O
breast	B-Cell
cancer	I-Cell
cell	I-Cell
line	I-Cell
MDA	I-Cell
-	I-Cell
MB	I-Cell
-	I-Cell
435	I-Cell
were	O
treated	O
with	O
galectin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3C	I-Gene_or_gene_product
or	O
a	O
vehicle	O
control	O
i	O
.	O
m	O
.	O
twice	O
daily	O
for	O
90	O
days	O
.	O

RESULTS	O
:	O
The	O
maximum	O
tolerated	O
dose	O
of	O
galectin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3C	I-Gene_or_gene_product
in	O
nude	B-Organism
mice	I-Organism
was	O
determined	O
to	O
be	O
>	O
125	O
mg	O
/	O
kg	O
without	O
overt	O
adverse	O
effects	O
.	O

The	O
elimination	O
half	O
-	O
life	O
when	O
administered	O
i	O
.	O
m	O
.	O
was	O
found	O
to	O
be	O
3	O
.	O
0	O
h	O
in	O
the	O
serum	B-Organism_substance
and	O
4	O
.	O
3	O
h	O
in	O
the	O
cellular	B-Cell
fraction	O
of	O
the	O
blood	B-Organism_substance
.	O

Organ	O
biodistribution	O
studies	O
revealed	O
that	O
galectin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3C	I-Gene_or_gene_product
localized	O
in	O
the	O
liver	B-Organ
,	O
kidneys	B-Organ
,	O
and	O
spleen	B-Organ
but	O
not	O
in	O
the	O
heart	B-Organ
or	O
lungs	B-Organ
.	O

We	O
found	O
that	O
the	O
mean	O
tumor	B-Cancer
volumes	O
and	O
weights	O
were	O
statistically	O
significantly	O
less	O
in	O
mice	B-Organism
treated	O
with	O
galectin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3C	I-Gene_or_gene_product
compared	O
with	O
control	O
mice	B-Organism
,	O
and	O
that	O
fewer	O
numbers	O
of	O
mice	B-Organism
exhibited	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastases	O
in	O
the	O
treated	O
group	O
compared	O
with	O
the	O
control	O
group	O
.	O

CONCLUSIONS	O
:	O
Galectin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3C	I-Gene_or_gene_product
is	O
not	O
overtly	O
toxic	O
,	O
and	O
is	O
efficacious	O
in	O
reducing	O
metastases	O
and	O
tumor	B-Cancer
volumes	O
and	O
weights	O
in	O
primary	O
tumors	B-Cancer
in	O
an	O
orthotopic	O
nude	B-Organism
mouse	I-Organism
model	O
of	O
human	B-Organism
breast	B-Cancer
cancer	I-Cancer
.	O

Prognostic	O
value	O
of	O
p53	B-Gene_or_gene_product
protein	O
expression	O
and	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
expression	O
in	O
resected	O
squamous	B-Cancer
cell	I-Cancer
carcinoma	I-Cancer
of	O
the	O
esophagus	B-Organ
.	O

The	O
most	O
common	O
genetic	O
alterations	O
found	O
in	O
a	O
wide	O
variety	O
of	O
cancers	B-Cancer
are	O
p53	B-Gene_or_gene_product
tumor	B-Cancer
suppressor	O
gene	O
mutations	O
.	O

p53	B-Gene_or_gene_product
appears	O
to	O
be	O
a	O
nuclear	O
transcription	O
factor	O
that	O
plays	O
a	O
role	O
in	O
the	O
control	O
of	O
cell	B-Cell
proliferation	O
,	O
apoptosis	O
,	O
and	O
the	O
maintenance	O
of	O
genetic	O
stability	O
.	O

Angiogenesis	O
is	O
a	O
critical	O
process	O
in	O
solid	B-Cancer
tumor	I-Cancer
growth	O
and	O
metastasis	O
.	O

Vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
,	O
a	O
recently	O
identified	O
growth	O
factor	O
with	O
significant	O
angiogenic	O
properties	O
,	O
may	O
be	O
a	O
major	O
tumor	B-Cancer
angiogenesis	O
regulator	O
.	O

Few	O
studies	O
have	O
investigated	O
the	O
association	O
between	O
p53	B-Gene_or_gene_product
and	O
VEGF	B-Gene_or_gene_product
expressions	O
and	O
prognosis	O
in	O
esophageal	B-Cancer
carcinoma	I-Cancer
.	O

Forty	O
-	O
seven	O
specimens	O
resected	O
from	O
patients	B-Organism
with	O
stage	O
II	O
and	O
III	O
squamous	B-Cancer
cell	I-Cancer
carcinoma	I-Cancer
(	O
SCC	B-Cancer
)	O
of	O
the	O
esophagus	B-Organ
were	O
studied	O
using	O
immunohistochemical	O
staining	O
.	O

VEGF	B-Gene_or_gene_product
and	O
p53	B-Gene_or_gene_product
expressions	O
were	O
observed	O
in	O
40	O
%	O
and	O
53	O
%	O
of	O
the	O
tumors	B-Cancer
,	O
respectively	O
.	O

The	O
p53	B-Gene_or_gene_product
and	O
VEGF	B-Gene_or_gene_product
staining	O
statuses	O
were	O
coincident	O
in	O
only	O
21	O
%	O
of	O
the	O
tumors	B-Cancer
,	O
and	O
no	O
significant	O
correlation	O
was	O
found	O
between	O
p53	B-Gene_or_gene_product
and	O
VEGF	B-Gene_or_gene_product
statuses	O
.	O

No	O
clinicopathologic	O
factors	O
were	O
significantly	O
correlated	O
with	O
p53	B-Gene_or_gene_product
or	O
VEGF	B-Gene_or_gene_product
expression	O
.	O

No	O
significant	O
association	O
between	O
p53	B-Gene_or_gene_product
and	O
VEGF	B-Gene_or_gene_product
expressions	O
and	O
poor	O
prognosis	O
was	O
found	O
.	O

In	O
conclusion	O
,	O
p53	B-Gene_or_gene_product
and	O
VEGF	B-Gene_or_gene_product
were	O
not	O
correlated	O
with	O
prognosis	O
in	O
patients	B-Organism
with	O
stage	O
II	O
and	O
III	O
SCC	B-Cancer
of	O
the	O
esophagus	B-Organ
.	O

Therapeutic	O
targeting	O
of	O
the	O
survivin	B-Gene_or_gene_product
pathway	O
in	O
cancer	B-Cancer
:	O
initiation	O
of	O
mitochondrial	B-Cellular_component
apoptosis	O
and	O
suppression	O
of	O
tumor	B-Cancer
-	O
associated	O
angiogenesis	O
.	O

PURPOSE	O
:	O
Molecular	O
antagonists	O
of	O
the	O
inhibitor	O
of	O
apoptosis	O
protein	O
survivin	B-Gene_or_gene_product
have	O
shown	O
promise	O
as	O
novel	O
anticancer	B-Cancer
strategies	O
for	O
triggering	O
tumor	B-Cell
cell	I-Cell
apoptosis	O
,	O
dysregulating	O
mitotic	O
progression	O
,	O
and	O
inhibiting	O
tumor	B-Cancer
growth	O
in	O
preclinical	O
models	O
.	O

However	O
,	O
how	O
survivin	B-Gene_or_gene_product
couples	O
to	O
the	O
cell	B-Cell
death	O
machinery	O
has	O
remained	O
elusive	O
,	O
and	O
the	O
relevant	O
cellular	B-Cell
targets	O
of	O
survivin	B-Gene_or_gene_product
antagonists	O
have	O
not	O
been	O
completely	O
elucidated	O
.	O

Experimental	O
Design	O
:	O
Human	B-Organism
umbilical	B-Cell
vein	I-Cell
and	O
dermal	B-Cell
microvascular	I-Cell
endothelial	I-Cell
cells	I-Cell
were	O
infected	O
with	O
replication	O
-	O
deficient	O
adenoviruses	B-Organism
encoding	O
survivin	B-Gene_or_gene_product
(	O
pAd	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Survivin	I-Gene_or_gene_product
)	O
,	O
green	B-Gene_or_gene_product
fluorescent	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
(	O
pAd	B-Gene_or_gene_product
-	I-Gene_or_gene_product
GFP	I-Gene_or_gene_product
)	O
,	O
or	O
a	O
phosphorylation	O
-	O
defective	O
survivin	B-Gene_or_gene_product
Thr	O
(	O
34	O
)	O
-	O
-	O
>	O
Ala	O
(	O
pAd	B-Gene_or_gene_product
-	I-Gene_or_gene_product
T34A	I-Gene_or_gene_product
)	O
dominant	O
negative	O
mutant	O
.	O

The	O
effect	O
of	O
wild	O
-	O
type	O
or	O
mutant	O
survivin	B-Gene_or_gene_product
was	O
investigated	O
on	O
capillary	B-Multi-tissue_structure
network	I-Multi-tissue_structure
stability	O
,	O
endothelial	B-Cell
cell	I-Cell
viability	O
,	O
and	O
caspase	B-Gene_or_gene_product
activation	O
in	O
vitro	O
and	O
on	O
kinetics	O
of	O
tumor	B-Cancer
growth	O
and	O
development	O
of	O
angiogenesis	O
in	O
a	O
breast	B-Cancer
cancer	I-Cancer
xenograft	I-Cancer
model	O
in	O
vivo	O
.	O

The	O
cell	B-Cell
death	O
pathway	O
initiated	O
by	O
survivin	B-Gene_or_gene_product
targeting	O
was	O
mapped	O
with	O
respect	O
to	O
cytochrome	B-Gene_or_gene_product
c	I-Gene_or_gene_product
release	O
,	O
changes	O
in	O
mitochondrial	B-Cellular_component
transmembrane	O
potential	O
,	O
and	O
apoptosome	O
requirements	O
using	O
mouse	B-Organism
embryonic	B-Cell
fibroblasts	I-Cell
deficient	O
in	O
Apaf	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
or	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
.	O

RESULTS	O
:	O
Adenoviral	B-Organism
transduction	O
of	O
endothelial	B-Cell
cells	I-Cell
with	O
pAd	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Survivin	I-Gene_or_gene_product
inhibited	O
growth	O
factor	O
deprivation	O
-	O
or	O
ceramide	B-Simple_chemical
-	O
induced	O
apoptosis	O
,	O
reduced	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
and	O
-	B-Gene_or_gene_product
7	I-Gene_or_gene_product
generation	O
,	O
and	O
stabilized	O
three	O
-	O
dimensional	O
capillary	B-Multi-tissue_structure
networks	I-Multi-tissue_structure
in	O
vitro	O
.	O

Conversely	O
,	O
expression	O
of	O
pAd	B-Gene_or_gene_product
-	I-Gene_or_gene_product
T34A	I-Gene_or_gene_product
caused	O
apoptosis	O
in	O
umbilical	B-Cell
vein	I-Cell
and	O
dermal	B-Cell
microvascular	I-Cell
endothelial	I-Cell
cells	I-Cell
and	O
resulted	O
in	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
activity	O
.	O

Cell	B-Cell
death	O
induced	O
by	O
survivin	B-Gene_or_gene_product
targeting	O
exhibited	O
the	O
hallmarks	O
of	O
mitochondrial	B-Cellular_component
-	O
dependent	O
apoptosis	O
with	O
release	O
of	O
cytochrome	B-Gene_or_gene_product
c	I-Gene_or_gene_product
and	O
loss	O
of	O
mitochondrial	B-Cellular_component
transmembrane	O
potential	O
and	O
was	O
suppressed	O
in	O
Apaf	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
or	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
knockout	O
mouse	B-Organism
embryonic	B-Cell
fibroblasts	I-Cell
.	O

When	O
injected	O
in	O
human	B-Organism
breast	B-Cancer
cancer	I-Cancer
xenografts	I-Cancer
,	O
pAd	B-Gene_or_gene_product
-	I-Gene_or_gene_product
T34A	I-Gene_or_gene_product
inhibited	O
growth	O
of	O
established	O
tumors	B-Cancer
and	O
triggered	O
tumor	B-Cell
cell	I-Cell
apoptosis	O
in	O
vivo	O
.	O

This	O
was	O
associated	O
with	O
a	O
approximately	O
60	O
%	O
reduction	O
in	O
tumor	B-Cancer
-	O
derived	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
by	O
quantitative	O
morphometry	O
of	O
CD31	B-Gene_or_gene_product
-	O
stained	O
tumor	B-Cancer
areas	O
,	O
and	O
appearance	O
of	O
endothelial	B-Cell
cell	I-Cell
apoptosis	O
by	O
internucleosomal	O
DNA	B-Cellular_component
fragmentation	O
in	O
vivo	O
.	O

CONCLUSIONS	O
:	O
Survivin	B-Gene_or_gene_product
functions	O
as	O
a	O
novel	O
upstream	O
regulator	O
of	O
mitochondrial	B-Cellular_component
-	O
dependent	O
apoptosis	O
,	O
and	O
molecular	O
targeting	O
of	O
this	O
pathway	O
results	O
in	O
anticancer	B-Cancer
activity	O
via	O
a	O
dual	O
mechanism	O
of	O
induction	O
of	O
tumor	B-Cell
cell	I-Cell
apoptosis	O
and	O
suppression	O
of	O
angiogenesis	O
.	O

The	O
role	O
of	O
organ	B-Organ
vascularization	O
and	O
lipoplex	B-Simple_chemical
-	O
serum	B-Organism_substance
initial	O
contact	O
in	O
intravenous	B-Immaterial_anatomical_entity
murine	B-Organism
lipofection	O
.	O

Following	O
intravenous	B-Immaterial_anatomical_entity
administration	O
of	O
cationic	O
lipid	O
-	O
DNA	B-Cellular_component
complexes	O
(	O
lipoplexes	B-Simple_chemical
)	O
into	O
mice	B-Organism
,	O
transfection	O
(	O
lipofection	O
)	O
occurs	O
predominantly	O
in	O
the	O
lungs	B-Organ
.	O

This	O
was	O
attributed	O
to	O
high	O
entrapment	O
of	O
lipoplexes	B-Simple_chemical
in	O
the	O
extended	O
lung	B-Multi-tissue_structure
vascular	I-Multi-tissue_structure
tree	I-Multi-tissue_structure
.	O

To	O
determine	O
whether	O
lipofection	O
in	O
other	O
organs	B-Organ
could	O
be	O
enhanced	O
by	O
increasing	O
the	O
degree	O
of	O
vascularization	O
,	O
we	O
used	O
a	O
transgenic	O
mouse	B-Organism
model	O
with	O
tissue	B-Tissue
-	O
specific	O
angiogenesis	O
in	O
liver	B-Organ
.	O

Tail	B-Multi-tissue_structure
vein	I-Multi-tissue_structure
injection	O
of	O
N	B-Simple_chemical
-	I-Simple_chemical
(	I-Simple_chemical
1	I-Simple_chemical
-	I-Simple_chemical
(	I-Simple_chemical
2	I-Simple_chemical
,	I-Simple_chemical
3	I-Simple_chemical
-	I-Simple_chemical
dioleoyloxy	I-Simple_chemical
)	I-Simple_chemical
propyl	I-Simple_chemical
)	I-Simple_chemical
-	I-Simple_chemical
N	I-Simple_chemical
,	I-Simple_chemical
N	I-Simple_chemical
,	I-Simple_chemical
N	I-Simple_chemical
-	I-Simple_chemical
trimethylammonium	I-Simple_chemical
chloride	I-Simple_chemical
(	I-Simple_chemical
DOTAP	I-Simple_chemical
)	I-Simple_chemical
/	I-Simple_chemical
cholesterol	I-Simple_chemical
lipoplexes	I-Simple_chemical
resulted	O
in	O
increased	O
lipoplex	B-Simple_chemical
entrapment	O
in	O
hypervascularized	O
liver	B-Organ
but	O
did	O
not	O
boost	O
luciferase	B-Gene_or_gene_product
expression	O
,	O
suggesting	O
that	O
lipoplex	B-Simple_chemical
delivery	O
is	O
not	O
a	O
sufficient	O
condition	O
for	O
efficient	O
organ	B-Organ
lipofection	O
.	O

Because	O
the	O
intravenously	B-Immaterial_anatomical_entity
injected	O
lipoplexes	B-Simple_chemical
migrated	O
within	O
seconds	O
to	O
lungs	B-Organ
,	O
we	O
checked	O
whether	O
the	O
effects	O
of	O
immediate	O
contact	O
with	O
serum	B-Organism_substance
correlate	O
with	O
lung	B-Organ
lipofection	O
efficiency	O
of	O
different	O
DOTAP	B-Simple_chemical
-	O
based	O
formulations	O
.	O

Under	O
conditions	O
mimicking	O
the	O
injection	O
environment	O
,	O
the	O
lipoplex	B-Simple_chemical
-	O
serum	B-Organism_substance
interaction	O
was	O
strongly	O
dependent	O
on	O
helper	O
lipid	B-Simple_chemical
and	O
ionic	O
strength	O
:	O
lipoplexes	B-Simple_chemical
prepared	O
in	O
150	O
mM	O
NaCl	O
or	O
lipoplexes	B-Simple_chemical
with	O
high	O
(	O
greater	O
than	O
33	O
mol	O
%	O
)	O
cholesterol	B-Simple_chemical
were	O
found	O
to	O
aggregate	O
immediately	O
.	O

This	O
aggregation	O
process	O
was	O
irreversible	O
and	O
was	O
inversely	O
correlated	O
with	O
the	O
percentage	O
of	O
lung	B-Cell
cells	I-Cell
that	O
took	O
up	O
lipoplexes	B-Simple_chemical
and	O
with	O
the	O
efficiency	O
of	O
lipofection	O
.	O

No	O
other	O
structural	O
changes	O
in	O
serum	B-Organism_substance
were	O
observed	O
for	O
cholesterol	B-Simple_chemical
-	I-Simple_chemical
based	I-Simple_chemical
lipoplexes	I-Simple_chemical
.	O

Dioleoyl	B-Simple_chemical
phosphatidylethanolamine	I-Simple_chemical
-	I-Simple_chemical
based	I-Simple_chemical
lipoplexes	I-Simple_chemical
were	O
found	O
to	O
give	O
low	O
expression	O
,	O
apparently	O
because	O
of	O
an	O
immediate	O
loss	O
of	O
integrity	O
in	O
serum	B-Organism_substance
,	O
without	O
lipid	O
-	O
DNA	B-Cellular_component
dissociation	O
.	O

Our	O
study	O
suggests	O
that	O
efficient	O
in	O
vivo	O
lipofection	O
is	O
the	O
result	O
of	O
cross	O
-	O
talk	O
between	O
lipoplex	B-Simple_chemical
composition	O
,	O
interaction	O
with	O
serum	B-Organism_substance
,	O
hemodynamics	O
,	O
and	O
target	O
tissue	B-Tissue
"	O
susceptibility	O
"	O
to	O
transfection	O
.	O

Proliferative	O
diabetic	O
retinopathy	O
is	O
associated	O
with	O
a	O
low	O
level	O
of	O
the	O
natural	O
ocular	B-Organ
anti	O
-	O
angiogenic	O
agent	O
pigment	B-Gene_or_gene_product
epithelium	I-Gene_or_gene_product
-	I-Gene_or_gene_product
derived	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
PEDF	B-Gene_or_gene_product
)	O
in	O
aqueous	B-Organism_substance
humor	I-Organism_substance
.	O

a	O
pilot	O
study	O
.	O

Retinopathy	O
is	O
the	O
most	O
common	O
microvascular	B-Tissue
diabetes	O
complication	O
and	O
represents	O
a	O
major	O
threat	O
to	O
the	O
eyesight	O
.	O

The	O
aim	O
of	O
this	O
study	O
was	O
to	O
address	O
the	O
role	O
of	O
pro	O
-	O
and	O
anti	O
-	O
angiogenic	O
molecules	O
in	O
diabetic	O
retinopathy	O
in	O
the	O
aqueous	B-Organism_substance
humor	I-Organism_substance
of	O
the	O
eye	B-Organ
.	O

Aqueous	B-Organism_substance
humor	I-Organism_substance
was	O
collected	O
at	O
cataract	O
surgery	O
from	O
19	O
diabetic	O
patients	B-Organism
and	O
from	O
13	O
age	O
-	O
and	O
sex	O
-	O
matched	O
normoglycemic	O
controls	B-Organism
.	O

Levels	O
of	O
pro	O
-	O
angiogenic	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
and	O
angiogenic	O
inhibitor	O
pigment	B-Gene_or_gene_product
epithelium	I-Gene_or_gene_product
-	I-Gene_or_gene_product
derived	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
PEDF	B-Gene_or_gene_product
)	O
were	O
determined	O
.	O

Angiogenic	O
activity	O
of	O
the	O
aqueous	B-Organism_substance
humor	I-Organism_substance
was	O
quantified	O
by	O
measuring	O
its	O
effect	O
on	O
the	O
migration	O
of	O
capillary	B-Cell
endothelial	I-Cell
cells	I-Cell
.	O

In	O
the	O
aqueous	B-Organism_substance
fluid	I-Organism_substance
,	O
VEGF	B-Gene_or_gene_product
levels	O
were	O
increased	O
in	O
diabetics	O
(	O
mean	O
values	O
:	O
501	O
vs	O
.	O
367	O
pg	O
/	O
ml	O
;	O
p	O
=	O
0	O
.	O
05	O
)	O
,	O
compared	O
to	O
controls	O
.	O

PEDF	B-Gene_or_gene_product
was	O
found	O
to	O
be	O
decreased	O
in	O
diabetics	O
(	O
mean	O
values	O
:	O
2080	O
vs	O
.	O
5780	O
ng	O
/	O
ml	O
;	O
p	O
=	O
0	O
.	O
04	O
)	O
compared	O
to	O
controls	O
.	O

In	O
seven	O
diabetic	O
patients	B-Organism
with	O
proliferative	O
retinopathy	O
,	O
the	O
most	O
profound	O
finding	O
was	O
a	O
significant	O
decrease	O
of	O
the	O
PEDF	B-Gene_or_gene_product
level	O
(	O
mean	O
value	O
:	O
237	O
ng	O
/	O
ml	O
)	O
,	O
whereas	O
VEGF	B-Gene_or_gene_product
levels	O
were	O
comparable	O
to	O
diabetic	O
patients	B-Organism
without	O
proliferation	O
(	O
mean	O
value	O
:	O
3153	O
;	O
p	O
=	O
0	O
.	O
003	O
)	O
.	O

Angiogenic	O
activity	O
in	O
samples	O
of	O
patients	B-Organism
from	O
the	O
control	O
group	O
was	O
generally	O
inhibitory	O
due	O
to	O
PEDF	B-Gene_or_gene_product
,	O
and	O
inhibition	O
was	O
blocked	O
by	O
neutralizing	O
antibodies	O
to	O
PEDF	B-Gene_or_gene_product
.	O

Likewise	O
,	O
in	O
diabetics	O
without	O
proliferation	O
,	O
angiogenic	O
activity	O
was	O
also	O
blocked	O
by	O
antibodies	O
to	O
PEDF	B-Gene_or_gene_product
.	O

We	O
will	O
demonstrate	O
here	O
that	O
the	O
level	O
of	O
the	O
natural	O
ocular	O
anti	O
-	O
angiogenic	O
agent	O
PEDF	B-Gene_or_gene_product
is	O
inversely	O
associated	O
with	O
proliferative	O
retinopathy	O
.	O

PEDF	B-Gene_or_gene_product
is	O
an	O
important	O
negative	O
regulator	O
of	O
angiogenic	O
activity	O
of	O
aqueous	B-Organism_substance
humor	I-Organism_substance
.	O

Our	O
data	O
may	O
have	O
implications	O
for	O
the	O
development	O
of	O
novel	O
regimens	O
for	O
diabetic	O
retinopathy	O
.	O

Expression	O
of	O
osteoprotegerin	B-Gene_or_gene_product
and	O
RANK	B-Gene_or_gene_product
ligand	I-Gene_or_gene_product
in	O
breast	B-Cancer
cancer	I-Cancer
bone	B-Tissue
metastasis	O
.	O

Bone	B-Tissue
destruction	O
is	O
primarily	O
mediated	O
by	O
osteoclastic	B-Cell
bone	B-Tissue
resorption	O
,	O
and	O
cancer	B-Cell
cells	I-Cell
stimulate	O
the	O
formation	O
and	O
activation	O
of	O
osteoclasts	B-Cell
next	O
to	O
metastatic	O
foci	B-Cell
.	O

Accumulating	O
evidences	O
indicate	O
that	O
receptor	B-Gene_or_gene_product
activator	I-Gene_or_gene_product
of	I-Gene_or_gene_product
NF	I-Gene_or_gene_product
-	I-Gene_or_gene_product
kB	I-Gene_or_gene_product
ligand	I-Gene_or_gene_product
(	O
RANKL	B-Gene_or_gene_product
)	O
is	O
the	O
ultimate	O
extracellular	B-Immaterial_anatomical_entity
mediator	O
that	O
stimulates	O
osteoclast	B-Cell
differentiation	O
into	O
mature	O
osteoclasts	B-Cell
.	O

In	O
contrast	O
,	O
osteoprotegerin	B-Gene_or_gene_product
(	O
OPG	B-Gene_or_gene_product
)	O
inhibits	O
osteoclast	B-Cell
development	O
.	O

In	O
order	O
to	O
elucidate	O
a	O
mechanism	O
for	O
cancer	B-Cancer
-	O
induced	O
osteoclastogenesis	O
,	O
cells	B-Cell
from	O
a	O
human	B-Organism
breast	B-Cancer
cancer	I-Cancer
line	O
,	O
MDA	B-Cell
-	I-Cell
MB	I-Cell
-	I-Cell
231	I-Cell
,	O
were	O
directly	O
co	O
-	O
cultured	O
with	O
ST2	B-Cell
,	O
MC3T3	B-Cell
-	I-Cell
E1	I-Cell
,	O
or	O
with	O
primary	O
mouse	B-Organism
calvarial	B-Cell
cells	I-Cell
.	O

Osteoclast	B-Cell
-	I-Cell
like	I-Cell
cells	I-Cell
and	O
tartarate	B-Gene_or_gene_product
resistant	I-Gene_or_gene_product
acid	I-Gene_or_gene_product
phosphatase	I-Gene_or_gene_product
(	O
TRAP	B-Gene_or_gene_product
)	O
activities	O
were	O
then	O
quantitated	O
.	O

We	O
examined	O
these	O
cell	B-Cell
lines	I-Cell
and	O
samples	B-Tissue
from	O
breast	B-Cancer
cancer	I-Cancer
by	O
RT	O
-	O
PCR	O
for	O
the	O
expressions	O
of	O
OPG	B-Gene_or_gene_product
and	O
RANKL	B-Gene_or_gene_product
mRNA	O
.	O

Compared	O
to	O
controls	O
,	O
co	O
-	O
culture	O
of	O
MDA	B-Cell
-	I-Cell
MB	I-Cell
-	I-Cell
231	I-Cell
cells	I-Cell
with	O
stromal	B-Cell
or	O
osteoblastic	B-Cell
cells	I-Cell
induced	O
an	O
increase	O
in	O
number	O
of	O
osteoclasts	B-Cell
and	O
TRAP	B-Gene_or_gene_product
activities	O
.	O

MDA	B-Cell
-	I-Cell
MB	I-Cell
-	I-Cell
231	I-Cell
cells	I-Cell
alone	O
or	O
breast	B-Cancer
cancer	I-Cancer
samples	O
did	O
not	O
express	O
RANKL	B-Gene_or_gene_product
mRNA	O
.	O

However	O
,	O
co	O
-	O
culture	O
of	O
these	O
cancer	B-Cell
cells	I-Cell
with	O
stromal	B-Cell
or	O
osteoblastic	B-Cell
cells	I-Cell
induced	O
RANKL	B-Gene_or_gene_product
mRNA	O
expression	O
and	O
decreased	O
OPG	B-Gene_or_gene_product
mRNA	O
expression	O
.	O

These	O
experiments	O
demonstrate	O
that	O
direct	O
interactions	O
between	O
breast	B-Cancer
cancer	I-Cancer
and	O
stromal	B-Cell
or	O
osteoblastic	B-Cell
cells	I-Cell
induce	O
osteoclastogenesis	O
in	O
vitro	O
through	O
modulating	O
RANKL	B-Gene_or_gene_product
expression	O
.	O

Interleukin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
antagonist	I-Gene_or_gene_product
inhibits	O
ischaemic	O
and	O
excitotoxic	O
neuronal	B-Cell
damage	O
in	O
the	O
rat	B-Organism
.	O

Interleukin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
synthesis	O
in	O
the	O
brain	B-Organ
is	O
stimulated	O
by	O
mechanical	O
injury	O
and	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
mimics	O
some	O
effects	O
of	O
injury	O
,	O
such	O
as	O
gliosis	O
and	O
neovascularization	O
.	O

We	O
report	O
that	O
neuronal	B-Cell
death	O
resulting	O
from	O
focal	O
cerebral	B-Organ
ischaemia	O
(	O
middle	O
cerebral	B-Multi-tissue_structure
artery	I-Multi-tissue_structure
occlusion	O
,	O
24	O
h	O
)	O
is	O
significantly	O
inhibited	O
(	O
by	O
50	O
%	O
)	O
in	O
rats	B-Organism
injected	O
with	O
a	O
recombinant	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
antagonist	I-Gene_or_gene_product
(	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1ra	I-Gene_or_gene_product
,	O
10	O
micrograms	O
,	O
icv	O
30	O
min	O
before	O
and	O
10	O
min	O
after	O
ischaemia	O
)	O
.	O

Excitotoxic	O
damage	O
due	O
to	O
striatal	B-Multi-tissue_structure
infusion	O
of	O
an	O
NMDA	B-Simple_chemical
-	O
receptor	O
agonist	O
(	O
cis	B-Simple_chemical
-	I-Simple_chemical
2	I-Simple_chemical
,	I-Simple_chemical
4	I-Simple_chemical
-	I-Simple_chemical
methanoglutamate	I-Simple_chemical
)	O
was	O
also	O
markedly	O
inhibited	O
(	O
71	O
%	O
)	O
by	O
injection	O
of	O
the	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1ra	I-Gene_or_gene_product
.	O

These	O
data	O
indicate	O
that	O
endogenous	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
is	O
a	O
mediator	O
of	O
ischaemic	O
and	O
excitotoxic	O
brain	B-Organ
damage	O
,	O
and	O
that	O
inhibitors	O
of	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
action	O
may	O
be	O
of	O
therapeutic	O
value	O
in	O
the	O
treatment	O
of	O
acute	O
or	O
chronic	O
neuronal	B-Cell
death	O
.	O

Transcriptional	O
and	O
post	O
-	O
translation	O
regulation	O
of	O
the	O
Tie1	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
by	O
fluid	O
shear	O
stress	O
changes	O
in	O
vascular	B-Cell
endothelial	I-Cell
cells	I-Cell
.	O

The	O
interaction	O
between	O
the	O
vascular	B-Tissue
endothelium	I-Tissue
and	O
hemodynamic	O
forces	O
(	O
and	O
more	O
specifically	O
,	O
fluid	O
shear	O
stress	O
)	O
,	O
induced	O
by	O
the	O
flow	O
of	O
blood	B-Organism_substance
,	O
plays	O
a	O
major	O
role	O
in	O
vascular	B-Multi-tissue_structure
remodeling	O
and	O
in	O
new	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
formation	O
via	O
a	O
process	O
termed	O
arteriogenesis	O
.	O

Tie1	B-Gene_or_gene_product
is	O
an	O
orphan	O
tyrosine	O
kinase	O
receptor	O
expressed	O
almost	O
exclusively	O
in	O
endothelial	B-Cell
cells	I-Cell
and	O
is	O
required	O
for	O
normal	O
vascular	B-Multi-tissue_structure
development	O
and	O
maintenance	O
.	O

The	O
present	O
study	O
demonstrates	O
that	O
Tie1	B-Gene_or_gene_product
expression	O
is	O
rapidly	O
down	O
-	O
regulated	O
in	O
endothelial	B-Cell
cells	I-Cell
exposed	O
to	O
shear	O
stress	O
,	O
and	O
more	O
so	O
to	O
shear	O
stress	O
changes	O
.	O

This	O
down	O
-	O
regulation	O
is	O
accompanied	O
by	O
a	O
rapid	O
cleavage	O
of	O
Tie1	B-Gene_or_gene_product
and	O
binding	O
of	O
the	O
cleaved	O
Tie1	B-Gene_or_gene_product
45	O
kDa	O
endodomain	O
to	O
Tie2	B-Gene_or_gene_product
.	O

The	O
rapid	O
cleavage	O
of	O
Tie1	B-Gene_or_gene_product
is	O
followed	O
by	O
a	O
transcriptional	O
down	O
-	O
regulation	O
in	O
response	O
to	O
shear	O
stress	O
.	O

The	O
activity	O
of	O
the	O
Tie1	B-Gene_or_gene_product
promoter	O
is	O
suppressed	O
by	O
shear	O
stress	O
and	O
by	O
tumor	B-Gene_or_gene_product
necrosis	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
.	O

Shear	O
stress	O
-	O
induced	O
transcriptional	O
suppression	O
of	O
Tie1	B-Gene_or_gene_product
is	O
mediated	O
by	O
a	O
negative	O
shear	O
stress	O
response	O
element	O
,	O
localized	O
in	O
a	O
region	O
of	O
250	O
bp	O
within	O
the	O
promoter	O
.	O

The	O
rapid	O
down	O
-	O
regulation	O
of	O
Tie1	B-Gene_or_gene_product
by	O
shear	O
stress	O
changes	O
and	O
its	O
rapid	O
binding	O
to	O
Tie2	B-Gene_or_gene_product
may	O
be	O
required	O
for	O
destabilization	O
of	O
endothelial	B-Cell
cells	I-Cell
in	O
order	O
to	O
initiate	O
the	O
process	O
of	O
vascular	B-Multi-tissue_structure
restructuring	O
.	O

Therapeutic	O
angiogenesis	O
:	O
a	O
complex	O
problem	O
requiring	O
a	O
sophisticated	O
approach	O
.	O

Blood	B-Organism_substance
and	O
vascular	B-Multi-tissue_structure
disorders	O
underlie	O
a	O
plethora	O
of	O
pathologic	O
conditions	O
and	O
are	O
the	O
single	O
most	O
frequent	O
cause	O
of	O
human	B-Organism
disease	O
.	O

Ischemia	O
,	O
involving	O
restricted	O
blood	B-Organism_substance
flow	O
to	O
tissues	B-Tissue
is	O
the	O
most	O
common	O
consequence	O
of	O
vessel	B-Multi-tissue_structure
dysfunction	O
resulting	O
in	O
the	O
disruption	O
of	O
oxygen	B-Simple_chemical
and	O
nutrient	O
delivery	O
and	O
the	O
accumulation	O
of	O
waste	O
metabolites	O
.	O

Cells	B-Cell
cannot	O
survive	O
extended	O
severe	O
ischemia	O
but	O
may	O
be	O
able	O
to	O
adapt	O
to	O
a	O
moderate	O
condition	O
where	O
diffusion	O
to	O
and	O
from	O
bordering	O
nonischemic	O
regions	O
sustains	O
vital	O
functions	O
.	O

Under	O
this	O
condition	O
,	O
the	O
secondary	O
functions	O
of	O
effected	O
cells	B-Cell
are	O
likely	O
to	O
be	O
impaired	O
,	O
and	O
a	O
new	O
metabolic	O
equilibrium	O
is	O
established	O
,	O
determined	O
by	O
the	O
level	O
of	O
cross	O
-	O
diffusion	O
and	O
degree	O
of	O
hypoxia	O
.	O

In	O
tissues	B-Tissue
with	O
a	O
normally	O
high	O
metabolic	O
turnover	O
such	O
as	O
skeletal	B-Tissue
and	O
cardiac	B-Tissue
muscle	I-Tissue
,	O
even	O
mild	O
ischemia	O
causes	O
hypoxia	O
,	O
acidosis	O
,	O
and	O
depressed	O
function	O
(	O
contractility	O
)	O
and	O
eventually	O
threatens	O
myocyte	B-Cell
viability	O
and	O
organ	B-Organ
function	O
.	O

Ischemic	O
cardiac	B-Tissue
muscle	I-Tissue
is	O
additionally	O
vulnerable	O
because	O
reperfusion	O
is	O
essential	O
for	O
survival	O
but	O
reperfusion	O
itself	O
poses	O
additional	O
stress	O
principally	O
from	O
increased	O
production	O
of	O
free	O
radicals	O
during	O
reoxygenation	O
.	O

The	O
latter	O
effect	O
is	O
called	O
reperfusion	O
injury	O
and	O
can	O
cause	O
as	O
much	O
damage	O
as	O
the	O
ischemia	O
.	O

The	O
treatment	O
possibilities	O
for	O
ischemia	O
-	O
related	O
vascular	B-Multi-tissue_structure
disease	O
are	O
limited	O
.	O

Lipid	B-Simple_chemical
/	O
cholesterol	B-Simple_chemical
-	O
lowering	O
agents	O
,	O
diet	O
and	O
antiplatelet	B-Cell
adherence	O
(	O
aspirin	B-Simple_chemical
)	O
therapy	O
may	O
help	O
slow	O
the	O
progression	O
of	O
vessel	B-Multi-tissue_structure
disease	O
in	O
some	O
instances	O
;	O
but	O
surgical	O
reconstruction	O
may	O
be	O
the	O
only	O
option	O
in	O
advanced	O
stages	O
,	O
and	O
even	O
this	O
is	O
not	O
always	O
an	O
option	O
.	O

An	O
alternative	O
and	O
rather	O
obvious	O
strategy	O
to	O
treat	O
ischemia	O
is	O
to	O
activate	O
endogenous	O
angiogenic	O
or	O
arteriogenic	O
pathways	O
to	O
stimulate	O
revascularization	O
of	O
the	O
tissue	B-Tissue
.	O

The	O
feasibility	O
of	O
such	O
a	O
strategy	O
has	O
now	O
been	O
established	O
through	O
the	O
results	O
of	O
studies	O
over	O
the	O
past	O
decade	O
,	O
and	O
a	O
new	O
discipline	O
called	O
therapeutic	O
angiogenesis	O
has	O
emerged	O
.	O

This	O
review	O
focuses	O
on	O
the	O
application	O
of	O
therapeutic	O
angiogenesis	O
for	O
treating	O
ischemic	O
muscle	B-Organ
disease	O
and	O
includes	O
a	O
critical	O
evaluation	O
of	O
the	O
parameters	O
and	O
limitations	O
of	O
current	O
procedures	O
.	O

The	O
development	O
of	O
this	O
technology	O
has	O
benefited	O
from	O
its	O
application	O
to	O
both	O
peripheral	B-Tissue
and	O
coronary	B-Multi-tissue_structure
artery	I-Multi-tissue_structure
disease	O
and	O
results	O
from	O
both	O
are	O
reviewed	O
here	O
.	O

Expression	O
,	O
regulation	O
,	O
and	O
function	O
of	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1R	I-Gene_or_gene_product
,	O
and	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
binding	I-Gene_or_gene_product
proteins	I-Gene_or_gene_product
in	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
.	O

The	O
vascular	B-Multi-tissue_structure
insulin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
like	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	I-Gene_or_gene_product
IGF	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
system	O
includes	O
the	O
IGFs	B-Gene_or_gene_product
,	O
the	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
(	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1R	I-Gene_or_gene_product
)	O
,	O
and	O
multiple	O
binding	O
proteins	O
.	O

This	O
growth	O
factor	O
system	O
exerts	O
multiple	O
physiologic	O
effects	O
on	O
the	O
vasculature	B-Anatomical_system
through	O
both	O
endocrine	O
and	O
autocrine	O
/	O
paracrine	O
mechanisms	O
.	O

The	O
effects	O
of	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
are	O
mediated	O
principally	O
through	O
the	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1R	I-Gene_or_gene_product
but	O
are	O
modulated	O
by	O
complex	O
interactions	O
with	O
multiple	O
IGF	B-Gene_or_gene_product
binding	I-Gene_or_gene_product
proteins	I-Gene_or_gene_product
that	O
themselves	O
are	O
regulated	O
by	O
phosphorylation	O
,	O
proteolysis	O
,	O
polymerization	O
,	O
and	O
cell	B-Cell
or	O
matrix	B-Cellular_component
association	O
.	O

During	O
the	O
last	O
decade	O
,	O
a	O
significant	O
body	O
of	O
evidence	O
has	O
accumulated	O
,	O
indicating	O
that	O
expression	O
of	O
the	O
components	O
of	O
the	O
IGF	B-Gene_or_gene_product
system	O
are	O
regulated	O
by	O
multiple	O
factors	O
,	O
including	O
growth	O
factors	O
,	O
cytokines	O
,	O
lipoproteins	O
,	O
reactive	B-Simple_chemical
oxygen	I-Simple_chemical
species	I-Simple_chemical
,	O
and	O
hemodynamic	O
forces	O
.	O

In	O
addition	O
,	O
cross	O
-	O
talk	O
between	O
the	O
IGF	B-Gene_or_gene_product
system	O
and	O
other	O
growth	O
factors	O
and	O
integrin	B-Gene_or_gene_product
receptors	O
has	O
been	O
demonstrated	O
.	O

There	O
is	O
accumulating	O
evidence	O
of	O
a	O
role	O
for	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
in	O
multiple	O
vascular	B-Multi-tissue_structure
pathologies	O
,	O
including	O
atherosclerosis	O
,	O
hypertension	O
,	O
restenosis	O
,	O
angiogenesis	O
,	O
and	O
diabetic	O
vascular	B-Multi-tissue_structure
disease	O
.	O

This	O
review	O
will	O
discuss	O
the	O
regulation	O
of	O
expression	O
of	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1R	I-Gene_or_gene_product
,	O
and	O
IGF	B-Gene_or_gene_product
binding	I-Gene_or_gene_product
proteins	I-Gene_or_gene_product
in	O
the	O
vasculature	B-Anatomical_system
and	O
summarize	O
evidence	O
implicating	O
involvement	O
of	O
this	O
system	O
in	O
vascular	B-Multi-tissue_structure
diseases	O
.	O

Microvascular	B-Tissue
density	O
and	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
immunoreactivity	O
as	O
predictors	O
of	O
regional	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
from	O
betel	B-Organism_subdivision
-	O
associated	O
oral	B-Cancer
squamous	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
.	O

PURPOSE	O
:	O
Neovascularization	O
has	O
profound	O
effects	O
on	O
tumor	B-Cancer
growth	O
and	O
metastasis	O
.	O

Vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
is	O
a	O
mitogen	O
that	O
acts	O
exclusively	O
on	O
endothelial	B-Cell
cells	I-Cell
.	O

The	O
roles	O
of	O
miscrovascularity	B-Tissue
density	O
(	O
MVD	O
)	O
and	O
VEGF	B-Gene_or_gene_product
expression	O
in	O
the	O
progression	O
of	O
oral	B-Cancer
squamous	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
(	O
OSCC	B-Cancer
)	O
have	O
been	O
controversial	O
.	O

The	O
purpose	O
of	O
the	O
present	O
study	O
was	O
to	O
measure	O
the	O
MVD	O
and	O
VEGF	B-Gene_or_gene_product
expression	O
in	O
a	O
cohort	O
of	O
patients	B-Organism
with	O
betel	B-Organism_subdivision
-	O
associated	O
OSCC	B-Cancer
and	O
to	O
evaluate	O
for	O
possible	O
clinicopathologic	O
correlations	O
.	O

PATIENTS	O
AND	O
METHODS	O
:	O
The	O
paraffin	B-Cancer
sections	I-Cancer
from	O
49	O
subjects	B-Organism
with	O
OSCC	B-Cancer
were	O
subjected	O
to	O
immunohistochemical	O
studies	O
to	O
measure	O
the	O
highest	O
MVD	O
(	O
h	O
-	O
MVD	O
)	O
and	O
cytoplasmic	B-Organism_substance
immunoreactivity	O
of	O
VEGF	B-Gene_or_gene_product
.	O

The	O
findings	O
in	O
the	O
tissue	B-Cancer
samples	I-Cancer
were	O
analyzed	O
with	O
regard	O
to	O
the	O
patients	B-Organism
'	O
risk	O
factors	O
and	O
clinical	O
course	O
.	O

RESULTS	O
:	O
The	O
OSCC	B-Cancer
samples	I-Cancer
had	O
an	O
average	O
h	O
-	O
MVD	O
score	O
of	O
27	O
.	O
7	O
/	O
mm	O
(	O
2	O
)	O
.	O

VEGF	B-Gene_or_gene_product
immunoreactivity	O
was	O
positive	O
in	O
75	O
.	O
5	O
%	O
of	O
samples	B-Cancer
.	O

Both	O
h	O
-	O
MVD	O
and	O
VEGF	B-Gene_or_gene_product
immunoreactivity	O
were	O
statistically	O
associated	O
with	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
(	O
P	O
=	O
.	O
012	O
and	O
.	O
037	O
,	O
respectively	O
)	O
.	O

A	O
marginally	O
significant	O
association	O
was	O
also	O
noted	O
between	O
the	O
h	O
-	O
MVD	O
and	O
patient	B-Organism
survival	O
(	O
P	O
=	O
.	O
056	O
)	O
.	O

The	O
age	O
and	O
oral	B-Organism_subdivision
habits	O
of	O
patients	B-Organism
,	O
as	O
well	O
as	O
the	O
tumor	B-Cancer
site	O
and	O
size	O
,	O
did	O
not	O
appear	O
to	O
be	O
correlated	O
with	O
h	O
-	O
MVD	O
or	O
VEGF	B-Gene_or_gene_product
immunoreactivity	O
.	O

CONCLUSION	O
:	O
The	O
data	O
suggest	O
that	O
both	O
h	O
-	O
MVD	O
and	O
VEGF	B-Gene_or_gene_product
immunoreactivity	O
may	O
be	O
useful	O
predictors	O
for	O
the	O
progression	O
of	O
a	O
subset	O
of	O
OSCC	B-Cancer
associated	O
mostly	O
with	O
betel	B-Organism_subdivision
use	O
.	O

Antiangiogenesis	O
therapy	O
might	O
have	O
a	O
role	O
in	O
reducing	O
regional	O
metastasis	O
.	O

Promotion	O
of	O
tumorigenesis	O
by	O
heterozygous	O
disruption	O
of	O
the	O
beclin	B-Gene_or_gene_product
1	I-Gene_or_gene_product
autophagy	O
gene	O
.	O

Malignant	B-Cell
cells	I-Cell
often	O
display	O
defects	O
in	O
autophagy	O
,	O
an	O
evolutionarily	O
conserved	O
pathway	O
for	O
degrading	O
long	O
-	O
lived	O
proteins	O
and	O
cytoplasmic	B-Cellular_component
organelles	I-Cellular_component
.	O

However	O
,	O
as	O
yet	O
,	O
there	O
is	O
no	O
genetic	O
evidence	O
for	O
a	O
role	O
of	O
autophagy	O
genes	O
in	O
tumor	B-Cancer
suppression	O
.	O

The	O
beclin	B-Gene_or_gene_product
1	I-Gene_or_gene_product
autophagy	O
gene	O
is	O
monoallelically	O
deleted	O
in	O
40	O
-	O
75	O
%	O
of	O
cases	O
of	O
human	B-Organism
sporadic	O
breast	B-Cancer
,	O
ovarian	B-Cancer
,	O
and	O
prostate	B-Cancer
cancer	I-Cancer
.	O

Therefore	O
,	O
we	O
used	O
a	O
targeted	O
mutant	O
mouse	B-Organism
model	O
to	O
test	O
the	O
hypothesis	O
that	O
monoallelic	O
deletion	O
of	O
beclin	B-Gene_or_gene_product
1	I-Gene_or_gene_product
promotes	O
tumorigenesis	O
.	O

Here	O
we	O
show	O
that	O
heterozygous	O
disruption	O
of	O
beclin	B-Gene_or_gene_product
1	I-Gene_or_gene_product
increases	O
the	O
frequency	O
of	O
spontaneous	O
malignancies	B-Cancer
and	O
accelerates	O
the	O
development	O
of	O
hepatitis	B-Organism
B	I-Organism
virus	I-Organism
-	O
induced	O
premalignant	B-Pathological_formation
lesions	I-Pathological_formation
.	O

Molecular	O
analyses	O
of	O
tumors	B-Cancer
in	O
beclin	B-Gene_or_gene_product
1	I-Gene_or_gene_product
heterozygous	O
mice	B-Organism
show	O
that	O
the	O
remaining	O
wild	O
-	O
type	O
allele	O
is	O
neither	O
mutated	O
nor	O
silenced	O
.	O

Furthermore	O
,	O
beclin	B-Gene_or_gene_product
1	I-Gene_or_gene_product
heterozygous	O
disruption	O
results	O
in	O
increased	O
cellular	B-Cell
proliferation	O
and	O
reduced	O
autophagy	O
in	O
vivo	O
.	O

These	O
findings	O
demonstrate	O
that	O
beclin	B-Gene_or_gene_product
1	I-Gene_or_gene_product
is	O
a	O
haplo	O
-	O
insufficient	O
tumor	B-Cancer
-	O
suppressor	O
gene	O
and	O
provide	O
genetic	O
evidence	O
that	O
autophagy	O
is	O
a	O
novel	O
mechanism	O
of	O
cell	B-Cell
-	O
growth	O
control	O
and	O
tumor	B-Cancer
suppression	O
.	O

Thus	O
,	O
mutation	O
of	O
beclin	B-Gene_or_gene_product
1	I-Gene_or_gene_product
or	O
other	O
autophagy	O
genes	O
may	O
contribute	O
to	O
the	O
pathogenesis	O
of	O
human	B-Organism
cancers	B-Cancer
.	O

Effects	O
of	O
dietary	O
folate	B-Simple_chemical
on	O
ulcerative	B-Pathological_formation
colitis	I-Pathological_formation
-	O
associated	O
colorectal	B-Multi-tissue_structure
carcinogenesis	O
in	O
the	O
interleukin	B-Gene_or_gene_product
2	I-Gene_or_gene_product
-	O
and	O
beta	B-Gene_or_gene_product
(	I-Gene_or_gene_product
2	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	I-Gene_or_gene_product
microglobulin	I-Gene_or_gene_product
-	O
deficient	O
mice	B-Organism
.	O

Folate	B-Simple_chemical
supplementation	O
may	O
reduce	O
the	O
risk	O
of	O
colorectal	B-Pathological_formation
dysplasia	I-Pathological_formation
and	O
cancer	B-Cancer
in	O
subjects	O
with	O
chronic	O
ulcerative	B-Pathological_formation
colitis	I-Pathological_formation
(	O
UC	B-Pathological_formation
)	O
.	O

The	O
interleukin	B-Gene_or_gene_product
(	I-Gene_or_gene_product
IL	I-Gene_or_gene_product
)	I-Gene_or_gene_product
2	I-Gene_or_gene_product
-	O
and	O
beta	B-Gene_or_gene_product
(	I-Gene_or_gene_product
2	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	I-Gene_or_gene_product
microglobulin	I-Gene_or_gene_product
(	O
beta	B-Gene_or_gene_product
(	I-Gene_or_gene_product
2	I-Gene_or_gene_product
)	I-Gene_or_gene_product
m	I-Gene_or_gene_product
)	O
-	O
deficient	O
(	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
(	O
null	O
)	O
x	O
beta	B-Gene_or_gene_product
(	I-Gene_or_gene_product
2	I-Gene_or_gene_product
)	I-Gene_or_gene_product
m	I-Gene_or_gene_product
(	O
null	O
)	O
)	O
mice	B-Organism
spontaneously	O
develop	O
colon	B-Cancer
cancer	I-Cancer
in	O
the	O
setting	O
of	O
chronic	O
UC	B-Pathological_formation
.	O

This	O
study	O
investigated	O
the	O
effects	O
of	O
dietary	O
folate	B-Simple_chemical
on	O
the	O
development	O
of	O
UC	B-Pathological_formation
-	O
associated	O
colon	B-Cancer
cancer	I-Cancer
in	O
the	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
(	O
null	O
)	O
x	O
beta	B-Gene_or_gene_product
(	I-Gene_or_gene_product
2	I-Gene_or_gene_product
)	I-Gene_or_gene_product
m	I-Gene_or_gene_product
(	O
null	O
)	O
mice	B-Organism
.	O

Weaning	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
(	O
null	O
)	O
x	O
beta	B-Gene_or_gene_product
(	I-Gene_or_gene_product
2	I-Gene_or_gene_product
)	I-Gene_or_gene_product
m	I-Gene_or_gene_product
(	O
null	O
)	O
mice	B-Organism
were	O
randomized	O
to	O
receive	O
0	O
(	O
deficient	O
;	O
n	O
=	O
40	O
)	O
,	O
2	O
(	O
basal	O
requirement	O
;	O
control	O
;	O
n	O
=	O
46	O
)	O
,	O
or	O
8	O
(	O
supplemented	O
;	O
n	O
=	O
36	O
)	O
mg	O
folate	B-Simple_chemical
/	O
kg	O
diet	O
for	O
32	O
weeks	O
.	O

At	O
necropsy	O
,	O
all	O
macroscopic	O
colonic	B-Cancer
tumors	I-Cancer
were	O
identified	O
and	O
histologically	O
classified	O
as	O
dysplasia	B-Pathological_formation
or	O
adenocarcinoma	B-Cancer
.	O

The	O
incidence	O
of	O
high	B-Pathological_formation
-	I-Pathological_formation
grade	I-Pathological_formation
lesions	I-Pathological_formation
(	O
high	B-Pathological_formation
-	I-Pathological_formation
grade	I-Pathological_formation
dysplasia	I-Pathological_formation
/	O
carcinoma	B-Cancer
in	O
situ	O
and	O
invasive	O
adenocarcinoma	B-Cancer
)	O
in	O
the	O
folate	B-Simple_chemical
-	O
supplemented	O
group	O
was	O
46	O
%	O
lower	O
than	O
that	O
in	O
the	O
control	O
group	O
(	O
35	O
.	O
3	O
%	O
versus	O
65	O
.	O
1	O
%	O
,	O
P	O
=	O
0	O
.	O
009	O
)	O
.	O

The	O
incidence	O
of	O
high	B-Pathological_formation
-	I-Pathological_formation
grade	I-Pathological_formation
lesions	I-Pathological_formation
in	O
the	O
folate	B-Simple_chemical
-	O
deficient	O
group	O
was	O
also	O
49	O
%	O
lower	O
than	O
that	O
in	O
the	O
control	O
group	O
(	O
33	O
.	O
3	O
%	O
versus	O
65	O
.	O
1	O
%	O
,	O
P	O
=	O
0	O
.	O
007	O
)	O
.	O

The	O
higher	O
mortality	O
rate	O
in	O
the	O
folate	B-Simple_chemical
-	O
deficient	O
group	O
compared	O
with	O
the	O
other	O
two	O
groups	O
(	O
25	O
%	O
versus	O
6	O
.	O
5	O
%	O
and	O
5	O
.	O
6	O
%	O
,	O
P	O
<	O
0	O
.	O
02	O
)	O
partially	O
accounted	O
for	O
the	O
low	O
incidence	O
of	O
high	B-Pathological_formation
-	I-Pathological_formation
grade	I-Pathological_formation
lesions	I-Pathological_formation
in	O
this	O
group	O
.	O

These	O
data	O
indicate	O
that	O
dietary	O
folate	B-Simple_chemical
supplementation	O
at	O
4x	O
the	O
basal	O
dietary	O
requirement	O
significantly	O
suppresses	O
UC	B-Pathological_formation
-	O
associated	O
colorectal	B-Multi-tissue_structure
carcinogenesis	O
in	O
the	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
(	O
null	O
)	O
x	O
beta	B-Gene_or_gene_product
(	I-Gene_or_gene_product
2	I-Gene_or_gene_product
)	I-Gene_or_gene_product
m	I-Gene_or_gene_product
(	O
null	O
)	O
mice	B-Organism
.	O

These	O
data	O
also	O
suggest	O
that	O
folate	B-Simple_chemical
deficiency	O
may	O
inhibit	O
colorectal	B-Multi-tissue_structure
carcinogenesis	O
in	O
chronic	O
UC	B-Pathological_formation
.	O

However	O
,	O
the	O
high	O
mortality	O
observed	O
in	O
the	O
folate	B-Simple_chemical
-	O
deficient	O
group	O
precludes	O
a	O
definitive	O
conclusion	O
concerning	O
the	O
effect	O
of	O
folate	B-Simple_chemical
deficiency	O
on	O
UC	B-Pathological_formation
-	O
associated	O
colorectal	B-Multi-tissue_structure
carcinogenesis	O
in	O
this	O
model	O
.	O

Effect	O
of	O
antitumor	B-Cancer
agents	O
on	O
cytotoxicity	O
induction	O
by	O
sodium	B-Simple_chemical
fluoride	I-Simple_chemical
.	O

We	O
have	O
recently	O
found	O
that	O
sodium	B-Simple_chemical
fluoride	I-Simple_chemical
(	O
NaF	B-Simple_chemical
)	O
induced	O
apoptotic	O
cell	B-Cell
death	O
in	O
tumor	B-Cell
cell	I-Cell
lines	I-Cell
.	O

We	O
investigated	O
here	O
whether	O
6	O
popular	O
antitumor	B-Cancer
compounds	O
modify	O
the	O
cytotoxic	O
activity	O
of	O
NaF	B-Simple_chemical
against	O
human	B-Organism
squamous	B-Cancer
cell	I-Cancer
carcinoma	I-Cancer
(	O
HSC	B-Cell
-	I-Cell
2	I-Cell
)	I-Cell
and	O
human	B-Organism
promyelocytic	B-Cancer
leukemia	I-Cancer
(	O
HL	B-Cell
-	I-Cell
60	I-Cell
)	I-Cell
cell	I-Cell
lines	I-Cell
.	O

Cytotoxic	O
concentrations	O
of	O
cisplatin	B-Simple_chemical
,	O
etoposide	B-Simple_chemical
,	O
doxorubicin	B-Simple_chemical
or	O
peplomycin	B-Simple_chemical
(	O
tentatively	O
termed	O
as	O
Group	O
I	O
compounds	O
)	O
,	O
but	O
not	O
methotrexate	B-Simple_chemical
and	O
5	B-Simple_chemical
-	I-Simple_chemical
FU	I-Simple_chemical
(	O
tentatively	O
termed	O
as	O
Group	O
II	O
compounds	O
)	O
,	O
enhanced	O
the	O
cytotoxic	O
activity	O
of	O
NaF	B-Simple_chemical
.	O

NaF	B-Simple_chemical
and	O
Group	O
I	O
compounds	O
induced	O
internucleosomal	O
DNA	B-Cellular_component
fragmentation	O
in	O
HL	B-Cell
-	I-Cell
60	I-Cell
cells	I-Cell
,	O
whereas	O
Group	O
II	O
compounds	O
were	O
inactive	O
even	O
in	O
the	O
presence	O
of	O
NaF	B-Simple_chemical
.	O

Most	O
Group	O
I	O
compounds	O
except	O
doxorubicin	B-Simple_chemical
(	O
which	O
induced	O
DNA	B-Cellular_component
fragmentation	O
less	O
effectively	O
than	O
others	O
)	O
activated	O
caspase	B-Gene_or_gene_product
3	I-Gene_or_gene_product
more	O
efficiently	O
than	O
Group	O
II	O
compounds	O
.	O

Caspase	B-Gene_or_gene_product
8	I-Gene_or_gene_product
(	O
involved	O
in	O
non	O
-	O
mitochondrial	B-Cellular_component
extrinsic	O
pathway	O
)	O
and	O
caspase	B-Gene_or_gene_product
9	I-Gene_or_gene_product
(	O
involved	O
in	O
mitochondrial	B-Cellular_component
intrinsic	O
pathway	O
)	O
were	O
also	O
activated	O
,	O
but	O
to	O
a	O
much	O
lesser	O
extent	O
.	O

NaF	B-Simple_chemical
reduced	O
the	O
glucose	B-Simple_chemical
consumption	O
at	O
early	O
stage	O
,	O
possibly	O
by	O
inhibition	O
of	O
glycolysis	O
,	O
whereas	O
cisplatin	B-Simple_chemical
and	O
etoposide	B-Simple_chemical
reduced	O
the	O
glucose	B-Simple_chemical
consumption	O
at	O
later	O
stage	O
,	O
suggesting	O
that	O
early	O
decline	O
of	O
glucose	B-Simple_chemical
consumption	O
is	O
rather	O
specific	O
to	O
NaF	B-Simple_chemical
.	O

Morphogenesis	O
of	O
embryonic	B-Multi-tissue_structure
CNS	I-Multi-tissue_structure
vessels	I-Multi-tissue_structure
.	O

This	O
chapter	O
focuses	O
on	O
the	O
morphology	O
of	O
blood	B-Multi-tissue_structure
vessel	I-Multi-tissue_structure
formation	O
in	O
and	O
around	O
the	O
early	O
central	B-Anatomical_system
nervous	I-Anatomical_system
system	I-Anatomical_system
(	O
CNS	B-Anatomical_system
,	O
i	O
.	O
e	O
.	O
,	O
brain	B-Organ
and	O
spinal	B-Organ
cord	I-Organ
)	O
of	O
avian	B-Organism
embryos	B-Developing_anatomical_structure
.	O

We	O
discuss	O
cell	B-Cell
lineages	O
,	O
proliferation	O
and	O
interactions	O
of	O
endothelial	B-Cell
cells	I-Cell
,	O
pericytes	B-Cell
and	O
smooth	B-Cell
muscle	I-Cell
cells	I-Cell
,	O
and	O
macrophages	B-Cell
.	O

Due	O
to	O
space	O
limitations	O
,	O
we	O
can	O
not	O
review	O
the	O
molecular	O
control	O
of	O
CNS	B-Anatomical_system
angiogenesis	O
,	O
but	O
refer	O
the	O
reader	O
to	O
other	O
chapters	O
in	O
this	O
book	O
and	O
to	O
recent	O
publications	O
on	O
the	O
assembly	O
of	O
the	O
vasculature	B-Multi-tissue_structure
(	O
1	O
,	O
2	O
)	O
.	O

The	O
candidate	O
tumour	B-Cancer
suppressor	O
protein	O
ING4	B-Gene_or_gene_product
regulates	O
brain	B-Cancer
tumour	I-Cancer
growth	O
and	O
angiogenesis	O
.	O

Gliomas	B-Cancer
are	O
the	O
most	O
common	O
primary	O
tumours	B-Cancer
of	O
the	O
central	B-Anatomical_system
nervous	I-Anatomical_system
system	I-Anatomical_system
,	O
with	O
nearly	O
15	O
,	O
000	O
diagnosed	O
annually	O
in	O
the	O
United	O
States	O
and	O
a	O
lethality	O
approaching	O
80	O
%	O
within	O
the	O
first	O
year	O
of	O
glioblastoma	B-Cancer
diagnosis	O
.	O

The	O
marked	O
induction	O
of	O
angiogenesis	O
in	O
glioblastomas	B-Cancer
suggests	O
that	O
it	O
is	O
a	O
necessary	O
part	O
of	O
malignant	O
progression	O
;	O
however	O
,	O
the	O
precise	O
molecular	O
mechanisms	O
underlying	O
the	O
regulation	O
of	O
brain	B-Cancer
tumour	I-Cancer
growth	O
and	O
angiogenesis	O
remain	O
unresolved	O
.	O

Here	O
we	O
report	O
that	O
a	O
candidate	O
tumour	B-Cancer
suppressor	O
gene	O
,	O
ING4	B-Gene_or_gene_product
,	O
is	O
involved	O
in	O
regulating	O
brain	B-Cancer
tumour	I-Cancer
growth	O
and	O
angiogenesis	O
.	O

Expression	O
of	O
ING4	B-Gene_or_gene_product
is	O
significantly	O
reduced	O
in	O
gliomas	B-Cancer
as	O
compared	O
with	O
normal	O
human	B-Organism
brain	B-Tissue
tissue	I-Tissue
,	O
and	O
the	O
extent	O
of	O
reduction	O
correlates	O
with	O
the	O
progression	O
from	O
lower	O
to	O
higher	O
grades	O
of	O
tumours	B-Cancer
.	O

In	O
mice	B-Organism
,	O
xenografts	B-Cancer
of	O
human	B-Organism
glioblastoma	B-Cancer
U87MG	I-Cancer
,	O
which	O
has	O
decreased	O
expression	O
of	O
ING4	B-Gene_or_gene_product
,	O
grow	O
significantly	O
faster	O
and	O
have	O
higher	O
vascular	B-Multi-tissue_structure
volume	O
fractions	O
than	O
control	O
tumours	B-Cancer
.	O

We	O
show	O
that	O
ING4	B-Gene_or_gene_product
physically	O
interacts	O
with	O
p65	B-Gene_or_gene_product
(	O
RelA	B-Gene_or_gene_product
)	O
subunit	O
of	O
nuclear	O
factor	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
,	O
and	O
that	O
ING4	B-Gene_or_gene_product
regulates	O
brain	B-Cancer
tumour	I-Cancer
angiogenesis	O
through	O
transcriptional	O
repression	O
of	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
-	O
responsive	O
genes	O
.	O

These	O
results	O
indicate	O
that	O
ING4	B-Gene_or_gene_product
has	O
an	O
important	O
role	O
in	O
brain	B-Cancer
tumour	I-Cancer
pathogenesis	O
.	O

TGFbeta1	B-Gene_or_gene_product
,	O
back	O
to	O
the	O
future	O
:	O
revisiting	O
its	O
role	O
as	O
a	O
transforming	O
growth	O
factor	O
.	O

TGFbeta1	B-Gene_or_gene_product
was	O
initially	O
identified	O
in	O
culture	O
media	O
from	O
transformed	B-Cell
cells	I-Cell
as	O
part	O
of	O
a	O
factor	O
that	O
could	O
produce	O
a	O
transformed	O
phenotype	O
in	O
a	O
nontransformed	B-Cell
cell	I-Cell
line	O
.	O

Subsequently	O
this	O
activity	O
was	O
separated	O
into	O
TGFbeta	B-Gene_or_gene_product
and	O
TGFalpha	B-Gene_or_gene_product
an	O
EGF	B-Gene_or_gene_product
receptor	O
ligand	O
.	O

With	O
the	O
discovery	O
that	O
TGFbeta1	B-Gene_or_gene_product
was	O
a	O
potent	O
growth	O
inhibitor	O
of	O
epithelial	B-Cell
cells	I-Cell
,	O
and	O
the	O
identification	O
of	O
inactivating	O
mutations	O
within	O
the	O
TGFbeta1	B-Gene_or_gene_product
signaling	O
pathway	O
in	O
cancers	B-Cancer
it	O
became	O
clear	O
that	O
TGFbeta1	B-Gene_or_gene_product
signaling	O
is	O
a	O
tumor	B-Cancer
suppressor	O
pathway	O
for	O
early	O
stages	O
of	O
cancer	B-Cancer
.	O

However	O
many	O
human	B-Organism
carcinomas	B-Cancer
overexpress	O
TGFbeta1	B-Gene_or_gene_product
and	O
this	O
is	O
associated	O
with	O
poor	O
patient	B-Organism
prognosis	O
and	O
increased	O
frequency	O
of	O
metastasis	O
.	O

Similar	O
results	O
have	O
been	O
obtained	O
with	O
tumor	B-Cell
cell	I-Cell
lines	I-Cell
and	O
experimental	O
animal	O
models	O
.	O

Thus	O
stage	O
specific	O
duality	O
of	O
function	O
is	O
the	O
emerging	O
paradigm	O
for	O
the	O
role	O
of	O
TGFbeta1	B-Gene_or_gene_product
in	O
cancer	B-Cancer
.	O

This	O
review	O
will	O
focus	O
on	O
the	O
evidence	O
for	O
TGFbeta1	B-Gene_or_gene_product
as	O
a	O
tumor	B-Cancer
promoting	O
and	O
metastasis	O
factor	O
and	O
examine	O
the	O
biological	O
and	O
molecular	O
basis	O
for	O
these	O
effects	O
.	O

It	O
is	O
proposed	O
that	O
the	O
switch	O
from	O
tumor	B-Cancer
suppressor	O
to	O
oncogene	O
reflects	O
genetic	O
or	O
epigenetic	O
alterations	O
in	O
signaling	O
pathways	O
in	O
tumor	B-Cell
cells	I-Cell
that	O
alter	O
the	O
readout	O
from	O
the	O
TGFbeta1	B-Gene_or_gene_product
pathway	O
.	O

Role	O
of	O
thrombin	B-Gene_or_gene_product
in	O
angiogenesis	O
and	O
tumor	B-Cancer
progression	O
.	O

Clinical	O
,	O
laboratory	O
,	O
histopathological	O
,	O
and	O
pharmacological	O
evidence	O
support	O
the	O
notion	O
that	O
the	O
coagulation	O
system	O
,	O
which	O
is	O
activated	O
in	O
most	O
cancer	B-Cancer
patients	B-Organism
,	O
plays	O
an	O
important	O
role	O
in	O
tumor	B-Cancer
biology	O
.	O

Our	O
laboratory	O
has	O
provided	O
evidence	O
that	O
thrombin	B-Gene_or_gene_product
activates	O
angiogenesis	O
,	O
a	O
process	O
which	O
is	O
essential	O
in	O
tumor	B-Cancer
growth	O
and	O
metastasis	O
.	O

This	O
event	O
is	O
independent	O
of	O
fibrin	B-Gene_or_gene_product
formation	O
.	O

At	O
the	O
cellular	B-Cell
level	O
many	O
actions	O
of	O
thrombin	B-Gene_or_gene_product
can	O
contribute	O
to	O
activation	O
of	O
angiogenesis	O
:	O
(	O
1	O
)	O
.	O

Thrombin	B-Gene_or_gene_product
decreases	O
the	O
ability	O
of	O
endothelial	B-Cell
cells	I-Cell
to	O
attach	O
to	O
basement	B-Cellular_component
membrane	I-Cellular_component
proteins	O
.	O

(	O
2	O
)	O
.	O

Thrombin	B-Gene_or_gene_product
greatly	O
potentiates	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	O
(	O
VEGF	B-Gene_or_gene_product
-	O
)	O
induced	O
endothelial	B-Cell
cell	I-Cell
proliferation	O
.	O

This	O
potentiation	O
is	O
accompanied	O
by	O
up	O
-	O
regulation	O
of	O
the	O
expression	O
of	O
VEGF	B-Gene_or_gene_product
receptors	O
(	O
kinase	B-Gene_or_gene_product
insert	I-Gene_or_gene_product
domain	I-Gene_or_gene_product
-	I-Gene_or_gene_product
containing	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
[	O
KDR	B-Gene_or_gene_product
]	O
and	O
fms	B-Gene_or_gene_product
-	I-Gene_or_gene_product
like	I-Gene_or_gene_product
tyrosine	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
[	O
Flt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
]	O
)	O
.	O

(	O
3	O
)	O
.	O

Thrombin	B-Gene_or_gene_product
increases	O
the	O
mRNA	O
and	O
protein	O
levels	O
of	O
alpha	B-Gene_or_gene_product
(	I-Gene_or_gene_product
v	I-Gene_or_gene_product
)	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
(	I-Gene_or_gene_product
3	I-Gene_or_gene_product
)	I-Gene_or_gene_product
integrin	I-Gene_or_gene_product
and	O
serves	O
as	O
a	O
ligand	O
to	O
this	O
receptor	O
.	O

Furthermore	O
,	O
thrombin	B-Gene_or_gene_product
increases	O
the	O
secretion	O
of	O
VEGF	B-Gene_or_gene_product
and	O
enhances	O
the	O
expression	O
and	O
protein	O
synthesis	O
of	O
matrix	B-Gene_or_gene_product
metalloprotease	I-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
and	O
alpha	B-Gene_or_gene_product
(	I-Gene_or_gene_product
v	I-Gene_or_gene_product
)	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
(	I-Gene_or_gene_product
3	I-Gene_or_gene_product
)	I-Gene_or_gene_product
integrin	I-Gene_or_gene_product
in	O
human	B-Organism
prostate	B-Cell
cancer	I-Cell
PC	I-Cell
-	I-Cell
3	I-Cell
cells	I-Cell
.	O

These	O
results	O
could	O
explain	O
the	O
angiogenic	O
and	O
tumor	B-Cancer
-	O
promoting	O
effect	O
of	O
thrombin	B-Gene_or_gene_product
and	O
provide	O
the	O
basis	O
for	O
development	O
of	O
thrombin	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
mimetics	O
or	O
antagonists	O
for	O
therapeutic	O
application	O
.	O

NIH3T3	B-Cell
transfectant	I-Cell
containing	O
human	B-Organism
K	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
oncogene	O
shows	O
enhanced	O
metastatic	O
activity	O
after	O
in	O
vivo	O
tumor	B-Cancer
growth	O
or	O
co	O
-	O
culture	O
with	O
fibroblasts	B-Cell
.	O

A	O
clone	B-Cell
of	O
NIH3T3	B-Cell
transformant	I-Cell
(	O
H	B-Cell
-	I-Cell
3	I-Cell
)	O
,	O
obtained	O
by	O
transfecting	O
genomic	O
DNA	B-Cellular_component
of	O
a	O
human	B-Organism
colon	B-Cell
carcinoma	I-Cell
cell	I-Cell
line	I-Cell
,	O
contains	O
human	B-Organism
K	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
oncogene	O
and	O
yields	O
metastatic	O
pulmonary	B-Pathological_formation
nodules	I-Pathological_formation
after	O
intravenous	B-Immaterial_anatomical_entity
injection	O
of	O
the	O
cells	B-Cell
into	O
nude	B-Organism
mice	I-Organism
.	O

This	O
metastatic	O
ability	O
was	O
enhanced	O
remarkably	O
after	O
in	O
vivo	O
tumor	B-Cancer
growth	O
(	O
subcutaneous	O
tumor	B-Cancer
formation	O
in	O
nude	B-Organism
mice	I-Organism
)	O
accompanied	O
by	O
increased	O
mRNA	O
expression	O
and	O
gene	O
amplification	O
of	O
the	O
human	B-Organism
-	O
derived	O
K	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
oncogene	O
,	O
while	O
it	O
declined	O
gradually	O
as	O
the	O
passage	O
number	O
increased	O
in	O
vitro	O
,	O
with	O
corresponding	O
decreases	O
of	O
gene	O
amplification	O
and	O
mRNA	O
expression	O
.	O

Six	O
subclones	B-Cell
were	O
randomly	O
selected	O
from	O
H	B-Cell
-	I-Cell
3	I-Cell
cells	I-Cell
which	O
had	O
been	O
subcultured	O
to	O
passage	O
22	O
.	O

All	O
of	O
the	O
clones	B-Cell
in	O
culture	O
showed	O
almost	O
the	O
same	O
low	O
level	O
of	O
metastatic	O
ability	O
and	O
exhibited	O
little	O
K	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
oncogene	O
amplification	O
with	O
correspondingly	O
low	O
mRNA	O
expression	O
.	O

However	O
,	O
after	O
they	O
formed	O
tumors	B-Cancer
in	O
nude	B-Organism
mice	I-Organism
,	O
every	O
clone	B-Cell
acquired	O
high	O
metastatic	O
ability	O
and	O
the	O
gene	O
amplification	O
increased	O
,	O
with	O
elevated	O
mRNA	O
expression	O
.	O

These	O
experimental	O
facts	O
indicated	O
that	O
acquisition	O
of	O
metastatic	O
ability	O
coupled	O
with	O
the	O
function	O
of	O
K	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
oncogene	O
was	O
conditional	O
in	O
nature	O
,	O
being	O
strongly	O
affected	O
by	O
in	O
vivo	O
tumor	B-Cancer
circumstances	O
.	O

The	O
low	O
metastatic	O
and	O
G	B-Simple_chemical
-	I-Simple_chemical
418	I-Simple_chemical
-	O
resistant	O
H	B-Cell
-	I-Cell
3	I-Cell
cells	I-Cell
were	O
co	O
-	O
cultured	O
with	O
BALB	B-Cell
/	I-Cell
c3T3	I-Cell
fibroblasts	I-Cell
for	O
2	O
-	O
4	O
weeks	O
.	O

After	O
removal	O
of	O
fibroblasts	B-Cell
by	O
exposure	O
to	O
G	B-Simple_chemical
-	I-Simple_chemical
418	I-Simple_chemical
,	O
the	O
tumor	B-Cell
cells	I-Cell
exhibited	O
increased	O
metastatic	O
ability	O
and	O
human	B-Organism
K	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
oncogene	O
mRNA	O
,	O
suggesting	O
an	O
intimate	O
interaction	O
between	O
H	B-Cell
-	I-Cell
3	I-Cell
cells	I-Cell
and	O
fibroblasts	B-Cell
influencing	O
the	O
function	O
of	O
transfected	O
human	B-Organism
K	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
oncogene	O
.	O

Fibroblasts	B-Cell
of	O
the	O
host	O
animal	B-Organism
may	O
thus	O
have	O
an	O
important	O
role	O
in	O
generating	O
enhanced	O
metastatic	O
activity	O
of	O
H	B-Cell
-	I-Cell
3	I-Cell
cells	I-Cell
.	O

Lactoferrin	B-Gene_or_gene_product
enhances	O
Fas	B-Gene_or_gene_product
expression	O
and	O
apoptosis	O
in	O
the	O
colon	B-Multi-tissue_structure
mucosa	I-Multi-tissue_structure
of	O
azoxymethane	B-Simple_chemical
-	O
treated	O
rats	B-Organism
.	O

Bovine	B-Organism
lactoferrin	B-Gene_or_gene_product
,	O
a	O
multifunctional	O
glycoprotein	O
,	O
has	O
been	O
shown	O
to	O
strongly	O
inhibit	O
development	O
of	O
azoxymethane	B-Simple_chemical
(	O
AOM	B-Simple_chemical
)	O
-	O
induced	O
rat	B-Organism
colon	B-Cancer
tumors	I-Cancer
.	O

Little	O
,	O
however	O
,	O
is	O
known	O
about	O
the	O
inhibitory	O
mechanisms	O
.	O

We	O
have	O
demonstrated	O
recently	O
that	O
lactoferrin	B-Gene_or_gene_product
enhances	O
the	O
expression	O
of	O
a	O
member	O
of	O
the	O
tumor	B-Gene_or_gene_product
necrosis	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
family	O
,	O
Fas	B-Gene_or_gene_product
,	O
in	O
the	O
colon	B-Multi-tissue_structure
mucosa	I-Multi-tissue_structure
during	O
both	O
early	O
and	O
late	O
stages	O
of	O
carcinogenesis	O
.	O

Thus	O
,	O
Fas	B-Gene_or_gene_product
could	O
be	O
involved	O
in	O
bovine	B-Organism
lactoferrin	B-Gene_or_gene_product
-	O
mediated	O
inhibition	O
of	O
tumor	B-Cancer
development	O
.	O

To	O
investigate	O
this	O
possibility	O
,	O
we	O
studied	O
the	O
influence	O
of	O
bovine	B-Organism
lactoferrin	B-Gene_or_gene_product
on	O
Fas	B-Gene_or_gene_product
-	O
mediated	O
apoptosis	O
with	O
regard	O
to	O
expression	O
of	O
Fas	B-Gene_or_gene_product
,	O
activation	O
of	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
and	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
,	O
and	O
DNA	B-Cellular_component
fragmentation	O
in	O
the	O
colon	B-Multi-tissue_structure
mucosa	I-Multi-tissue_structure
of	O
AOM	B-Simple_chemical
-	O
treated	O
rats	B-Organism
.	O

Western	O
blot	O
analysis	O
demonstrated	O
a	O
>	O
2	O
.	O
5	O
-	O
fold	O
increase	O
in	O
Fas	B-Gene_or_gene_product
protein	O
expression	O
,	O
as	O
well	O
as	O
elevation	O
of	O
the	O
active	O
forms	O
of	O
both	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
and	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
.	O

Immunohistochemical	O
analysis	O
revealed	O
Fas	B-Gene_or_gene_product
-	O
positive	O
cells	O
and	O
apoptotic	B-Cell
cells	I-Cell
preferentially	O
within	O
the	O
proximal	O
colon	B-Organ
region	O
,	O
clearly	O
at	O
the	O
site	O
of	O
bovine	B-Organism
lactoferrin	B-Gene_or_gene_product
-	O
mediated	O
tumor	B-Cancer
inhibition	O
.	O

These	O
results	O
suggest	O
that	O
apoptosis	O
caused	O
by	O
elevated	O
expression	O
of	O
Fas	B-Gene_or_gene_product
is	O
involved	O
in	O
chemoprevention	O
by	O
lactoferrin	B-Gene_or_gene_product
of	O
colon	B-Organ
carcinogenesis	O
.	O

Dynein	B-Gene_or_gene_product
light	I-Gene_or_gene_product
chain	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
a	O
p21	B-Gene_or_gene_product
-	I-Gene_or_gene_product
activated	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
interacting	O
substrate	O
,	O
promotes	O
cancerous	B-Cancer
phenotypes	O
.	O

We	O
identified	O
dynein	B-Gene_or_gene_product
light	I-Gene_or_gene_product
chain	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
DLC1	B-Gene_or_gene_product
)	O
as	O
a	O
physiologic	O
substrate	O
of	O
p21	B-Gene_or_gene_product
-	I-Gene_or_gene_product
activated	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
Pak1	B-Gene_or_gene_product
)	O
.	O

Pak1	B-Gene_or_gene_product
-	O
DLC1	B-Gene_or_gene_product
interaction	O
plays	O
an	O
essential	O
role	O
in	O
cell	B-Cell
survival	O
,	O
which	O
depends	O
on	O
Pak1	B-Gene_or_gene_product
'	O
s	O
phosphorylation	O
of	O
DLC1	B-Gene_or_gene_product
on	O
Ser88	B-Amino_acid
.	O

Pak1	B-Gene_or_gene_product
associates	O
with	O
the	O
complex	O
of	O
DLC1	B-Gene_or_gene_product
and	O
BimL	B-Gene_or_gene_product
,	O
a	O
proapoptotic	O
BH3	O
-	O
only	O
protein	O
,	O
and	O
phosphorylates	O
both	O
proteins	O
.	O

Phosphorylation	O
of	O
BimL	B-Gene_or_gene_product
by	O
Pak1	B-Gene_or_gene_product
prevents	O
it	O
from	O
interacting	O
with	O
and	O
inactivation	O
of	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
an	O
antiapoptotic	O
protein	O
.	O

Overexpression	O
of	O
DLC1	B-Gene_or_gene_product
but	O
not	O
DLC1	B-Gene_or_gene_product
-	O
Ser88Ala	B-Amino_acid
mutant	O
promotes	O
cancerous	B-Cancer
properties	O
of	O
breast	B-Cell
cancer	I-Cell
cells	I-Cell
.	O

DLC1	B-Gene_or_gene_product
protein	O
level	O
is	O
elevated	O
in	O
more	O
than	O
90	O
%	O
of	O
human	B-Organism
breast	B-Cancer
tumors	I-Cancer
.	O

The	O
regulation	O
of	O
cell	B-Cell
survival	O
functions	O
by	O
Pak1	B-Gene_or_gene_product
-	O
DLC1	B-Gene_or_gene_product
interaction	O
represents	O
a	O
novel	O
mechanism	O
by	O
which	O
a	O
signaling	O
kinase	O
might	O
regulate	O
the	O
cancerous	B-Cancer
phenotypes	O
.	O

Matrix	B-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
activity	O
and	O
immunohistochemical	O
profile	O
of	O
matrix	B-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
-	B-Gene_or_gene_product
9	I-Gene_or_gene_product
and	O
tissue	B-Gene_or_gene_product
inhibitor	I-Gene_or_gene_product
of	I-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
during	O
human	B-Organism
dermal	B-Pathological_formation
wound	I-Pathological_formation
healing	O
.	O

Proteolytic	O
activity	O
is	O
required	O
for	O
the	O
turnover	O
of	O
the	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
during	O
wound	B-Pathological_formation
healing	O
.	O

Matrix	B-Gene_or_gene_product
metalloproteinases	I-Gene_or_gene_product
can	O
collectively	O
cleave	O
all	O
components	O
of	O
the	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
,	O
with	O
the	O
endogenous	O
tissue	B-Gene_or_gene_product
inhibitor	I-Gene_or_gene_product
of	I-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
regulating	O
their	O
activity	O
.	O

Breast	B-Tissue
tissue	I-Tissue
taken	O
at	O
varying	O
postoperative	O
times	O
(	O
n	O
=	O
92	O
)	O
or	O
during	O
surgery	O
(	O
controls	O
,	O
n	O
=	O
17	O
)	O
,	O
was	O
used	O
to	O
investigate	O
the	O
temporal	O
and	O
spatial	O
activity	O
of	O
matrix	B-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
-	B-Gene_or_gene_product
9	I-Gene_or_gene_product
and	O
tissue	B-Gene_or_gene_product
inhibitor	I-Gene_or_gene_product
of	I-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
during	O
human	B-Organism
wound	B-Pathological_formation
healing	O
.	O

Matrix	B-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
activity	O
,	O
determined	O
using	O
a	O
quenched	O
fluorescence	O
substrate	O
assay	O
,	O
increased	O
during	O
early	O
healing	O
(	O
3	O
-	O
8	O
weeks	O
)	O
compared	O
to	O
controls	O
,	O
and	O
then	O
decreased	O
between	O
24	O
and	O
36	O
weeks	O
after	O
surgery	O
(	O
p	O
less	O
than	O
0	O
.	O
05	O
until	O
24	O
weeks	O
,	O
Mann	O
-	O
Whitney	O
U	O
-	O
test	O
)	O
.	O

Immunohistochemistry	O
scores	O
for	O
matrix	B-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
expression	O
were	O
significantly	O
elevated	O
compared	O
to	O
controls	O
in	O
scar	B-Cell
endothelial	I-Cell
cells	I-Cell
and	O
fibroblasts	B-Cell
from	O
2	O
until	O
12	O
and	O
20	O
weeks	O
,	O
respectively	O
.	O

Matrix	B-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
staining	O
was	O
observed	O
exclusively	O
in	O
fibroblasts	B-Cell
,	O
reaching	O
maximum	O
levels	O
8	O
-	O
12	O
weeks	O
after	O
surgery	O
,	O
decreasing	O
by	O
1	O
.	O
5	O
years	O
but	O
remaining	O
significantly	O
increased	O
.	O

Tissue	B-Gene_or_gene_product
inhibitor	I-Gene_or_gene_product
of	I-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
staining	O
was	O
relatively	O
sparse	O
but	O
was	O
significantly	O
increased	O
until	O
8	O
weeks	O
after	O
surgery	O
.	O

These	O
results	O
show	O
that	O
matrix	B-Gene_or_gene_product
metalloproteinases	I-Gene_or_gene_product
are	O
present	O
at	O
elevated	O
levels	O
during	O
early	O
wound	B-Pathological_formation
healing	O
,	O
when	O
angiogenesis	O
occurs	O
,	O
and	O
suggest	O
that	O
matrix	B-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
may	O
play	O
a	O
significant	O
role	O
.	O

The	O
later	O
expression	O
of	O
matrix	B-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
-	B-Gene_or_gene_product
9	I-Gene_or_gene_product
in	O
fibroblasts	B-Cell
suggests	O
a	O
role	O
in	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
remodeling	O
.	O

Convergence	O
of	O
p53	B-Gene_or_gene_product
and	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
signaling	O
networks	O
.	O

p53	B-Gene_or_gene_product
is	O
a	O
protein	O
with	O
many	O
talents	O
.	O

One	O
of	O
the	O
most	O
fundamental	O
is	O
the	O
ability	O
to	O
act	O
as	O
essential	O
growth	O
checkpoint	O
that	O
protects	O
cells	B-Cell
against	O
cellular	B-Cell
transformation	O
.	O

p53	B-Gene_or_gene_product
does	O
so	O
through	O
the	O
induction	O
of	O
genes	O
leading	O
to	O
growth	O
arrest	O
or	O
apoptosis	O
.	O

Most	O
of	O
the	O
studies	O
focusing	O
on	O
the	O
mechanisms	O
of	O
p53	B-Gene_or_gene_product
activity	O
have	O
been	O
performed	O
in	O
cultured	O
cells	B-Cell
upon	O
treatment	O
with	O
well	O
-	O
established	O
p53	B-Gene_or_gene_product
-	O
activating	O
inputs	O
,	O
such	O
as	O
high	O
doses	O
of	O
radiations	O
,	O
DNA	B-Cellular_component
-	O
damaging	O
drugs	O
and	O
activated	O
oncogenes	O
.	O

However	O
,	O
how	O
the	O
tumor	B-Cancer
suppressive	O
functions	O
of	O
p53	B-Gene_or_gene_product
become	O
concerted	O
with	O
the	O
extracellular	B-Immaterial_anatomical_entity
cues	O
arriving	O
at	O
the	O
cell	B-Cellular_component
surface	I-Cellular_component
during	O
tissue	B-Tissue
homeostasis	O
,	O
remains	O
largely	O
unknown	O
.	O

Intriguingly	O
,	O
two	O
recent	O
papers	O
have	O
shed	O
new	O
light	O
into	O
this	O
unexplored	O
field	O
,	O
indicating	O
that	O
p53	B-Gene_or_gene_product
plays	O
a	O
key	O
role	O
in	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
-	O
induced	O
growth	O
arrest	O
and	O
,	O
unexpectedly	O
,	O
in	O
the	O
developmental	O
effects	O
of	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
in	O
early	O
embryos	B-Developing_anatomical_structure
.	O

Here	O
we	O
review	O
and	O
comment	O
on	O
these	O
findings	O
and	O
on	O
their	O
implications	O
for	O
cancer	B-Cancer
biology	O
.	O

Enhanced	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
expression	O
improves	O
smooth	B-Cell
muscle	I-Cell
cell	I-Cell
engraftment	O
after	O
cell	B-Cell
transplantation	O
.	O

The	O
functional	O
benefit	O
of	O
cell	B-Cell
transplantation	O
after	O
a	O
myocardial	B-Multi-tissue_structure
infarction	O
is	O
diminished	O
by	O
early	O
cell	B-Cell
losses	O
.	O

IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
enhances	O
cell	B-Cell
proliferation	O
and	O
survival	O
.	O

We	O
hypothesized	O
that	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
transfected	O
smooth	B-Cell
muscle	I-Cell
cells	I-Cell
(	O
SMCs	B-Cell
)	O
would	O
enhance	O
cell	B-Cell
survival	O
and	O
improve	O
engraftment	O
after	O
cell	B-Cell
transplantation	O
.	O

The	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
gene	O
was	O
transfected	O
into	O
male	O
SMCs	B-Cell
and	O
compared	O
with	O
SMCs	B-Cell
transfected	O
with	O
a	O
plasmid	O
vector	O
(	O
vector	O
control	O
)	O
and	O
nontransfected	O
SMCs	B-Cell
(	O
cell	B-Cell
control	O
)	O
.	O

IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
mRNA	O
(	O
n	O
=	O
10	O
/	O
group	O
)	O
and	O
protein	O
levels	O
(	O
n	O
=	O
6	O
/	O
group	O
)	O
were	O
higher	O
(	O
P	O
less	O
than	O
0	O
.	O
05	O
for	O
all	O
groups	O
)	O
at	O
3	O
,	O
7	O
,	O
and	O
14	O
days	O
compared	O
with	O
controls	O
.	O

VEGF	B-Gene_or_gene_product
was	O
also	O
increased	O
in	O
parallel	O
to	O
enhanced	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
expression	O
.	O

IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
transfected	O
cells	B-Cell
demonstrated	O
greater	O
cell	B-Cell
proliferation	O
,	O
stimulated	O
angiogenesis	O
,	O
and	O
decreased	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
activity	O
after	O
simulated	O
ischemia	O
and	O
reperfusion	O
(	O
P	O
less	O
than	O
0	O
.	O
05	O
for	O
all	O
groups	O
compared	O
with	O
vector	O
or	O
cell	B-Cell
controls	O
)	O
.	O

A	O
uniform	O
left	B-Multi-tissue_structure
ventricular	I-Multi-tissue_structure
injury	O
was	O
produced	O
in	O
female	O
rats	B-Organism
using	O
a	O
cryoprobe	O
.	O

Three	O
weeks	O
later	O
,	O
2	O
x	O
10	O
(	O
6	O
)	O
cells	B-Cell
from	O
three	O
groups	O
were	O
implanted	O
into	O
the	O
scar	B-Pathological_formation
.	O

One	O
week	O
later	O
,	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
transfected	O
SMCs	B-Cell
had	O
increased	O
myocardial	B-Multi-tissue_structure
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
VEGF	B-Gene_or_gene_product
levels	O
,	O
increased	O
Bcl2	B-Gene_or_gene_product
expression	O
,	O
limited	O
cell	B-Cell
apoptosis	O
,	O
and	O
enhanced	O
vessel	B-Multi-tissue_structure
formation	O
in	O
the	O
myocardial	B-Pathological_formation
scar	I-Pathological_formation
compared	O
with	O
the	O
two	O
control	O
groups	O
(	O
P	O
less	O
than	O
0	O
.	O
05	O
for	O
all	O
groups	O
)	O
.	O

The	O
proportion	O
of	O
SMCs	B-Cell
surviving	O
in	O
the	O
implanted	O
region	O
was	O
greater	O
(	O
P	O
less	O
than	O
0	O
.	O
05	O
)	O
in	O
the	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
transfected	O
group	O
than	O
in	O
the	O
vector	O
or	O
cell	B-Cell
controls	O
.	O

Gene	O
enhancement	O
with	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
improved	O
donor	B-Cell
cell	I-Cell
proliferation	O
,	O
survival	O
,	O
and	O
engraftment	O
after	O
cell	B-Cell
transplantation	O
,	O
perhaps	O
mediated	O
by	O
enhanced	O
angiogenesis	O
and	O
reduced	O
apoptosis	O
.	O

[	O
Altered	O
expression	O
of	O
PTEN	B-Gene_or_gene_product
gene	O
and	O
LOH	O
of	O
its	O
epigenetic	O
microsatellite	O
in	O
gastric	B-Cancer
carcinoma	I-Cancer
]	O
.	O

OBJECTIVE	O
:	O
To	O
investigate	O
the	O
expression	O
of	O
PTEN	B-Gene_or_gene_product
and	O
loss	O
of	O
heterozygosity	O
(	O
LOH	O
)	O
of	O
its	O
epigenetic	O
microsatellite	O
in	O
gastric	B-Cancer
carcinoma	I-Cancer
and	O
explore	O
their	O
roles	O
in	O
progression	O
of	O
gastric	B-Cancer
carcinoma	I-Cancer
.	O

METHODS	O
:	O
LOH	O
of	O
epigenetic	O
microsatellites	O
of	O
PTEN	B-Gene_or_gene_product
(	O
D10S541	O
,	O
D10S583	O
and	O
D10S1687	O
)	O
in	O
advanced	O
gastric	B-Cancer
cancer	I-Cancer
was	O
detected	O
by	O
PCR	O
-	O
SSCP	O
.	O

Expression	O
of	O
PTEN	B-Gene_or_gene_product
mRNA	O
and	O
protein	O
in	O
normal	O
gastric	B-Multi-tissue_structure
mucosa	I-Multi-tissue_structure
and	O
gastric	B-Cancer
cancer	I-Cancer
was	O
evaluated	O
by	O
RT	O
-	O
PCR	O
and	O
SABC	O
immunohistochemistry	O
,	O
respectively	O
.	O

The	O
relationship	O
between	O
expression	O
of	O
PTEN	B-Gene_or_gene_product
mRNA	O
and	O
protein	O
and	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
or	O
LOH	O
of	O
microsatellites	O
was	O
discussed	O
.	O

RESULTS	O
:	O
LOH	O
of	O
D10S541	O
,	O
D10S583	O
and	O
D10S1687	O
was	O
found	O
in	O
37	O
.	O
5	O
%	O
(	O
21	O
/	O
56	O
)	O
of	O
advanced	O
gastric	B-Cancer
cancers	I-Cancer
.	O

The	O
positive	O
rates	O
of	O
PTEN	B-Gene_or_gene_product
mRNA	O
expression	O
were	O
80	O
.	O
4	O
%	O
(	O
45	O
/	O
56	O
)	O
,	O
45	O
.	O
5	O
%	O
(	O
5	O
/	O
11	O
)	O
and	O
32	O
.	O
1	O
%	O
(	O
18	O
/	O
56	O
)	O
in	O
normal	O
mucosa	B-Multi-tissue_structure
,	O
early	O
and	O
advanced	O
gastric	B-Cancer
carcinomas	I-Cancer
,	O
respectively	O
,	O
while	O
78	O
.	O
6	O
%	O
(	O
44	O
/	O
56	O
)	O
,	O
44	O
.	O
5	O
%	O
(	O
5	O
/	O
11	O
)	O
and	O
28	O
.	O
6	O
%	O
(	O
16	O
/	O
56	O
)	O
at	O
the	O
protein	O
level	O
.	O

PTEN	B-Gene_or_gene_product
mRNA	O
and	O
protein	O
were	O
less	O
frequently	O
expressed	O
in	O
early	O
and	O
advanced	O
gastric	B-Cancer
carcinomas	I-Cancer
than	O
that	O
in	O
normal	O
gastric	B-Multi-tissue_structure
mucosa	I-Multi-tissue_structure
(	O
P	O
<	O
0	O
.	O
05	O
)	O
.	O

There	O
was	O
positive	O
correlation	O
between	O
PTEN	B-Gene_or_gene_product
mRNA	O
expression	O
and	O
LOH	O
of	O
microsatellites	O
in	O
advanced	O
gastric	B-Cancer
carcinomas	I-Cancer
.	O

PTEN	B-Gene_or_gene_product
protein	O
expression	O
paralleled	O
with	O
its	O
mRNA	O
expression	O
(	O
P	O
<	O
0	O
.	O
05	O
)	O
.	O

The	O
expression	O
of	O
PTEN	B-Gene_or_gene_product
mRNA	O
and	O
protein	O
was	O
negatively	O
correlated	O
with	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
of	O
advanced	O
gastric	B-Cancer
carcinomas	I-Cancer
(	O
P	O
<	O
0	O
.	O
05	O
)	O
.	O

CONCLUSION	O
:	O
Down	O
-	O
regulated	O
expression	O
of	O
PTEN	B-Gene_or_gene_product
gene	O
is	O
found	O
in	O
different	O
stages	O
of	O
gastric	B-Cancer
carcinoma	I-Cancer
,	O
and	O
is	O
closely	O
correlated	O
with	O
LOH	O
of	O
its	O
epigenetic	O
microsatellites	O
,	O
which	O
probably	O
is	O
its	O
underlying	O
molecular	O
mechanisms	O
.	O

It	O
suggests	O
that	O
altered	O
PTEN	B-Gene_or_gene_product
gene	O
contributes	O
to	O
tumorigenesis	O
and	O
progression	O
of	O
gastric	B-Cancer
carcinomas	I-Cancer
.	O

Granulocyte	B-Gene_or_gene_product
-	I-Gene_or_gene_product
macrophage	I-Gene_or_gene_product
colony	I-Gene_or_gene_product
-	I-Gene_or_gene_product
stimulating	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
and	O
interleukin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
enhance	O
the	O
incorporation	O
of	O
cytosine	B-Simple_chemical
arabinoside	I-Simple_chemical
into	O
the	O
DNA	B-Cellular_component
of	O
leukemic	B-Cell
blasts	I-Cell
and	O
the	O
cytotoxic	O
effect	O
on	O
clonogenic	B-Cell
cells	I-Cell
from	O
patients	B-Organism
with	O
acute	B-Cancer
myeloid	I-Cancer
leukemia	I-Cancer
.	O

In	O
the	O
present	O
study	O
the	O
effects	O
of	O
the	O
48	O
-	O
hour	O
administration	O
of	O
granulocyte	B-Gene_or_gene_product
-	I-Gene_or_gene_product
macrophage	I-Gene_or_gene_product
colony	I-Gene_or_gene_product
-	I-Gene_or_gene_product
stimulating	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
GM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
)	O
(	O
100	O
U	O
/	O
mL	O
)	O
or	O
interleukin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
(	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
)	O
(	O
100	O
U	O
/	O
mL	O
)	O
on	O
the	O
proliferative	O
activity	O
of	O
leukemic	B-Cell
cells	I-Cell
and	O
on	O
the	O
intracellular	B-Immaterial_anatomical_entity
metabolism	O
and	O
cytotoxic	O
efficacy	O
of	O
a	O
subsequent	O
12	O
-	O
hour	O
application	O
of	O
cytosine	B-Simple_chemical
arabinoside	I-Simple_chemical
(	O
ara	B-Simple_chemical
-	I-Simple_chemical
C	I-Simple_chemical
)	O
at	O
doses	O
of	O
0	O
.	O
1	O
,	O
1	O
.	O
0	O
,	O
10	O
.	O
0	O
,	O
and	O
100	O
.	O
0	O
mumol	O
/	O
L	O
were	O
evaluated	O
on	O
bone	B-Cell
marrow	I-Cell
cells	I-Cell
from	O
17	O
patients	B-Organism
with	O
acute	B-Cancer
myeloid	I-Cancer
leukemia	I-Cancer
.	O

After	O
GM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
or	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
,	O
a	O
1	O
.	O
2	O
-	O
to	O
2	O
.	O
4	O
-	O
fold	O
increase	O
in	O
S	O
-	O
phase	O
cells	B-Cell
was	O
observed	O
in	O
nine	O
of	O
14	O
GM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
and	O
seven	O
of	O
11	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
cases	O
.	O

3H	B-Simple_chemical
-	I-Simple_chemical
Cytosine	I-Simple_chemical
arabinoside	I-Simple_chemical
incorporation	O
into	O
the	O
DNA	B-Cellular_component
was	O
enhanced	O
1	O
.	O
33	O
-	O
to	O
18	O
.	O
3	O
-	O
fold	O
over	O
respective	O
controls	O
in	O
14	O
of	O
17	O
patients	B-Organism
.	O

While	O
in	O
control	O
specimens	B-Cell
are	O
ara	B-Simple_chemical
-	I-Simple_chemical
C	I-Simple_chemical
dose	O
-	O
dependent	O
increase	O
in	O
3H	B-Simple_chemical
-	I-Simple_chemical
ara	I-Simple_chemical
-	I-Simple_chemical
C	I-Simple_chemical
uptake	O
was	O
accompanied	O
by	O
a	O
corresponding	O
rise	O
in	O
intracellular	B-Immaterial_anatomical_entity
ara	B-Simple_chemical
-	I-Simple_chemical
C	I-Simple_chemical
-	I-Simple_chemical
5	I-Simple_chemical
'	I-Simple_chemical
triphosphate	I-Simple_chemical
(	O
ara	B-Simple_chemical
-	I-Simple_chemical
CTP	I-Simple_chemical
)	O
levels	O
,	O
ara	B-Simple_chemical
-	I-Simple_chemical
CTP	I-Simple_chemical
concentrations	O
were	O
not	O
increased	O
after	O
GM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
or	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
exposure	O
,	O
resulting	O
in	O
a	O
higher	O
ara	B-Simple_chemical
-	I-Simple_chemical
C	I-Simple_chemical
to	O
ara	B-Simple_chemical
-	I-Simple_chemical
CTP	I-Simple_chemical
ratio	O
over	O
controls	O
.	O

This	O
finding	O
may	O
be	O
explained	O
by	O
a	O
stimulatory	O
effect	O
of	O
GM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
and	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
on	O
ara	B-Simple_chemical
-	I-Simple_chemical
C	I-Simple_chemical
phosphorylating	O
enzymes	O
and	O
a	O
more	O
rapid	O
incorporation	O
of	O
ara	B-Simple_chemical
-	I-Simple_chemical
CTP	I-Simple_chemical
into	O
the	O
DNA	B-Cellular_component
of	O
leukemic	B-Cell
blasts	I-Cell
.	O

These	O
effects	O
translated	O
into	O
a	O
2	O
.	O
2	O
-	O
to	O
229	O
.	O
0	O
-	O
fold	O
increase	O
in	O
the	O
cytotoxic	O
activity	O
of	O
ara	B-Simple_chemical
-	I-Simple_chemical
C	I-Simple_chemical
against	O
clonogenic	B-Cell
leukemic	I-Cell
cells	I-Cell
after	O
GM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
or	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
pretreatment	O
.	O

Hence	O
,	O
GM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
and	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
enhance	O
the	O
intracellular	B-Immaterial_anatomical_entity
metabolism	O
of	O
ara	B-Simple_chemical
-	I-Simple_chemical
C	I-Simple_chemical
and	O
its	O
incorporation	O
into	O
the	O
DNA	B-Cellular_component
of	O
leukemic	B-Cell
cells	I-Cell
leading	O
to	O
a	O
higher	O
antileukemic	O
activity	O
of	O
ara	B-Simple_chemical
-	I-Simple_chemical
C	I-Simple_chemical
on	O
clonogenic	B-Cell
leukemic	I-Cell
cells	I-Cell
(	O
CFU	B-Cell
-	I-Cell
L	I-Cell
)	O
.	O

Silencing	O
of	O
monocarboxylate	B-Gene_or_gene_product
transporters	I-Gene_or_gene_product
via	O
small	O
interfering	O
ribonucleic	O
acid	O
inhibits	O
glycolysis	O
and	O
induces	O
cell	B-Cell
death	O
in	O
malignant	B-Cancer
glioma	I-Cancer
:	O
an	O
in	O
vitro	O
study	O
.	O

OBJECTIVE	O
:	O
Dependence	O
on	O
glycolysis	O
is	O
a	O
hallmark	O
of	O
malignant	B-Cancer
tumors	I-Cancer
.	O

As	O
a	O
consequence	O
,	O
these	O
tumors	B-Cancer
generate	O
more	O
lactate	B-Simple_chemical
,	O
which	O
is	O
effluxed	O
from	O
cells	B-Cell
by	O
monocarboxylate	B-Gene_or_gene_product
transporters	I-Gene_or_gene_product
(	O
MCTs	B-Gene_or_gene_product
)	O
.	O

We	O
hypothesized	O
that	O
1	O
)	O
MCT	B-Gene_or_gene_product
expression	O
in	O
malignant	B-Cancer
tumors	I-Cancer
may	O
differ	O
from	O
normal	B-Tissue
tissue	I-Tissue
in	O
quantity	O
,	O
isoform	O
,	O
or	O
both	O
;	O
and	O
2	O
)	O
silencing	O
MCT	B-Gene_or_gene_product
expression	O
would	O
induce	O
intracellular	B-Immaterial_anatomical_entity
acidification	O
,	O
resulting	O
in	O
decreased	O
proliferation	O
and	O
/	O
or	O
increased	O
cell	B-Cell
death	O
.	O

METHODS	O
:	O
We	O
quantified	O
expression	O
of	O
MCT	B-Gene_or_gene_product
isoforms	O
in	O
human	B-Organism
glioblastoma	B-Cell
multiforme	I-Cell
and	O
glioma	B-Cell
-	I-Cell
derived	I-Cell
cells	I-Cell
lines	I-Cell
by	O
Western	O
blot	O
analysis	O
.	O

MCTs	B-Gene_or_gene_product
that	O
were	O
abundant	O
or	O
specific	O
to	O
glioma	B-Cancer
then	O
were	O
targeted	O
in	O
the	O
model	O
U	B-Cell
-	I-Cell
87	I-Cell
MG	I-Cell
glioma	I-Cell
cell	I-Cell
line	I-Cell
via	O
small	O
interfering	O
ribonucleic	O
acid	O
-	O
mediated	O
gene	O
silencing	O
and	O
tested	O
for	O
inhibition	O
of	O
lactate	B-Simple_chemical
efflux	O
,	O
intracellular	B-Immaterial_anatomical_entity
pH	O
changes	O
,	O
reduced	O
proliferation	O
,	O
and	O
/	O
or	O
induction	O
of	O
cell	B-Cell
death	O
.	O

RESULTS	O
:	O
MCT	B-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
2	B-Gene_or_gene_product
were	O
the	O
primary	O
isoforms	O
expressed	O
in	O
human	B-Organism
glioblastoma	B-Cell
multiforme	I-Cell
and	O
glioma	B-Cell
-	I-Cell
derived	I-Cell
cell	I-Cell
lines	I-Cell
.	O

In	O
contrast	O
,	O
MCT	B-Gene_or_gene_product
3	I-Gene_or_gene_product
was	O
the	O
predominantly	O
expressed	O
isoform	O
in	O
normal	O
brain	B-Organ
.	O

Small	O
interfering	O
ribonucleic	O
acid	O
specific	O
for	O
MCT	B-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
2	B-Gene_or_gene_product
reduced	O
expression	O
of	O
these	O
isoforms	O
in	O
U	B-Cell
-	I-Cell
87	I-Cell
MG	I-Cell
cells	I-Cell
to	O
barely	O
detectable	O
levels	O
and	O
reduced	O
lactate	B-Simple_chemical
efflux	O
by	O
30	O
%	O
individually	O
and	O
85	O
%	O
in	O
combination	O
,	O
with	O
a	O
concomitant	O
decrease	O
of	O
intracellular	B-Immaterial_anatomical_entity
pH	O
by	O
0	O
.	O
6	O
units	O
(	O
a	O
fourfold	O
increase	O
in	O
intracellular	B-Immaterial_anatomical_entity
H	O
(	O
+	O
)	O
)	O
.	O

Prolonged	O
silencing	O
of	O
both	O
MCTs	B-Gene_or_gene_product
reduced	O
viability	O
by	O
75	O
%	O
individually	O
and	O
92	O
%	O
in	O
combination	O
,	O
as	O
measured	O
by	O
both	O
phenotypic	O
and	O
flow	O
cytometric	O
analyses	O
.	O

CONCLUSION	O
:	O
MCT	B-Gene_or_gene_product
targeting	O
significantly	O
reduced	O
the	O
viability	O
of	O
U	B-Cell
-	I-Cell
87	I-Cell
MG	I-Cell
cells	I-Cell
mediated	O
by	O
both	O
apoptosis	O
and	O
necrosis	O
.	O

This	O
indicates	O
that	O
the	O
strategy	O
may	O
be	O
a	O
useful	O
therapeutic	O
avenue	O
for	O
treatment	O
of	O
patients	B-Organism
with	O
malignant	B-Cancer
glioma	I-Cancer
.	O

Hypoxia	O
-	O
responsive	O
element	O
-	O
mediated	O
soluble	O
Tie2	B-Gene_or_gene_product
vector	O
exhibits	O
an	O
anti	O
-	O
angiogenic	O
activity	O
in	O
vitro	O
under	O
hypoxic	O
condition	O
.	O

Hypoxia	B-Gene_or_gene_product
-	I-Gene_or_gene_product
inducible	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
is	O
one	O
of	O
the	O
key	O
mammalian	O
transcription	O
factors	O
and	O
shows	O
increased	O
levels	O
in	O
both	O
protein	O
stability	O
and	O
intrinsic	O
transcriptional	O
activity	O
during	O
low	O
oxygen	B-Simple_chemical
tension	O
.	O

Hypoxia	O
-	O
activated	O
functional	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
protein	O
binds	O
to	O
hypoxia	O
-	O
responsive	O
elements	O
(	O
HRE	O
)	O
in	O
the	O
enhancers	O
of	O
several	O
genes	O
including	O
VEGF	B-Gene_or_gene_product
,	O
the	O
major	O
player	O
in	O
angiogenesis	O
,	O
and	O
initiates	O
their	O
mRNA	O
expression	O
.	O

The	O
molecular	O
mechanisms	O
regulating	O
the	O
gene	O
expression	O
under	O
hypoxic	O
conditions	O
could	O
increase	O
the	O
therapeutic	O
window	O
of	O
tumor	B-Cancer
-	O
specific	O
delivery	O
systems	O
.	O

In	O
this	O
study	O
,	O
to	O
examine	O
hypoxia	O
-	O
specific	O
production	O
of	O
anti	O
-	O
angiogenic	O
therapeutic	O
gene	O
,	O
we	O
constructed	O
5	O
copies	O
of	O
HRE	O
(	O
5xHRE	O
)	O
of	O
human	B-Organism
VEGF	B-Gene_or_gene_product
linked	O
to	O
soluble	O
Tie2	B-Gene_or_gene_product
(	O
sTie2	B-Gene_or_gene_product
)	O
driven	O
by	O
minimal	O
SV40	B-Gene_or_gene_product
promoter	O
(	O
5xHRE	O
/	O
SV40	B-Gene_or_gene_product
/	O
sTie2	B-Gene_or_gene_product
)	O
.	O

Our	O
data	O
showed	O
that	O
under	O
hypoxia	O
the	O
secreted	O
sTie2	B-Gene_or_gene_product
selectively	O
inhibited	O
tube	B-Tissue
formation	O
and	O
migration	O
capacities	O
of	O
endothelial	B-Cell
cells	I-Cell
in	O
vitro	O
.	O

Hence	O
,	O
we	O
propose	O
that	O
the	O
vector	O
system	O
,	O
5xHRE	O
/	O
SV40	B-Gene_or_gene_product
/	O
sTie2	B-Gene_or_gene_product
,	O
might	O
be	O
a	O
useful	O
tool	O
for	O
down	O
-	O
regulating	O
tumor	B-Cancer
angiogenesis	O
under	O
hypoxic	O
condition	O
.	O

Proteome	O
analysis	O
of	O
NIH3T3	B-Cell
cells	I-Cell
transformed	O
by	O
activated	O
Galpha12	B-Gene_or_gene_product
:	O
regulation	O
of	O
leukemia	B-Cancer
-	O
associated	O
protein	O
SET	B-Gene_or_gene_product
.	O

Galpha	B-Gene_or_gene_product
(	I-Gene_or_gene_product
12	I-Gene_or_gene_product
)	I-Gene_or_gene_product
,	O
the	O
alpha	O
-	O
subunit	O
of	O
the	O
G12	B-Gene_or_gene_product
family	O
of	O
heterotrimeric	B-Gene_or_gene_product
G	I-Gene_or_gene_product
proteins	I-Gene_or_gene_product
is	O
involved	O
in	O
the	O
regulation	O
of	O
cell	B-Cell
proliferation	O
and	O
neoplastic	O
transformation	O
.	O

GTPase	B-Gene_or_gene_product
-	O
deficient	O
,	O
constitutively	O
activated	O
mutant	O
of	O
Galpha	B-Gene_or_gene_product
(	I-Gene_or_gene_product
12	I-Gene_or_gene_product
)	I-Gene_or_gene_product
(	O
Galpha	B-Gene_or_gene_product
(	I-Gene_or_gene_product
12	I-Gene_or_gene_product
)	I-Gene_or_gene_product
Q229L	I-Gene_or_gene_product
or	O
Galpha	B-Gene_or_gene_product
(	I-Gene_or_gene_product
12	I-Gene_or_gene_product
)	I-Gene_or_gene_product
QL	I-Gene_or_gene_product
)	O
has	O
been	O
previously	O
shown	O
to	O
induce	O
oncogenic	O
transformation	O
of	O
NIH3T3	B-Cell
cells	I-Cell
promoting	O
serum	B-Organism_substance
-	O
and	O
anchorage	O
-	O
independent	O
growth	O
.	O

Reduced	O
growth	O
-	O
factor	O
dependent	O
,	O
autonomous	O
cell	B-Cell
growth	O
forms	O
a	O
critical	O
defining	O
point	O
at	O
which	O
a	O
normal	O
cell	B-Cell
turns	O
into	O
an	O
oncogenic	O
one	O
.	O

To	O
identify	O
the	O
underlying	O
mechanism	O
involved	O
in	O
such	O
growth	O
-	O
factor	O
/	O
serum	B-Organism_substance
independent	O
growth	O
of	O
Galpha	B-Gene_or_gene_product
(	I-Gene_or_gene_product
12	I-Gene_or_gene_product
)	I-Gene_or_gene_product
QL	I-Gene_or_gene_product
-	O
transformed	O
NIH3T3	B-Cell
,	O
we	O
carried	O
out	O
a	O
two	O
-	O
dimensional	O
differential	O
proteome	O
analysis	O
of	O
Galpha	B-Gene_or_gene_product
(	I-Gene_or_gene_product
12	I-Gene_or_gene_product
)	I-Gene_or_gene_product
QL	I-Gene_or_gene_product
-	O
transformed	O
NIH3T3	B-Cell
cells	I-Cell
and	O
cells	B-Cell
expressing	O
vector	O
control	O
.	O

This	O
analysis	O
revealed	O
a	O
total	O
of	O
22	O
protein	O
-	O
spots	O
whose	O
expression	O
was	O
altered	O
by	O
more	O
than	O
3	O
-	O
folds	O
.	O

Two	O
of	O
these	O
spots	O
were	O
identified	O
by	O
MALDI	O
-	O
MS	O
analysis	O
as	O
proliferating	B-Gene_or_gene_product
cell	I-Gene_or_gene_product
nuclear	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
(	O
PCNA	B-Gene_or_gene_product
)	O
and	O
myeloid	B-Cancer
-	I-Cancer
leukemia	I-Cancer
-	O
associated	O
SET	B-Gene_or_gene_product
protein	O
.	O

The	O
increased	O
expressions	O
of	O
these	O
proteins	O
in	O
Galpha	B-Gene_or_gene_product
(	I-Gene_or_gene_product
12	I-Gene_or_gene_product
)	I-Gene_or_gene_product
QL	I-Gene_or_gene_product
cells	O
were	O
validated	O
by	O
immunoblot	O
analysis	O
.	O

Furthermore	O
,	O
transient	O
transfection	O
studies	O
with	O
NIH3T3	B-Cell
cells	I-Cell
indicated	O
that	O
the	O
expression	O
of	O
activated	O
Galpha	B-Gene_or_gene_product
(	I-Gene_or_gene_product
12	I-Gene_or_gene_product
)	I-Gene_or_gene_product
readily	O
increased	O
the	O
expression	O
of	O
SET	B-Gene_or_gene_product
protein	O
by	O
24	O
h	O
.	O

As	O
SET	B-Gene_or_gene_product
has	O
been	O
previously	O
reported	O
to	O
be	O
an	O
inhibitor	O
of	O
phosphatase	O
PP2A	B-Gene_or_gene_product
,	O
the	O
nuclear	B-Cellular_component
phosphatase	O
activity	O
was	O
monitored	O
in	O
cells	B-Cell
expressing	O
activated	O
Galpha	B-Gene_or_gene_product
(	I-Gene_or_gene_product
12	I-Gene_or_gene_product
)	I-Gene_or_gene_product
.	O

Our	O
results	O
indicate	O
that	O
the	O
nuclear	B-Cellular_component
phosphatase	O
activity	O
is	O
inhibited	O
by	O
greater	O
than	O
50	O
%	O
in	O
Galpha	B-Gene_or_gene_product
(	I-Gene_or_gene_product
12	I-Gene_or_gene_product
)	I-Gene_or_gene_product
QL	I-Gene_or_gene_product
cells	O
compared	O
to	O
vector	O
control	O
cells	B-Cell
.	O

Thus	O
,	O
our	O
results	O
from	O
differential	O
proteome	O
analysis	O
presented	O
here	O
report	O
for	O
the	O
first	O
time	O
a	O
role	O
for	O
SET	B-Gene_or_gene_product
in	O
Galpha	B-Gene_or_gene_product
(	I-Gene_or_gene_product
12	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	O
mediated	O
signaling	O
pathways	O
and	O
a	O
role	O
for	O
Galpha	B-Gene_or_gene_product
(	I-Gene_or_gene_product
12	I-Gene_or_gene_product
)	I-Gene_or_gene_product
in	O
the	O
regulation	O
of	O
the	O
leukemia	B-Cancer
-	O
associated	O
SET	B-Gene_or_gene_product
-	O
protein	O
expression	O
.	O

Matrix	O
metalloproteinase	O
/	O
tissue	O
inhibitors	O
of	O
matrix	O
metalloproteinase	O
phenotype	O
identifies	O
poor	O
prognosis	O
colorectal	B-Cancer
cancers	I-Cancer
.	O

PURPOSE	O
:	O
The	O
matrix	O
metalloproteinases	O
(	O
MMPs	O
)	O
are	O
a	O
family	O
of	O
proteolytic	O
enzymes	O
involved	O
in	O
tumor	B-Cancer
invasion	O
;	O
several	O
individual	O
members	O
of	O
which	O
have	O
been	O
implicated	O
in	O
tumor	B-Cancer
prognosis	O
.	O

These	O
enzymes	O
and	O
their	O
physiologic	O
inhibitors	O
,	O
the	O
tissue	O
inhibitors	O
of	O
matrix	O
metalloproteinases	O
(	O
TIMPs	O
)	O
,	O
act	O
in	O
a	O
coordinated	O
manner	O
to	O
form	O
an	O
integrated	O
system	O
.	O

Therefore	O
,	O
to	O
understand	O
their	O
role	O
in	O
tumor	B-Cancer
invasion	O
,	O
it	O
is	O
necessary	O
to	O
evaluate	O
them	O
collectively	O
.	O

EXPERIMENTAL	O
DESIGN	O
:	O
In	O
this	O
study	O
all	O
of	O
the	O
major	O
members	O
of	O
the	O
matrix	O
metalloproteinase	O
(	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
,	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
7	I-Gene_or_gene_product
,	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
,	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
13	I-Gene_or_gene_product
,	O
MT1	B-Gene_or_gene_product
-	I-Gene_or_gene_product
MMP	I-Gene_or_gene_product
and	O
MT2	B-Gene_or_gene_product
-	I-Gene_or_gene_product
MMP	I-Gene_or_gene_product
)	O
/	O
tissue	O
inhibitor	O
of	O
matrix	O
metalloproteinase	O
(	O
TIMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
TIMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
and	O
TIMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
)	O
system	O
have	O
been	O
investigated	O
by	O
immunohistochemistry	O
in	O
a	O
series	O
(	O
n	O
=	O
90	O
)	O
of	O
stage	B-Cancer
III	I-Cancer
(	I-Cancer
Dukes	I-Cancer
'	I-Cancer
C	I-Cancer
)	I-Cancer
colorectal	I-Cancer
cancers	I-Cancer
.	O

An	O
immunohistochemical	O
score	O
based	O
on	O
the	O
intensity	O
of	O
immunoreactivity	O
and	O
proportion	O
of	O
immunoreactive	O
cells	B-Cell
was	O
established	O
for	O
each	O
MMP	O
and	O
TIMP	O
.	O

RESULTS	O
:	O
The	O
MMP	O
/	O
TIMP	O
profile	O
defined	O
by	O
hierarchical	O
cluster	O
analysis	O
of	O
the	O
immunohistochemical	O
score	O
identifies	O
a	O
distinct	O
group	O
of	O
colorectal	B-Cancer
cancers	I-Cancer
with	O
poor	O
prognosis	O
(	O
log	O
-	O
rank	O
test	O
,	O
12	O
.	O
22	O
,	O
P	O
=	O
0	O
.	O
0005	O
)	O
.	O

The	O
median	O
survival	O
time	O
of	O
patients	B-Organism
in	O
this	O
survival	O
group	O
was	O
18	O
months	O
compared	O
with	O
a	O
median	O
survival	O
of	O
49	O
months	O
in	O
the	O
"	O
good	O
"	O
survival	O
group	O
.	O

Multivariate	O
analysis	O
showed	O
that	O
this	O
profile	O
was	O
independently	O
the	O
most	O
significant	O
prognostic	O
factor	O
(	O
P	O
=	O
0	O
.	O
001	O
)	O
.	O

CONCLUSIONS	O
:	O
This	O
study	O
has	O
identified	O
that	O
the	O
MMP	O
/	O
TIMP	O
profile	O
is	O
an	O
independent	O
indicator	O
of	O
poor	O
prognosis	O
in	O
colorectal	B-Cancer
cancer	I-Cancer
.	O

2	B-Simple_chemical
-	I-Simple_chemical
methoxyestradiol	I-Simple_chemical
inhibits	O
hypoxia	B-Gene_or_gene_product
-	I-Gene_or_gene_product
inducible	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
,	O
tumor	B-Cancer
growth	O
,	O
and	O
angiogenesis	O
and	O
augments	O
paclitaxel	B-Simple_chemical
efficacy	O
in	O
head	B-Cancer
and	I-Cancer
neck	I-Cancer
squamous	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
.	O

PURPOSE	O
:	O
Head	B-Cancer
and	I-Cancer
neck	I-Cancer
squamous	I-Cancer
cell	I-Cancer
carcinomas	I-Cancer
have	O
been	O
reported	O
to	O
overexpress	O
hypoxia	B-Gene_or_gene_product
-	I-Gene_or_gene_product
inducible	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	I-Gene_or_gene_product
HIF	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
,	O
a	O
transcription	O
factor	O
that	O
promotes	O
expression	O
of	O
angiogenesis	O
factors	O
and	O
resistance	O
to	O
programmed	O
and	O
therapy	O
-	O
induced	O
cell	B-Cell
death	O
.	O

2	B-Simple_chemical
-	I-Simple_chemical
Methoxyestradiol	I-Simple_chemical
(	O
2ME2	B-Simple_chemical
)	O
is	O
a	O
natural	O
compound	O
with	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
inhibitory	O
activity	O
that	O
is	O
currently	O
being	O
evaluated	O
in	O
phase	O
1	O
and	O
2	O
clinical	O
trials	O
for	O
advanced	O
solid	B-Cancer
tumors	I-Cancer
and	O
multiple	O
myeloma	B-Cancer
.	O

To	O
our	O
knowledge	O
,	O
this	O
is	O
the	O
first	O
study	O
to	O
evaluate	O
the	O
effects	O
of	O
2ME2	B-Simple_chemical
in	O
head	B-Cancer
and	I-Cancer
neck	I-Cancer
squamous	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
.	O

EXPERIMENTAL	O
DESIGN	O
:	O
In	O
the	O
present	O
study	O
,	O
we	O
investigated	O
the	O
effects	O
of	O
2ME2	B-Simple_chemical
alone	O
and	O
in	O
combination	O
with	O
paclitaxel	B-Simple_chemical
,	O
an	O
active	O
agent	O
in	O
recurrent	O
or	O
advanced	O
head	B-Cancer
and	I-Cancer
neck	I-Cancer
squamous	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
.	O

RESULTS	O
:	O
2ME2	B-Simple_chemical
exhibited	O
antiproliferative	O
and	O
cytotoxic	O
effects	O
in	O
a	O
panel	O
of	O
five	O
head	B-Cell
and	I-Cell
neck	I-Cell
squamous	I-Cell
cell	I-Cell
carcinoma	I-Cell
cell	I-Cell
lines	I-Cell
in	O
the	O
0	O
.	O
5	O
to	O
10	O
micromol	O
/	O
L	O
range	O
,	O
including	O
induction	O
of	O
G2	O
-	O
M	O
blockade	O
,	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
/	O
7	B-Gene_or_gene_product
activation	O
,	O
and	O
apoptosis	O
at	O
48	O
hours	O
.	O

2ME2	B-Simple_chemical
resulted	O
in	O
decreased	O
nuclear	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
-	O
binding	O
activity	O
and	O
affected	O
the	O
expression	O
of	O
downstream	O
genes	O
,	O
such	O
as	O
bid	B-Gene_or_gene_product
,	O
a	O
proapoptotic	O
bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
family	O
member	O
,	O
and	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
,	O
a	O
proangiogenic	O
cytokine	O
.	O

The	O
up	O
-	O
regulation	O
of	O
Bid	B-Gene_or_gene_product
(	O
57	O
.	O
5	O
%	O
at	O
12	O
hours	O
,	O
P	O
less	O
than	O
0	O
.	O
0006	O
)	O
and	O
inhibition	O
of	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
secretion	O
(	O
57	O
.	O
7	O
%	O
at	O
24	O
hours	O
,	O
P	O
less	O
than	O
0	O
.	O
015	O
;	O
and	O
50	O
.	O
3	O
%	O
at	O
48	O
hours	O
,	O
P	O
less	O
than	O
0	O
.	O
0006	O
)	O
could	O
be	O
partially	O
attributed	O
to	O
the	O
effects	O
on	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
,	O
because	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
small	O
interfering	O
RNAs	O
produced	O
similar	O
effects	O
.	O

Finally	O
,	O
in	O
vivo	O
,	O
in	O
a	O
xenograft	B-Cancer
model	O
of	O
head	B-Cancer
and	I-Cancer
neck	I-Cancer
squamous	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
using	O
UM	B-Cell
-	I-Cell
SCC	I-Cell
-	I-Cell
11A	I-Cell
cells	I-Cell
,	O
2ME2	B-Simple_chemical
exhibited	O
antitumor	B-Cancer
and	O
antiangiogenic	O
activity	O
,	O
as	O
measured	O
by	O
CD31	B-Gene_or_gene_product
immunostaining	O
.	O

CONCLUSIONS	O
:	O
These	O
results	O
provide	O
support	O
for	O
the	O
use	O
of	O
2ME2	B-Simple_chemical
in	O
combination	O
with	O
paclitaxel	B-Simple_chemical
for	O
the	O
treatment	O
of	O
recurrent	O
or	O
advanced	O
head	B-Cancer
and	I-Cancer
neck	I-Cancer
squamous	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
.	O

HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2alpha	I-Gene_or_gene_product
expression	O
in	O
human	B-Organism
fetal	B-Cell
paraganglia	I-Cell
and	O
neuroblastoma	B-Cancer
:	O
relation	O
to	O
sympathetic	O
differentiation	O
,	O
glucose	B-Simple_chemical
deficiency	O
,	O
and	O
hypoxia	O
.	O

Solid	B-Cancer
tumors	I-Cancer
are	O
frequently	O
necrotic	O
and	O
hypoxic	O
due	O
to	O
poor	O
vascularization	O
.	O

Tumor	B-Cell
cells	I-Cell
adapt	O
to	O
hypoxia	O
by	O
modulating	O
their	O
phenotype	O
.	O

Key	O
players	O
in	O
this	O
process	O
are	O
the	O
hypoxia	B-Gene_or_gene_product
-	I-Gene_or_gene_product
inducible	I-Gene_or_gene_product
factors	I-Gene_or_gene_product
(	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
to	O
3alpha	B-Gene_or_gene_product
)	O
.	O

HIFs	B-Gene_or_gene_product
are	O
also	O
expressed	O
during	O
normal	O
development	O
;	O
for	O
example	O
,	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2alpha	I-Gene_or_gene_product
is	O
specifically	O
expressed	O
and	O
appears	O
to	O
be	O
involved	O
in	O
the	O
development	O
of	O
the	O
murine	B-Organism
sympathetic	B-Anatomical_system
nervous	I-Anatomical_system
system	I-Anatomical_system
(	O
SNS	B-Anatomical_system
)	O
.	O

Here	O
,	O
we	O
demonstrate	O
that	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2alpha	I-Gene_or_gene_product
protein	O
is	O
selectively	O
present	O
in	O
human	B-Organism
fetal	B-Developing_anatomical_structure
week	O
8	O
.	O
5	O
SNS	B-Cell
paraganglia	I-Cell
.	O

Neuroblastoma	B-Cancer
is	O
derived	O
from	O
SNS	B-Developing_anatomical_structure
precursors	I-Developing_anatomical_structure
.	O

In	O
a	O
subset	O
of	O
neuroblastomas	B-Cancer
,	O
a	O
spontaneous	O
neuronal	B-Cell
to	O
neuroendocrine	B-Cell
differentiation	O
occurs	O
in	O
areas	O
adjacent	O
to	O
necrotic	B-Tissue
zones	I-Tissue
.	O

As	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2alpha	I-Gene_or_gene_product
activity	O
has	O
been	O
associated	O
not	O
only	O
with	O
hypoxic	O
but	O
also	O
with	O
hypoglycemic	O
conditions	O
,	O
we	O
have	O
investigated	O
putative	O
effects	O
of	O
hypoxia	O
,	O
glucose	B-Simple_chemical
depletion	O
,	O
and	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2alpha	I-Gene_or_gene_product
on	O
the	O
neuroblastoma	B-Cancer
phenotype	O
.	O

HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2alpha	I-Gene_or_gene_product
was	O
detected	O
in	O
hypoxic	O
and	O
in	O
well	O
-	O
oxygenized	O
neuroblastoma	B-Cell
cells	I-Cell
and	O
tissue	B-Tissue
,	O
presumably	O
reflecting	O
their	O
embryonic	B-Developing_anatomical_structure
features	O
.	O

With	O
regard	O
to	O
differentiation	O
,	O
hypoxic	B-Cell
cells	I-Cell
lost	O
their	O
neuronal	B-Cell
/	O
neuroendocrine	B-Cell
features	O
and	O
gained	O
marker	O
gene	O
expression	O
associated	O
with	O
an	O
immature	O
,	O
neural	O
crest	O
-	O
like	O
phenotype	O
.	O

Low	O
glucose	B-Simple_chemical
potentiated	O
the	O
effect	O
of	O
hypoxia	O
.	O

These	O
findings	O
suggest	O
that	O
poorly	O
vascularized	O
neuroblastomas	B-Cancer
become	O
immature	O
and	O
maintain	O
a	O
more	O
aggressive	O
phenotype	O
,	O
which	O
possibly	O
could	O
involve	O
a	O
sustained	O
stabilization	O
and	O
activation	O
of	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2alpha	I-Gene_or_gene_product
.	O

Expression	O
and	O
purification	O
of	O
the	O
catalytic	O
domain	O
of	O
human	B-Organism
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
2	I-Gene_or_gene_product
for	O
inhibitor	O
screening	O
.	O

Vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
,	O
an	O
endothelial	B-Cell
cell	I-Cell
-	O
specific	O
mitogen	O
,	O
can	O
act	O
in	O
tumor	B-Cancer
-	O
induced	O
angiogenesis	O
by	O
binding	O
to	O
specific	O
receptors	O
on	O
the	O
surface	B-Cellular_component
of	O
endothelial	B-Cell
cells	I-Cell
.	O

One	O
such	O
receptor	O
,	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
/	O
KDR	B-Gene_or_gene_product
,	O
plays	O
a	O
key	O
role	O
in	O
VEGF	B-Gene_or_gene_product
-	O
induced	O
angiogenesis	O
.	O

Here	O
,	O
we	O
expressed	O
the	O
catalytic	O
domain	O
of	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
as	O
a	O
soluble	O
active	O
kinase	O
using	O
Bac	O
-	O
to	O
-	O
Bac	O
expression	O
system	O
,	O
and	O
investigated	O
correlations	O
between	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
activity	O
and	O
enzyme	O
concentration	O
,	O
ATP	B-Simple_chemical
concentration	O
,	O
substrate	O
concentration	O
and	O
divalent	O
cation	O
type	O
.	O

We	O
used	O
these	O
data	O
to	O
establish	O
a	O
convenient	O
,	O
effective	O
and	O
non	O
-	O
radioactive	O
ELISA	O
screening	O
technique	O
for	O
the	O
identification	O
and	O
evaluation	O
of	O
potential	O
inhibitors	O
for	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
kinase	O
.	O

We	O
screened	O
200	O
RTK	O
target	O
-	O
based	O
compounds	O
and	O
identified	O
one	O
(	O
TKI	B-Simple_chemical
-	I-Simple_chemical
31	I-Simple_chemical
)	O
that	O
potently	O
inhibited	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
kinase	O
activity	O
(	O
IC50	O
=	O
0	O
.	O
596	O
microM	O
)	O
.	O

Treatment	O
of	O
NIH3T3	B-Cell
/	I-Cell
KDR	I-Cell
cells	I-Cell
with	O
TKI	B-Simple_chemical
-	I-Simple_chemical
31	I-Simple_chemical
blocked	O
VEGF	B-Gene_or_gene_product
-	O
induced	O
phosphorylation	O
of	O
KDR	B-Gene_or_gene_product
in	O
a	O
dose	O
-	O
dependent	O
manner	O
.	O

Moreover	O
,	O
TKI	B-Simple_chemical
-	I-Simple_chemical
31	I-Simple_chemical
dose	O
-	O
dependently	O
suppressed	O
HUVEC	B-Tissue
tube	I-Tissue
formation	O
.	O

Thus	O
,	O
we	O
herein	O
report	O
a	O
novel	O
,	O
efficient	O
method	O
for	O
identifying	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
kinase	O
inhibitors	O
and	O
introduce	O
one	O
,	O
TKI	B-Simple_chemical
-	I-Simple_chemical
31	I-Simple_chemical
,	O
that	O
may	O
prove	O
to	O
be	O
a	O
useful	O
new	O
angiogenesis	O
inhibitor	O
.	O

Role	O
of	O
thrombogenic	B-Gene_or_gene_product
factors	I-Gene_or_gene_product
in	O
the	O
development	O
of	O
atherosclerosis	O
.	O

Hemostatic	B-Gene_or_gene_product
factors	I-Gene_or_gene_product
play	O
a	O
crucial	O
role	O
in	O
generating	O
thrombotic	B-Pathological_formation
plugs	I-Pathological_formation
at	O
sites	O
of	O
vascular	B-Multi-tissue_structure
damage	O
(	O
atherothrombosis	O
)	O
.	O

However	O
,	O
whether	O
hemostatic	B-Gene_or_gene_product
factors	I-Gene_or_gene_product
contribute	O
directly	O
or	O
indirectly	O
to	O
the	O
pathogenesis	O
of	O
atherosclerosis	O
remains	O
uncertain	O
.	O

Autopsy	O
studies	O
have	O
revealed	O
that	O
intimal	B-Tissue
thickening	O
represents	O
the	O
first	O
stage	O
of	O
atherosclerosis	O
and	O
that	O
lipid	B-Pathological_formation
-	I-Pathological_formation
rich	I-Pathological_formation
plaque	I-Pathological_formation
arises	O
from	O
such	O
lesions	B-Pathological_formation
.	O

Several	O
factors	O
contribute	O
to	O
the	O
start	O
of	O
intimal	B-Tissue
thickening	O
.	O

Platelets	B-Cell
release	O
several	O
growth	O
factors	O
and	O
bioactive	O
agents	O
that	O
play	O
a	O
central	O
role	O
in	O
development	O
of	O
not	O
only	O
thrombus	B-Pathological_formation
but	O
also	O
of	O
intimal	B-Tissue
thickening	O
.	O

We	O
have	O
been	O
investigating	O
which	O
coagulation	O
factors	O
simultaneously	O
,	O
or	O
subsequently	O
with	O
platelet	B-Cell
aggregation	O
,	O
participate	O
in	O
thrombus	B-Pathological_formation
formation	O
.	O

Tissue	B-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
TF	B-Gene_or_gene_product
)	O
is	O
an	O
essential	O
initiator	O
of	O
blood	B-Organism_substance
coagulation	O
that	O
is	O
expressed	O
in	O
various	O
stages	O
of	O
atherosclerotic	B-Pathological_formation
lesions	I-Pathological_formation
in	O
humans	B-Organism
and	O
other	O
animals	O
.	O

Factors	O
including	O
thrombin	B-Gene_or_gene_product
and	O
fibrin	B-Gene_or_gene_product
,	O
which	O
are	O
downstream	O
of	O
the	O
coagulation	O
cascade	O
activated	O
by	O
TF	B-Gene_or_gene_product
,	O
also	O
contribute	O
to	O
atherosclerosis	O
.	O

TF	B-Gene_or_gene_product
is	O
involved	O
in	O
cell	B-Cell
migration	O
,	O
embryogenesis	O
and	O
angiogenesis	O
.	O

Thus	O
TF	B-Gene_or_gene_product
,	O
in	O
addition	O
to	O
factors	O
downstream	O
of	O
the	O
coagulation	O
cascade	O
and	O
the	O
protease	B-Gene_or_gene_product
-	I-Gene_or_gene_product
activated	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
2	I-Gene_or_gene_product
activation	O
system	O
,	O
would	O
be	O
a	O
multifactorial	O
regulator	O
of	O
atherogenesis	O
.	O

Angiogenin	B-Gene_or_gene_product
is	O
up	O
-	O
regulated	O
in	O
the	O
nucleus	B-Cellular_component
and	O
cytoplasm	B-Organism_substance
in	O
human	B-Organism
primary	O
breast	B-Cancer
carcinoma	I-Cancer
and	O
is	O
associated	O
with	O
markers	O
of	O
hypoxia	O
but	O
not	O
survival	O
.	O

Angiogenin	B-Gene_or_gene_product
,	O
a	O
14	O
.	O
2	O
kD	O
polypeptide	O
that	O
was	O
originally	O
noted	O
for	O
its	O
angiogenic	O
activity	O
,	O
is	O
now	O
increasingly	O
recognized	O
to	O
have	O
a	O
multiplicity	O
of	O
biological	O
roles	O
in	O
both	O
physiological	O
and	O
pathological	O
conditions	O
.	O

In	O
breast	B-Cancer
cancer	I-Cancer
,	O
there	O
are	O
conflicting	O
studies	O
questioning	O
the	O
role	O
of	O
angiogenin	B-Gene_or_gene_product
.	O

Here	O
,	O
the	O
pattern	O
of	O
expression	O
of	O
angiogenin	B-Gene_or_gene_product
during	O
the	O
transition	O
from	O
normal	O
breast	B-Tissue
tissue	I-Tissue
to	O
ductal	B-Cancer
carcinoma	I-Cancer
in	O
situ	O
and	O
invasive	O
carcinoma	B-Cancer
is	O
reported	O
together	O
with	O
the	O
correlates	O
between	O
the	O
level	O
of	O
angiogenin	B-Gene_or_gene_product
in	O
239	O
invasive	O
carcinomas	B-Cancer
and	O
standard	O
clinicopathological	O
parameters	O
,	O
hypoxia	B-Gene_or_gene_product
-	I-Gene_or_gene_product
inducible	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	I-Gene_or_gene_product
HIF	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
and	O
the	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
target	O
gene	O
DEC	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

This	O
study	O
shows	O
that	O
angiogenin	B-Gene_or_gene_product
expression	O
is	O
up	O
-	O
regulated	O
in	O
the	O
cytoplasmic	B-Organism_substance
and	O
nuclear	B-Cellular_component
compartments	O
in	O
in	O
situ	O
carcinoma	B-Cancer
and	O
invasive	O
carcinoma	B-Cancer
compared	O
with	O
normal	O
breast	B-Tissue
tissue	I-Tissue
and	O
that	O
angiogenin	B-Gene_or_gene_product
expression	O
in	O
invasive	O
carcinomas	B-Cancer
is	O
significantly	O
positively	O
associated	O
with	O
high	O
tumour	B-Cancer
grade	O
(	O
p	O
=	O
0	O
.	O
03	O
)	O
,	O
positive	O
oestrogen	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
(	O
ER	B-Gene_or_gene_product
)	O
status	O
(	O
p	O
=	O
0	O
.	O
01	O
)	O
,	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
(	O
p	O
=	O
0	O
.	O
001	O
)	O
and	O
DEC	B-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
p	O
=	O
0	O
.	O
001	O
)	O
,	O
but	O
not	O
with	O
patient	B-Organism
age	O
(	O
p	O
=	O
0	O
.	O
8	O
)	O
,	O
tumour	B-Cancer
size	O
(	O
p	O
=	O
0	O
.	O
25	O
)	O
,	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
status	O
(	O
p	O
=	O
0	O
.	O
69	O
)	O
,	O
epidermal	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
(	O
p	O
=	O
0	O
.	O
56	O
)	O
or	O
microvessel	B-Tissue
density	O
(	O
p	O
=	O
0	O
.	O
32	O
)	O
.	O

No	O
difference	O
in	O
relapse	O
-	O
free	O
(	O
p	O
=	O
0	O
.	O
26	O
)	O
or	O
overall	O
(	O
p	O
=	O
0	O
.	O
63	O
)	O
survival	O
was	O
observed	O
in	O
patients	B-Organism
stratified	O
by	O
angiogenin	B-Gene_or_gene_product
expression	O
.	O

This	O
study	O
suggests	O
that	O
angiogenin	B-Gene_or_gene_product
may	O
be	O
important	O
in	O
breast	B-Cancer
cancer	I-Cancer
progression	O
and	O
that	O
,	O
through	O
its	O
relationship	O
with	O
ER	B-Gene_or_gene_product
,	O
it	O
may	O
be	O
a	O
target	O
for	O
tamoxifen	B-Simple_chemical
.	O

Hereditary	O
paraganglioma	B-Cancer
/	O
pheochromocytoma	B-Cancer
and	O
inherited	O
succinate	B-Gene_or_gene_product
dehydrogenase	I-Gene_or_gene_product
deficiency	O
.	O

Mitochondrial	B-Cellular_component
complex	B-Gene_or_gene_product
II	I-Gene_or_gene_product
,	O
or	O
succinate	B-Gene_or_gene_product
dehydrogenase	I-Gene_or_gene_product
,	O
is	O
a	O
key	O
enzymatic	O
complex	O
involved	O
in	O
both	O
the	O
tricarboxylic	B-Simple_chemical
acid	I-Simple_chemical
(	O
TCA	B-Simple_chemical
)	O
cycle	O
and	O
oxidative	O
phosphorylation	O
as	O
part	O
of	O
the	O
mitochondrial	B-Cellular_component
respiratory	O
chain	O
.	O

Germline	O
succinate	B-Gene_or_gene_product
dehydrogenase	I-Gene_or_gene_product
subunit	I-Gene_or_gene_product
A	I-Gene_or_gene_product
(	O
SDHA	B-Gene_or_gene_product
)	O
mutations	O
have	O
been	O
reported	O
in	O
a	O
few	O
patients	B-Organism
with	O
a	O
classical	O
mitochondrial	B-Cellular_component
neurodegenerative	O
disease	O
.	O

Mutations	O
in	O
the	O
genes	O
encoding	O
the	O
three	O
other	O
succinate	B-Gene_or_gene_product
dehydrogenase	I-Gene_or_gene_product
subunits	O
(	O
SDHB	B-Gene_or_gene_product
,	O
SDHC	B-Gene_or_gene_product
and	O
SDHD	B-Gene_or_gene_product
)	O
have	O
been	O
identified	O
in	O
patients	B-Organism
affected	O
by	O
familial	O
or	O
'	O
apparently	O
sporadic	O
'	O
paraganglioma	B-Cancer
and	O
/	O
or	O
pheochromocytoma	B-Cancer
,	O
an	O
autosomal	O
inherited	O
cancer	B-Cancer
-	O
susceptibility	O
syndrome	O
.	O

These	O
discoveries	O
have	O
dramatically	O
changed	O
the	O
work	O
-	O
up	O
and	O
genetic	O
counseling	O
of	O
patients	B-Organism
and	O
families	O
with	O
paragangliomas	B-Cancer
and	O
/	O
or	O
pheochromocytomas	B-Cancer
.	O

The	O
subsequent	O
identification	O
of	O
germline	O
mutations	O
in	O
the	O
gene	O
encoding	O
fumarase	B-Gene_or_gene_product
-	O
-	O
another	O
TCA	B-Simple_chemical
cycle	O
enzyme	O
-	O
-	O
in	O
a	O
new	O
hereditary	O
form	O
of	O
susceptibility	O
to	O
renal	B-Cancer
,	O
uterine	B-Cancer
and	O
cutaneous	B-Cancer
tumors	I-Cancer
has	O
highlighted	O
the	O
potential	O
role	O
of	O
the	O
TCA	B-Simple_chemical
cycle	O
and	O
,	O
more	O
generally	O
,	O
of	O
the	O
mitochondria	B-Cellular_component
in	O
cancer	B-Cancer
.	O

Mouse	B-Organism
p10	B-Gene_or_gene_product
,	O
an	O
alternative	O
spliced	O
form	O
of	O
p15INK4b	B-Gene_or_gene_product
,	O
inhibits	O
cell	B-Cell
cycle	O
progression	O
and	O
malignant	O
transformation	O
.	O

The	O
INK4	B-Gene_or_gene_product
family	O
of	O
proteins	O
negatively	O
regulates	O
cell	B-Cell
cycle	O
progression	O
at	O
the	O
G	O
(	O
1	O
)	O
-	O
S	O
transition	O
by	O
inhibiting	O
cyclin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
dependent	I-Gene_or_gene_product
kinases	I-Gene_or_gene_product
.	O

Two	O
of	O
these	O
cell	B-Cell
cycle	O
inhibitors	O
,	O
p16	B-Gene_or_gene_product
(	O
INK4A	B-Gene_or_gene_product
)	O
and	O
p15	B-Gene_or_gene_product
(	O
INK4B	B-Gene_or_gene_product
)	O
,	O
have	O
tumor	B-Cancer
suppressor	O
activities	O
and	O
are	O
inactivated	O
in	O
human	B-Organism
cancer	B-Cancer
.	O

Interestingly	O
,	O
both	O
INK4	B-Gene_or_gene_product
genes	O
express	O
alternative	O
splicing	O
variants	O
.	O

In	O
addition	O
to	O
p16	B-Gene_or_gene_product
(	O
INK4A	B-Gene_or_gene_product
)	O
,	O
the	O
INK4A	B-Gene_or_gene_product
locus	O
encodes	O
a	O
splice	O
variant	O
,	O
termed	O
p12	B-Gene_or_gene_product
-	O
-	O
specifically	O
expressed	O
in	O
human	B-Organism
pancreas	B-Organ
-	O
-	O
and	O
ARF	B-Gene_or_gene_product
,	O
a	O
protein	O
encoded	O
by	O
an	O
alternative	O
reading	O
frame	O
that	O
acts	O
as	O
a	O
tumor	B-Cancer
suppressor	O
through	O
the	O
p53	B-Gene_or_gene_product
pathway	O
.	O

Similarly	O
,	O
the	O
human	B-Organism
INK4B	B-Gene_or_gene_product
locus	O
encodes	O
the	O
p15	B-Gene_or_gene_product
(	O
INK4B	B-Gene_or_gene_product
)	O
tumor	B-Cancer
suppressor	O
and	O
one	O
alternatively	O
spliced	O
form	O
,	O
termed	O
as	O
p10	B-Gene_or_gene_product
.	O

We	O
show	O
here	O
that	O
p10	B-Gene_or_gene_product
,	O
which	O
arises	O
from	O
the	O
use	O
of	O
an	O
alternative	O
splice	O
donor	O
site	O
within	O
intron	B-Cellular_component
1	I-Cellular_component
,	O
is	O
conserved	O
in	O
the	O
mouse	B-Organism
genome	O
and	O
is	O
widely	O
expressed	O
in	O
mouse	B-Organism
tissues	B-Tissue
.	O

Similarly	O
to	O
mouse	B-Organism
p15	B-Gene_or_gene_product
(	O
INK4B	B-Gene_or_gene_product
)	O
,	O
p10	B-Gene_or_gene_product
expression	O
is	O
also	O
induced	O
by	O
oncogenic	O
insults	O
and	O
transforming	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
treatment	O
and	O
acts	O
as	O
a	O
cell	B-Cell
cycle	O
inhibitor	O
.	O

Importantly	O
,	O
we	O
show	O
that	O
mouse	B-Organism
p10	B-Gene_or_gene_product
is	O
able	O
to	O
induce	O
cell	B-Cell
cycle	O
arrest	O
in	O
a	O
p53	B-Gene_or_gene_product
-	O
dependent	O
manner	O
.	O

We	O
also	O
show	O
that	O
mouse	B-Organism
p10	B-Gene_or_gene_product
is	O
able	O
to	O
inhibit	O
foci	B-Cell
formation	O
and	O
anchorage	O
-	O
independent	O
growth	O
in	O
wild	O
-	O
type	O
mouse	B-Organism
embryonic	B-Cell
fibroblasts	I-Cell
,	O
and	O
that	O
these	O
antitransforming	O
properties	O
of	O
mouse	B-Organism
p10	B-Gene_or_gene_product
are	O
also	O
p53	B-Gene_or_gene_product
-	O
dependent	O
.	O

These	O
results	O
indicate	O
that	O
the	O
INK4B	B-Gene_or_gene_product
locus	O
,	O
similarly	O
to	O
INK4A	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ARF	I-Gene_or_gene_product
,	O
harbors	O
two	O
different	O
splicing	O
variants	O
that	O
can	O
be	O
involved	O
in	O
the	O
regulation	O
of	O
both	O
the	O
p53	B-Gene_or_gene_product
and	O
retinoblastoma	B-Gene_or_gene_product
pathways	O
,	O
the	O
two	O
major	O
molecular	O
pathways	O
in	O
tumor	B-Cancer
suppression	O
.	O

Rofecoxib	B-Simple_chemical
as	O
adjunctive	O
therapy	O
for	O
haemophilic	O
arthropathy	O
.	O

Joint	B-Multi-tissue_structure
haemorrhage	O
and	O
subsequent	O
haemophilic	O
arthropathy	O
are	O
significant	O
complications	O
in	O
haemophilia	O
.	O

The	O
pathophysiology	O
involves	O
inflammation	O
and	O
angiogenesis	O
.	O

Cyclooxygenase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
(	O
COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
)	O
inhibitors	O
are	O
anti	O
-	O
inflammatory	O
agents	O
,	O
which	O
have	O
potent	O
anti	O
-	O
inflammatory	O
,	O
anti	O
-	O
angiogenic	O
and	O
analgesic	O
properties	O
yet	O
do	O
not	O
affect	O
platelet	B-Cell
function	O
in	O
the	O
manner	O
of	O
traditional	O
non	B-Simple_chemical
-	I-Simple_chemical
steroidal	I-Simple_chemical
anti	I-Simple_chemical
-	I-Simple_chemical
inflammatory	I-Simple_chemical
drugs	I-Simple_chemical
.	O

These	O
properties	O
make	O
such	O
agents	O
potentially	O
useful	O
as	O
adjunctive	O
therapy	O
in	O
haemophilia	O
.	O

There	O
is	O
only	O
one	O
prior	O
report	O
describing	O
rofecoxib	B-Simple_chemical
treatment	O
in	O
a	O
single	O
haemophilia	O
patient	B-Organism
.	O

Our	O
objectives	O
were	O
to	O
determine	O
the	O
safety	O
and	O
efficacy	O
of	O
rofecoxib	B-Simple_chemical
in	O
treating	O
acute	O
haemarthrosis	O
,	O
chronic	O
synovitis	O
,	O
target	O
joints	B-Multi-tissue_structure
and	O
pain	O
.	O

We	O
conducted	O
a	O
retrospective	O
medical	O
record	O
review	O
of	O
patients	B-Organism
treated	O
with	O
rofecoxib	B-Simple_chemical
for	O
acute	O
haemarthrosis	O
,	O
chronic	O
synovitis	O
,	O
target	O
joint	B-Multi-tissue_structure
or	O
pain	O
.	O

The	O
safety	O
and	O
efficacy	O
of	O
rofecoxib	B-Simple_chemical
treatment	O
were	O
determined	O
based	O
on	O
subjective	O
patient	B-Organism
reports	O
and	O
physical	O
examinations	O
during	O
follow	O
-	O
up	O
clinic	O
visits	O
.	O

A	O
total	O
of	O
28	O
patients	B-Organism
between	O
3	O
and	O
37	O
years	O
of	O
age	O
were	O
treated	O
for	O
a	O
total	O
of	O
42	O
courses	O
of	O
rofecoxib	B-Simple_chemical
treatment	O
.	O

All	O
courses	O
were	O
evaluated	O
for	O
safety	O
and	O
31	O
for	O
efficacy	O
.	O

Rofecoxib	B-Simple_chemical
was	O
used	O
for	O
eight	O
acute	O
haemarthrosis	O
,	O
four	O
target	O
joints	B-Multi-tissue_structure
,	O
seven	O
cases	O
of	O
synovitis	O
and	O
12	O
episodes	O
of	O
pain	O
.	O

Efficacy	O
was	O
demonstrated	O
particularly	O
for	O
chronic	O
synovitis	O
and	O
pain	O
and	O
no	O
serious	O
adverse	O
events	O
occurred	O
.	O

This	O
is	O
the	O
largest	O
study	O
to	O
date	O
evaluating	O
COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
inhibitors	O
as	O
adjunctive	O
therapy	O
in	O
haemophilia	O
and	O
suggests	O
that	O
these	O
agents	O
may	O
be	O
an	O
important	O
adjunctive	O
therapy	O
in	O
the	O
management	O
of	O
haemophilia	O
.	O

Thalidomide	B-Simple_chemical
and	O
angiostatin	B-Gene_or_gene_product
inhibit	O
tumor	B-Cancer
growth	O
in	O
a	O
murine	B-Organism
xenograft	B-Cancer
model	O
of	O
human	B-Organism
cervical	B-Cancer
cancer	I-Cancer
.	O

OBJECTIVE	O
:	O
To	O
determine	O
the	O
impact	O
of	O
thalidomide	B-Simple_chemical
and	O
angiostatin	B-Gene_or_gene_product
on	O
tumor	B-Cancer
growth	O
,	O
angiogenesis	O
,	O
and	O
apoptosis	O
in	O
a	O
xenograft	B-Cancer
model	O
of	O
cervical	B-Cancer
cancer	I-Cancer
.	O

METHODS	O
:	O
Human	B-Organism
umbilical	O
endothelial	B-Cell
cells	I-Cell
were	O
treated	O
with	O
angiostatin	B-Gene_or_gene_product
or	O
thalidomide	B-Simple_chemical
and	O
bFGF	B-Gene_or_gene_product
-	O
induced	O
proliferation	O
was	O
assessed	O
with	O
the	O
MTT	O
assay	O
.	O

Human	B-Organism
cervical	B-Cell
cancer	I-Cell
cells	I-Cell
(	O
CaSki	B-Cell
and	O
SiHa	B-Cell
)	O
were	O
injected	O
into	O
the	O
flanks	B-Organism_subdivision
of	O
nude	B-Organism
mice	I-Organism
.	O

After	O
tumors	B-Cancer
developed	O
,	O
mice	B-Organism
were	O
treated	O
with	O
angiostatin	B-Gene_or_gene_product
20	O
mg	O
/	O
kg	O
/	O
day	O
or	O
thalidomide	B-Simple_chemical
200	O
mg	O
/	O
kg	O
/	O
day	O
for	O
30	O
days	O
.	O

Fractional	O
tumor	B-Cancer
growth	O
was	O
determined	O
and	O
immunohistochemical	O
analysis	O
of	O
tumors	B-Cancer
was	O
used	O
to	O
determine	O
degree	O
of	O
angiogenesis	O
.	O

TUNEL	O
assay	O
was	O
used	O
to	O
assess	O
apoptosis	O
.	O

RESULTS	O
:	O
Angiostatin	B-Gene_or_gene_product
inhibited	O
endothelial	B-Cell
cell	I-Cell
proliferation	O
by	O
50	O
-	O
60	O
%	O
.	O

Thalidomide	B-Simple_chemical
had	O
no	O
direct	O
effect	O
on	O
endothelial	B-Cell
cells	I-Cell
.	O

Angiostatin	B-Gene_or_gene_product
and	O
thalidomide	B-Simple_chemical
both	O
inhibited	O
tumor	B-Cancer
growth	O
by	O
about	O
55	O
%	O
.	O

We	O
found	O
no	O
additive	O
or	O
synergistic	O
effect	O
when	O
the	O
two	O
agents	O
were	O
combined	O
.	O

Both	O
agents	O
inhibited	O
angiogenesis	O
and	O
induced	O
apoptosis	O
when	O
compared	O
to	O
tumors	B-Cancer
from	O
control	O
animals	O
.	O

CONCLUSIONS	O
:	O
Angiostatin	B-Gene_or_gene_product
and	O
thalidomide	B-Simple_chemical
inhibit	O
tumor	B-Cancer
growth	O
,	O
angiogenesis	O
,	O
and	O
induce	O
apoptosis	O
in	O
this	O
xenograft	B-Cancer
model	O
of	O
cervical	B-Cancer
cancer	I-Cancer
.	O

Therapeutic	O
Electromagnetic	O
Field	O
(	O
TEMF	O
)	O
and	O
gamma	O
irradiation	O
on	O
human	B-Organism
breast	B-Cancer
cancer	I-Cancer
xenograft	I-Cancer
growth	O
,	O
angiogenesis	O
and	O
metastasis	O
.	O

BACKGROUND	O
:	O
The	O
effects	O
of	O
a	O
rectified	O
semi	O
-	O
sinewave	O
signal	O
(	O
15	O
mT	O
amplitude	O
,	O
120	O
pulses	O
per	O
second	O
,	O
EMF	O
Therapeutics	O
,	O
Inc	O
.	O
)	O
(	O
TEMF	O
)	O
alone	O
and	O
in	O
combination	O
with	O
gamma	O
irradiation	O
(	O
IR	O
)	O
therapy	O
in	O
nude	B-Organism
mice	I-Organism
bearing	O
a	O
human	B-Organism
MDA	B-Cancer
MB231	I-Cancer
breast	I-Cancer
cancer	I-Cancer
xenograft	I-Cancer
were	O
tested	O
.	O

Green	B-Gene_or_gene_product
fluorescence	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
transfected	O
cancer	B-Cell
cells	I-Cell
were	O
injected	O
into	O
the	O
mammary	B-Tissue
fat	I-Tissue
pad	I-Tissue
of	O
young	O
female	O
mice	B-Organism
.	O

Six	O
weeks	O
later	O
,	O
mice	B-Organism
were	O
randomly	O
divided	O
into	O
four	O
treatment	O
groups	O
:	O
untreated	O
controls	O
;	O
10	O
minute	O
daily	O
TEMF	O
;	O
200	O
cGy	O
of	O
IR	O
every	O
other	O
day	O
(	O
total	O
800	O
cGy	O
)	O
;	O
IR	O
plus	O
daily	O
TEMF	O
.	O

Some	O
mice	B-Organism
in	O
each	O
group	O
were	O
euthanized	O
24	O
hours	O
after	O
the	O
end	O
of	O
IR	O
.	O

TEMF	O
treatment	O
continued	O
for	O
3	O
additional	O
weeks	O
.	O

Tumor	B-Cancer
sections	I-Cancer
were	O
stained	O
for	O
:	O
endothelial	B-Cell
cells	I-Cell
with	O
CD31	B-Gene_or_gene_product
and	O
PAS	B-Simple_chemical
or	O
hypoxia	B-Gene_or_gene_product
inducible	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
(	O
HIF	B-Gene_or_gene_product
)	O
.	O

RESULTS	O
:	O
Most	O
tumors	B-Cancer
less	O
than	O
35	O
mm3	O
were	O
white	O
but	O
tumors	B-Cancer
greater	O
than	O
35	O
mm3	O
were	O
pink	O
and	O
had	O
a	O
vascularized	O
capsule	B-Cancer
.	O

The	O
cortex	B-Cancer
within	O
100	O
microns	O
of	O
the	O
capsule	B-Cancer
had	O
little	O
vascularization	O
.	O

Blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
,	O
capillaries	B-Tissue
,	O
and	O
endothelial	B-Cellular_component
pseudopods	I-Cellular_component
were	O
found	O
at	O
greater	O
than	O
100	O
microns	O
from	O
the	O
capsule	B-Cancer
(	O
subcortex	B-Cancer
)	O
.	O

Tumors	B-Cancer
greater	O
than	O
35	O
mm3	O
treated	O
with	O
IR	O
24	O
hours	O
previously	O
or	O
with	O
TEMF	O
had	O
decreased	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
in	O
the	O
subcortex	B-Cancer
and	O
more	O
endothelial	B-Cellular_component
pseudopods	I-Cellular_component
projecting	O
into	O
hypoxic	O
,	O
HIF	B-Gene_or_gene_product
positive	O
areas	O
than	O
tumors	B-Cancer
from	O
the	O
control	O
group	O
.	O

Mice	B-Organism
that	O
received	O
either	O
IR	O
or	O
TEMF	O
had	O
significantly	O
fewer	O
lung	B-Organ
metastatic	O
sites	O
and	O
slower	O
tumor	B-Cancer
growth	O
than	O
did	O
untreated	O
mice	B-Organism
.	O

No	O
harmful	O
side	O
effects	O
were	O
attributed	O
to	O
TEMF	O
.	O

CONCLUSION	O
:	O
TEMF	O
therapy	O
provided	O
a	O
safe	O
means	O
for	O
retarding	O
tumor	B-Cancer
vascularization	O
,	O
growth	O
and	O
metastasis	O
.	O

The	O
threshold	O
level	O
of	O
adenomatous	B-Gene_or_gene_product
polyposis	I-Gene_or_gene_product
coli	I-Gene_or_gene_product
protein	O
for	O
mouse	B-Organism
intestinal	B-Organ
tumorigenesis	O
.	O

The	O
adenomatous	B-Gene_or_gene_product
polyposis	I-Gene_or_gene_product
coli	I-Gene_or_gene_product
(	O
APC	B-Gene_or_gene_product
)	O
gene	O
,	O
whose	O
mutations	O
are	O
responsible	O
for	O
familial	O
adenomatous	O
polyposis	O
,	O
is	O
a	O
major	O
negative	O
controller	O
of	O
the	O
Wnt	B-Gene_or_gene_product
/	O
beta	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
pathway	O
.	O

To	O
investigate	O
the	O
dose	O
-	O
dependent	O
effects	O
of	O
APC	B-Gene_or_gene_product
protein	O
in	O
suppressing	O
intestinal	B-Organ
tumorigenesis	O
,	O
we	O
constructed	O
mutant	O
mice	B-Organism
carrying	O
hypomorphic	O
Apc	B-Gene_or_gene_product
alleles	O
Apc	B-Gene_or_gene_product
(	I-Gene_or_gene_product
neoR	I-Gene_or_gene_product
)	I-Gene_or_gene_product
and	O
Apc	B-Gene_or_gene_product
(	I-Gene_or_gene_product
neoF	I-Gene_or_gene_product
)	I-Gene_or_gene_product
whose	O
expression	O
levels	O
were	O
reduced	O
to	O
20	O
%	O
and	O
10	O
%	O
of	O
the	O
wild	O
type	O
,	O
respectively	O
.	O

Although	O
both	O
hypomorphic	O
heterozygotes	O
developed	O
intestinal	B-Pathological_formation
polyps	I-Pathological_formation
,	O
tumor	B-Cancer
multiplicities	O
were	O
much	O
lower	O
than	O
that	O
in	O
Apc	B-Gene_or_gene_product
(	I-Gene_or_gene_product
Delta716	I-Gene_or_gene_product
)	I-Gene_or_gene_product
mice	O
,	O
heterozygotes	O
of	O
an	O
Apc	B-Gene_or_gene_product
null	O
allele	O
.	O

Like	O
in	O
Apc	B-Gene_or_gene_product
(	I-Gene_or_gene_product
Delta716	I-Gene_or_gene_product
)	I-Gene_or_gene_product
mice	O
,	O
loss	O
of	O
the	O
wild	O
-	O
type	O
Apc	B-Gene_or_gene_product
allele	O
was	O
confirmed	O
for	O
all	O
polyps	B-Pathological_formation
examined	O
in	O
the	O
Apc	B-Gene_or_gene_product
(	I-Gene_or_gene_product
neoR	I-Gene_or_gene_product
)	I-Gene_or_gene_product
and	O
Apc	B-Gene_or_gene_product
(	I-Gene_or_gene_product
neoF	I-Gene_or_gene_product
)	I-Gene_or_gene_product
mice	O
.	O

In	O
the	O
embryonic	B-Cell
stem	I-Cell
cells	I-Cell
homozygous	O
for	O
these	O
hypomorphic	O
Apc	B-Gene_or_gene_product
alleles	O
,	O
the	O
level	O
of	O
the	O
APC	B-Gene_or_gene_product
protein	O
was	O
inversely	O
correlated	O
with	O
both	O
the	O
beta	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
accumulation	O
and	O
beta	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
/	O
T	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cell	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
transcriptional	O
activity	O
.	O

These	O
results	O
suggest	O
that	O
the	O
reduced	O
APC	B-Gene_or_gene_product
protein	O
level	O
increases	O
intestinal	B-Pathological_formation
polyp	I-Pathological_formation
multiplicity	O
through	O
quantitative	O
stimulation	O
of	O
the	O
beta	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
/	O
T	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cell	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
transcription	O
.	O

We	O
further	O
estimated	O
the	O
threshold	O
of	O
APC	B-Gene_or_gene_product
protein	O
level	O
that	O
forms	O
one	O
polyp	B-Pathological_formation
per	O
mouse	B-Organism
as	O
approximately	O
15	O
%	O
of	O
the	O
wild	O
type	O
.	O

These	O
results	O
also	O
suggest	O
therapeutic	O
implications	O
concerning	O
Wnt	B-Gene_or_gene_product
signaling	O
inhibitors	O
.	O

The	O
merits	O
of	O
vascular	B-Multi-tissue_structure
targeting	O
for	O
gynecologic	O
malignancies	B-Cancer
.	O

Neovascularization	O
is	O
an	O
early	O
and	O
critical	O
step	O
in	O
tumor	B-Cancer
development	O
and	O
progression	O
.	O

Tumor	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
are	O
distinct	O
from	O
their	O
normal	O
counterparts	O
morphologically	O
as	O
well	O
as	O
at	O
a	O
molecular	O
level	O
.	O

Recent	O
studies	O
on	O
factors	O
involved	O
in	O
tumor	B-Multi-tissue_structure
vascular	I-Multi-tissue_structure
development	O
have	O
identified	O
new	O
therapeutic	O
targets	O
for	O
inhibiting	O
tumor	B-Cancer
neovascularization	O
and	O
thus	O
tumor	B-Cancer
progression	O
.	O

However	O
,	O
the	O
process	O
of	O
tumor	B-Multi-tissue_structure
blood	I-Multi-tissue_structure
vessel	I-Multi-tissue_structure
formation	O
is	O
complex	O
,	O
and	O
each	O
tumor	B-Cancer
exhibits	O
unique	O
features	O
in	O
its	O
vasculature	B-Multi-tissue_structure
.	O

An	O
understanding	O
of	O
the	O
relative	O
contribution	O
of	O
various	O
pathways	O
in	O
the	O
development	O
of	O
tumor	B-Multi-tissue_structure
vasculature	I-Multi-tissue_structure
is	O
critical	O
for	O
developing	O
effective	O
and	O
selective	O
therapeutic	O
approaches	O
.	O

Several	O
such	O
agents	O
are	O
currently	O
in	O
clinical	O
trials	O
,	O
and	O
many	O
others	O
are	O
under	O
development	O
.	O

In	O
this	O
review	O
,	O
the	O
mechanisms	O
and	O
factors	O
involved	O
in	O
tumor	B-Multi-tissue_structure
blood	I-Multi-tissue_structure
vessel	I-Multi-tissue_structure
formation	O
are	O
discussed	O
.	O

In	O
addition	O
,	O
selected	O
novel	O
classes	O
of	O
antivascular	B-Multi-tissue_structure
therapies	O
,	O
including	O
those	O
targeting	O
tumor	B-Cell
endothelial	I-Cell
cells	I-Cell
and	O
other	O
components	O
of	O
the	O
tumor	B-Multi-tissue_structure
vasculature	I-Multi-tissue_structure
,	O
are	O
summarized	O
.	O

Normal	O
and	O
transforming	O
functions	O
of	O
RUNX1	B-Gene_or_gene_product
:	O
a	O
perspective	O
.	O

Converging	O
studies	O
from	O
many	O
investigators	O
indicate	O
that	O
RUNX1	B-Gene_or_gene_product
has	O
a	O
critical	O
role	O
in	O
the	O
correct	O
maintenance	O
of	O
essential	O
cellular	B-Cell
functions	O
during	O
embryonic	B-Developing_anatomical_structure
development	O
and	O
after	O
birth	O
.	O

The	O
discovery	O
that	O
this	O
gene	O
is	O
also	O
frequently	O
mutated	O
in	O
human	B-Organism
leukemia	B-Cancer
has	O
increased	O
the	O
interest	O
in	O
the	O
role	O
that	O
RUNX1	B-Gene_or_gene_product
plays	O
in	O
both	O
normal	O
and	O
transforming	O
pathways	O
.	O

Here	O
,	O
we	O
provide	O
an	O
overview	O
of	O
the	O
many	O
roles	O
of	O
RUNX1	B-Gene_or_gene_product
in	O
hematopoietic	B-Cell
self	O
-	O
renewal	O
and	O
differentiation	O
and	O
summarize	O
the	O
information	O
that	O
is	O
currently	O
available	O
on	O
the	O
many	O
mechanisms	O
of	O
RUNX1	B-Gene_or_gene_product
deregulation	O
in	O
human	B-Organism
leukemia	B-Cancer
.	O

Domain	O
5	O
of	O
cleaved	O
high	B-Gene_or_gene_product
molecular	I-Gene_or_gene_product
weight	I-Gene_or_gene_product
kininogen	I-Gene_or_gene_product
inhibits	O
endothelial	B-Cell
cell	I-Cell
migration	O
through	O
Akt	B-Gene_or_gene_product
.	O

Domain	O
5	O
(	O
D5	O
)	O
of	O
cleaved	O
high	B-Gene_or_gene_product
molecular	I-Gene_or_gene_product
weight	I-Gene_or_gene_product
kininogen	I-Gene_or_gene_product
(	O
HKa	B-Gene_or_gene_product
)	O
inhibits	O
angiogenesis	O
in	O
vivo	O
and	O
endothelial	B-Cell
cell	I-Cell
migration	O
in	O
vitro	O
,	O
but	O
the	O
cell	B-Cell
signaling	O
pathways	O
involved	O
in	O
HKa	B-Gene_or_gene_product
and	O
D5	O
inhibition	O
of	O
endothelial	B-Cell
cell	I-Cell
migration	O
are	O
incompletely	O
delineated	O
.	O

This	O
study	O
examines	O
the	O
mechanism	O
of	O
HKa	B-Gene_or_gene_product
and	O
D5	O
inhibition	O
of	O
two	O
potent	O
stimulators	O
of	O
endothelial	B-Cell
cell	I-Cell
migration	O
,	O
sphingosine	B-Gene_or_gene_product
1	I-Gene_or_gene_product
-	I-Gene_or_gene_product
phosphate	I-Gene_or_gene_product
(	O
S1P	B-Gene_or_gene_product
)	O
and	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
,	O
that	O
act	O
through	O
the	O
P13	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
-	O
Akt	B-Gene_or_gene_product
signaling	O
pathway	O
.	O

HKa	B-Gene_or_gene_product
and	O
D5	O
inhibit	O
bovine	B-Organism
pulmonary	B-Cell
artery	I-Cell
endothelial	I-Cell
cell	I-Cell
(	O
BPAE	B-Cell
)	O
or	O
human	B-Cell
umbilical	I-Cell
vein	I-Cell
endothelial	I-Cell
cell	I-Cell
chemotaxis	O
in	O
the	O
modified	O
-	O
Boyden	O
chamber	O
in	O
response	O
toVEGF	B-Gene_or_gene_product
or	O
S1P	B-Gene_or_gene_product
.	O

The	O
inhibition	O
of	O
migration	O
by	O
HKa	B-Gene_or_gene_product
is	O
reversed	O
by	O
antibodies	O
to	O
urokinase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
type	I-Gene_or_gene_product
plasminogen	I-Gene_or_gene_product
activator	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
.	O

Both	O
HKa	B-Gene_or_gene_product
and	O
D5	O
decrease	O
the	O
speed	O
of	O
BPAE	B-Cell
cell	I-Cell
migration	O
and	O
alter	O
the	O
morphology	O
in	O
live	O
,	O
time	O
-	O
lapse	O
microscopy	O
after	O
stimulation	O
with	O
S1P	B-Gene_or_gene_product
or	O
VEGF	B-Gene_or_gene_product
.	O

HKa	B-Gene_or_gene_product
and	O
D5	O
reduce	O
the	O
localization	O
of	O
paxillin	B-Gene_or_gene_product
to	O
the	O
focal	B-Cellular_component
adhesions	I-Cellular_component
after	O
S1P	B-Gene_or_gene_product
and	O
VEGF	B-Gene_or_gene_product
stimulation	O
.	O

To	O
better	O
understand	O
the	O
intracellular	B-Immaterial_anatomical_entity
signaling	O
pathways	O
,	O
we	O
examined	O
the	O
effect	O
of	O
HKa	B-Gene_or_gene_product
on	O
the	O
phosphorylation	O
of	O
Akt	B-Gene_or_gene_product
and	O
its	O
downstream	O
effector	O
,	O
GSK	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3alpha	I-Gene_or_gene_product
HKa	B-Gene_or_gene_product
and	O
D5	O
inhibit	O
phosphorylation	O
of	O
Akt	B-Gene_or_gene_product
and	O
GSK	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3alpha	I-Gene_or_gene_product
after	O
stimulation	O
withVEGF	B-Gene_or_gene_product
and	O
S1P	B-Gene_or_gene_product
.	O

Inhibitors	O
of	O
Akt	B-Gene_or_gene_product
and	O
P13	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
,	O
the	O
upstream	O
activator	O
of	O
Akt	B-Gene_or_gene_product
,	O
block	O
endothelial	B-Cell
cell	I-Cell
migration	O
and	O
disrupt	O
paxillin	B-Gene_or_gene_product
localization	O
to	O
the	O
focal	B-Cellular_component
adhesions	I-Cellular_component
after	O
stimulation	O
with	O
VEGF	B-Gene_or_gene_product
and	O
S1P	B-Gene_or_gene_product
.	O

Therefore	O
we	O
suggest	O
that	O
HKa	B-Gene_or_gene_product
through	O
its	O
D5	O
domain	O
alters	O
P13	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
-	O
Akt	B-Gene_or_gene_product
signaling	O
to	O
inhibit	O
endothelial	B-Cell
cell	I-Cell
migration	O
through	O
alterations	O
in	O
the	O
focal	B-Cellular_component
adhesions	I-Cellular_component
.	O

Effect	O
of	O
thalidomide	B-Simple_chemical
affecting	O
VEGF	B-Gene_or_gene_product
secretion	O
,	O
cell	B-Cell
migration	O
,	O
adhesion	O
and	O
capillary	B-Tissue
tube	I-Tissue
formation	O
of	O
human	B-Organism
endothelial	B-Cell
EA	I-Cell
.	I-Cell
hy	I-Cell
926	I-Cell
cells	I-Cell
.	O

Angiogenesis	O
,	O
new	O
blood	B-Multi-tissue_structure
vessel	I-Multi-tissue_structure
formation	O
,	O
is	O
a	O
multistep	O
process	O
,	O
precisely	O
regulated	O
by	O
pro	O
-	O
angiogenic	O
cytokines	O
,	O
which	O
stimulate	O
endothelial	B-Cell
cells	I-Cell
to	O
migrate	O
,	O
proliferate	O
and	O
differentiate	O
to	O
form	O
new	O
capillary	B-Tissue
microvessels	I-Tissue
.	O

Excessive	O
vascular	B-Multi-tissue_structure
development	O
and	O
blood	B-Multi-tissue_structure
vessel	I-Multi-tissue_structure
remodeling	O
appears	O
in	O
psoriasis	O
,	O
rheumatoid	O
arthritis	O
,	O
diabetic	O
retinopathy	O
and	O
solid	B-Cancer
tumors	I-Cancer
formation	O
.	O

Thalidomide	B-Simple_chemical
[	O
alpha	B-Simple_chemical
-	I-Simple_chemical
(	I-Simple_chemical
N	I-Simple_chemical
-	I-Simple_chemical
phthalimido	I-Simple_chemical
)	I-Simple_chemical
-	I-Simple_chemical
glutarimide	I-Simple_chemical
]	O
is	O
known	O
to	O
be	O
a	O
potent	O
inhibitor	O
of	O
angiogenesis	O
,	O
but	O
the	O
mechanism	O
of	O
its	O
inhibitory	O
action	O
remains	O
unclear	O
.	O

The	O
aim	O
of	O
the	O
study	O
was	O
to	O
investigate	O
the	O
potential	O
influence	O
of	O
thalidomide	B-Simple_chemical
on	O
the	O
several	O
steps	O
of	O
angiogenesis	O
,	O
using	O
in	O
vitro	O
models	O
.	O

We	O
have	O
evaluated	O
the	O
effect	O
of	O
thalidomide	B-Simple_chemical
on	O
VEGF	B-Gene_or_gene_product
secretion	O
,	O
cell	B-Cell
migration	O
,	O
adhesion	O
as	O
well	O
as	O
in	O
capillary	B-Tissue
formation	O
of	O
human	B-Organism
endothelial	B-Cell
cell	I-Cell
line	I-Cell
EA	I-Cell
.	I-Cell
hy	I-Cell
926	I-Cell
.	O

Thalidomide	B-Simple_chemical
at	O
the	O
concentrations	O
of	O
0	O
.	O
01	O
microM	O
and	O
10	O
microM	O
inhibited	O
VEGF	B-Gene_or_gene_product
secretion	O
into	O
supernatants	O
,	O
decreased	O
the	O
number	O
of	O
formed	O
capillary	B-Tissue
tubes	I-Tissue
and	O
increased	O
cell	B-Cell
adhesion	O
to	O
collagen	B-Gene_or_gene_product
.	O

Administration	O
of	O
thalidomide	B-Simple_chemical
at	O
the	O
concentration	O
of	O
0	O
.	O
01	O
microM	O
increased	O
cell	B-Cell
migration	O
,	O
while	O
at	O
10	O
microM	O
,	O
it	O
decreased	O
cell	B-Cell
migration	O
.	O

Thalidomide	B-Simple_chemical
in	O
concentrations	O
from	O
0	O
.	O
1	O
microM	O
to	O
10	O
microM	O
did	O
not	O
change	O
cell	B-Cell
proliferation	O
of	O
72	O
-	O
h	O
cell	B-Cell
cultures	I-Cell
.	O

We	O
conclude	O
that	O
anti	O
-	O
angiogenic	O
action	O
of	O
thalidomide	B-Simple_chemical
is	O
due	O
to	O
direct	O
inhibitory	O
action	O
on	O
VEGF	B-Gene_or_gene_product
secretion	O
and	O
capillary	B-Tissue
microvessel	I-Tissue
formation	O
as	O
well	O
as	O
immunomodulatory	O
influence	O
on	O
EA	B-Cell
.	I-Cell
hy	I-Cell
926	I-Cell
cells	I-Cell
migration	O
and	O
adhesion	O
.	O

Regulation	O
of	O
skin	B-Tissue
microvasculature	I-Tissue
angiogenesis	O
,	O
cell	B-Cell
migration	O
,	O
and	O
permeability	O
by	O
a	O
specific	O
inhibitor	O
of	O
PKCalpha	B-Gene_or_gene_product
.	O

Activation	O
of	O
protein	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
C	I-Gene_or_gene_product
(	O
PKC	B-Gene_or_gene_product
)	O
induces	O
phenotypic	O
changes	O
in	O
the	O
morphology	O
of	O
microvascular	B-Cell
endothelial	I-Cell
cells	I-Cell
that	O
affect	O
major	O
functions	O
of	O
the	O
microvasculature	B-Tissue
.	O

These	O
functions	O
include	O
the	O
first	O
stages	O
of	O
sprouting	O
in	O
angiogenesis	O
,	O
cell	B-Cell
migration	O
following	O
wounding	O
,	O
and	O
vascular	B-Multi-tissue_structure
permeability	O
.	O

The	O
specific	O
isoform	O
(	O
s	O
)	O
of	O
PKC	B-Gene_or_gene_product
responsible	O
for	O
each	O
of	O
these	O
changes	O
has	O
not	O
been	O
previously	O
identified	O
.	O

In	O
this	O
study	O
,	O
we	O
used	O
two	O
inflammatory	O
agents	O
,	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1beta	I-Gene_or_gene_product
and	O
phorbol	B-Simple_chemical
myristic	I-Simple_chemical
acetate	I-Simple_chemical
,	O
to	O
activate	O
PKC	B-Gene_or_gene_product
isozymes	O
and	O
specific	O
inhibitors	O
of	O
PKCalpha	B-Gene_or_gene_product
(	O
Go6976	B-Simple_chemical
)	O
and	O
PKCbeta	B-Gene_or_gene_product
(	O
hispidin	B-Simple_chemical
)	O
to	O
distinguish	O
how	O
each	O
of	O
these	O
isoform	O
(	O
s	O
)	O
controls	O
angiogenesis	O
,	O
wound	B-Pathological_formation
healing	O
,	O
and	O
permeability	O
.	O

In	O
all	O
cases	O
,	O
only	O
inhibition	O
of	O
PKCalpha	B-Gene_or_gene_product
inhibited	O
each	O
of	O
these	O
functions	O
when	O
compared	O
to	O
the	O
inhibition	O
of	O
PKCbeta	B-Gene_or_gene_product
.	O

Additional	O
analysis	O
of	O
the	O
mechanism	O
of	O
action	O
of	O
Go6976	B-Simple_chemical
(	O
RT	O
-	O
PCR	O
,	O
Western	O
blots	O
,	O
and	O
immunohistochemistry	O
)	O
of	O
the	O
changes	O
in	O
the	O
phosphorylated	O
and	O
nonphosphorylated	O
forms	O
of	O
PKCalpha	B-Gene_or_gene_product
in	O
the	O
cell	B-Cellular_component
membrane	I-Cellular_component
and	O
cytoplasm	B-Organism_substance
confirmed	O
the	O
specificity	O
of	O
PKCalpha	B-Gene_or_gene_product
inhibition	O
by	O
Go6976	B-Simple_chemical
.	O

These	O
studies	O
therefore	O
indicate	O
a	O
specific	O
and	O
a	O
regulatory	O
role	O
of	O
the	O
PKCalpha	B-Gene_or_gene_product
isoform	O
in	O
three	O
major	O
endothelial	B-Cell
cell	I-Cell
functions	O
that	O
are	O
important	O
in	O
the	O
maintenance	O
of	O
microvascular	B-Tissue
homeostasis	O
.	O

The	O
differential	O
regulation	O
of	O
human	B-Organism
telomerase	B-Gene_or_gene_product
reverse	I-Gene_or_gene_product
transcriptase	I-Gene_or_gene_product
and	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
may	O
contribute	O
to	O
the	O
clinically	O
more	O
aggressive	O
behavior	O
of	O
p63	B-Gene_or_gene_product
-	O
positive	O
breast	O
carcinomas	O
.	O

p63	B-Gene_or_gene_product
,	O
a	O
p53	B-Gene_or_gene_product
homologue	O
,	O
is	O
a	O
myoepithelial	B-Cell
cell	I-Cell
marker	O
in	O
the	O
normal	O
mammary	B-Organ
gland	I-Organ
but	O
p63	B-Gene_or_gene_product
-	O
positive	O
neoplastic	O
cells	O
may	O
be	O
found	O
in	O
up	O
to	O
11	O
%	O
of	O
invasive	B-Cancer
breast	I-Cancer
carcinomas	I-Cancer
.	O

This	O
study	O
aims	O
to	O
verify	O
the	O
relationship	O
between	O
p63	B-Gene_or_gene_product
expression	O
and	O
several	O
clinicopathological	O
features	O
and	O
tumor	B-Cancer
markers	O
of	O
clinical	O
significance	O
in	O
breast	B-Cancer
pathology	I-Cancer
including	O
key	O
regulators	O
of	O
the	O
cell	B-Cell
cycle	O
,	O
oncogenes	O
,	O
apoptosis	O
-	O
related	O
proteins	O
,	O
metalloproteinases	B-Gene_or_gene_product
and	O
their	O
inhibitors	O
.	O

Immunohistochemistry	O
with	O
27	O
primary	O
antibodies	O
was	O
performed	O
in	O
100	O
formalin	O
-	O
fixed	O
paraffin	O
-	O
embedded	O
samples	O
of	O
invasive	B-Cancer
ductal	I-Cancer
carcinomas	I-Cancer
.	O

p63	B-Gene_or_gene_product
-	O
positive	O
cells	O
were	O
found	O
in	O
16	O
%	O
of	O
carcinomas	B-Cancer
.	O

p63	B-Gene_or_gene_product
-	O
positive	O
carcinomas	O
were	O
poorly	O
differentiated	O
,	O
hormone	O
receptor	O
-	O
negative	O
neoplasms	B-Cancer
with	O
a	O
high	O
proliferation	O
rate	O
.	O

p63	B-Gene_or_gene_product
also	O
correlated	O
with	O
advanced	O
pathological	O
stage	O
,	O
tumor	B-Cancer
size	O
,	O
and	O
the	O
expression	O
of	O
human	B-Organism
telomerase	B-Gene_or_gene_product
reverse	I-Gene_or_gene_product
transcriptase	I-Gene_or_gene_product
(	O
hTERT	B-Gene_or_gene_product
)	O
,	O
tissue	B-Gene_or_gene_product
inhibitor	I-Gene_or_gene_product
of	I-Gene_or_gene_product
matrix	I-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
TIMP1	B-Gene_or_gene_product
)	O
and	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
.	O

The	O
expression	O
of	O
TIMP1	B-Gene_or_gene_product
suggests	O
that	O
the	O
anti	O
-	O
proteolytic	O
stimuli	O
may	O
be	O
preponderant	O
in	O
p63	B-Gene_or_gene_product
-	O
positive	O
carcinomas	O
.	O

hTERT	B-Gene_or_gene_product
activity	O
is	O
associated	O
with	O
nodal	B-Multi-tissue_structure
metastases	O
and	O
cellular	B-Cell
proliferation	O
.	O

VEGF	B-Gene_or_gene_product
regulates	O
angiogenesis	O
,	O
which	O
is	O
also	O
a	O
fundamental	O
event	O
in	O
the	O
process	O
of	O
tumor	B-Cancer
growth	O
and	O
metastatic	O
dissemination	O
.	O

Thus	O
,	O
the	O
differential	O
regulation	O
of	O
hTERT	B-Gene_or_gene_product
and	O
VEGF	B-Gene_or_gene_product
in	O
p63	B-Gene_or_gene_product
-	O
positive	O
breast	O
carcinomas	O
may	O
contribute	O
to	O
the	O
clinically	O
more	O
aggressive	O
behavior	O
of	O
these	O
neoplasms	B-Cancer
.	O

Regulation	O
of	O
tumor	B-Cancer
angiogenesis	O
by	O
thrombospondin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

Angiogenesis	O
plays	O
a	O
critical	O
role	O
in	O
the	O
growth	O
and	O
metastasis	O
of	O
tumors	B-Cancer
.	O

Thrombospondin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
is	O
a	O
potent	O
angiogenesis	O
inhibitor	O
,	O
and	O
down	O
-	O
regulation	O
of	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
has	O
been	O
suggested	O
to	O
alter	O
tumor	B-Cancer
growth	O
by	O
modulating	O
angiogenesis	O
in	O
a	O
variety	O
of	O
tumor	B-Cancer
types	O
.	O

Expression	O
of	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
is	O
up	O
-	O
regulated	O
by	O
the	O
tumor	B-Cancer
suppressor	O
gene	O
,	O
p53	B-Gene_or_gene_product
,	O
and	O
down	O
-	O
regulated	O
by	O
oncogenes	O
such	O
as	O
Myc	B-Gene_or_gene_product
and	O
Ras	B-Gene_or_gene_product
.	O

TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
inhibits	O
angiogenesis	O
by	O
inhibiting	O
endothelial	B-Cell
cell	I-Cell
migration	O
and	O
proliferation	O
and	O
by	O
inducing	O
apoptosis	O
.	O

In	O
addition	O
,	O
activation	O
of	O
transforming	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
(	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
)	O
by	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
plays	O
a	O
crucial	O
role	O
in	O
the	O
regulation	O
of	O
tumor	B-Cancer
progression	O
.	O

An	O
understanding	O
of	O
the	O
molecular	O
basis	O
of	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
mediated	O
inhibition	O
of	O
angiogenesis	O
and	O
tumor	B-Cancer
progression	O
will	O
aid	O
in	O
the	O
development	O
of	O
novel	O
therapeutics	O
for	O
the	O
treatment	O
of	O
cancer	B-Cancer
.	O

Regulation	O
of	O
the	O
composition	O
of	O
the	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
by	O
low	B-Gene_or_gene_product
density	I-Gene_or_gene_product
lipoprotein	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
related	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
:	O
activities	O
based	O
on	O
regulation	O
of	O
mRNA	O
expression	O
.	O

Low	B-Gene_or_gene_product
density	I-Gene_or_gene_product
lipoprotein	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
related	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
LRP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
is	O
a	O
catabolic	O
receptor	O
for	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
(	O
ECM	B-Cellular_component
)	O
structural	O
proteins	O
and	O
for	O
proteins	O
that	O
bind	O
to	O
ECM	B-Cellular_component
.	O

LRP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
also	O
is	O
implicated	O
in	O
integrin	B-Gene_or_gene_product
maturation	O
.	O

In	O
this	O
study	O
,	O
we	O
applied	O
a	O
proteomics	O
strategy	O
to	O
identify	O
novel	O
proteins	O
involved	O
in	O
ECM	B-Cellular_component
modeling	O
that	O
are	O
regulated	O
by	O
LRP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

We	O
show	O
that	O
LRP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
deficiency	O
in	O
murine	B-Organism
embryonic	B-Cell
fibroblasts	I-Cell
(	O
MEFs	B-Cell
)	O
is	O
associated	O
with	O
increased	O
levels	O
of	O
type	B-Gene_or_gene_product
III	I-Gene_or_gene_product
collagen	I-Gene_or_gene_product
and	O
pigment	B-Gene_or_gene_product
epithelium	I-Gene_or_gene_product
-	I-Gene_or_gene_product
derived	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
,	O
which	O
accumulate	O
in	O
the	O
substratum	B-Cellular_component
surrounding	O
cells	B-Cell
.	O

The	O
collagen	B-Gene_or_gene_product
receptor	O
,	O
uPAR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
AP	I-Gene_or_gene_product
/	O
Endo	B-Gene_or_gene_product
-	I-Gene_or_gene_product
180	I-Gene_or_gene_product
,	O
is	O
also	O
increased	O
in	O
LRP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
deficient	O
MEFs	B-Cell
.	O

Human	B-Organism
LRP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
reversed	O
the	O
changes	O
in	O
protein	O
expression	O
associated	O
with	O
LRP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
deficiency	O
;	O
however	O
,	O
the	O
endocytic	O
activity	O
of	O
LRP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
was	O
not	O
involved	O
.	O

Instead	O
,	O
regulation	O
occurred	O
at	O
the	O
mRNA	O
level	O
.	O

Inhibition	O
of	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Jun	I-Gene_or_gene_product
amino	I-Gene_or_gene_product
-	I-Gene_or_gene_product
terminal	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
(	O
JNK	B-Gene_or_gene_product
)	O
blocked	O
type	B-Gene_or_gene_product
III	I-Gene_or_gene_product
collagen	I-Gene_or_gene_product
expression	O
in	O
LRP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
deficient	O
MEFs	B-Cell
,	O
suggesting	O
regulation	O
of	O
JNK	B-Gene_or_gene_product
activity	O
as	O
a	O
mechanism	O
by	O
which	O
LRP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
controls	O
mRNA	O
expression	O
.	O

The	O
ability	O
of	O
LRP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
to	O
regulate	O
expression	O
of	O
the	O
factors	O
identified	O
here	O
suggests	O
a	O
role	O
for	O
LRP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
in	O
determining	O
blood	B-Multi-tissue_structure
vessel	I-Multi-tissue_structure
structure	O
and	O
in	O
angiogenesis	O
.	O

L	B-Simple_chemical
-	I-Simple_chemical
Carnitine	I-Simple_chemical
ameliorates	O
methotrexate	B-Simple_chemical
-	O
induced	O
oxidative	O
organ	B-Organ
injury	O
and	O
inhibits	O
leukocyte	B-Cell
death	O
.	O

Methotrexate	B-Simple_chemical
(	O
MTX	B-Simple_chemical
)	O
,	O
a	O
folic	B-Simple_chemical
acid	I-Simple_chemical
antagonist	O
widely	O
used	O
for	O
the	O
treatment	O
of	O
a	O
variety	O
of	O
tumors	B-Cancer
and	O
inflammatory	O
diseases	O
,	O
affects	O
normal	B-Tissue
tissues	I-Tissue
that	O
have	O
a	O
high	O
rate	O
of	O
proliferation	O
,	O
including	O
the	O
hematopoietic	B-Cell
cells	I-Cell
of	O
the	O
bone	B-Multi-tissue_structure
marrow	I-Multi-tissue_structure
and	O
the	O
gastrointestinal	B-Cell
mucosal	I-Cell
cells	I-Cell
.	O

To	O
elucidate	O
the	O
role	O
of	O
free	O
radicals	O
and	O
leukocytes	B-Cell
in	O
MTX	B-Simple_chemical
-	O
induced	O
oxidative	O
organ	B-Organ
damage	O
and	O
the	O
putative	O
protective	O
effect	O
of	O
L	B-Simple_chemical
-	I-Simple_chemical
carnitine	I-Simple_chemical
(	O
L	B-Simple_chemical
-	I-Simple_chemical
Car	I-Simple_chemical
)	O
,	O
Wistar	B-Organism
albino	I-Organism
rats	I-Organism
were	O
administered	O
a	O
single	O
dose	O
of	O
MTX	B-Simple_chemical
(	O
20	O
mg	O
/	O
kg	O
)	O
followed	O
by	O
either	O
saline	B-Simple_chemical
or	O
L	B-Simple_chemical
-	I-Simple_chemical
Car	I-Simple_chemical
(	O
500	O
mg	O
/	O
kg	O
)	O
for	O
5	O
days	O
.	O

After	O
decapitation	O
of	O
the	O
rats	B-Organism
,	O
trunk	B-Organism_substance
blood	I-Organism_substance
was	O
obtained	O
,	O
and	O
the	O
ileum	B-Multi-tissue_structure
,	O
liver	B-Organ
,	O
and	O
kidney	B-Organ
were	O
removed	O
for	O
histological	O
examination	O
and	O
for	O
the	O
measurement	O
of	O
malondialdehyde	B-Simple_chemical
(	O
MDA	B-Simple_chemical
)	O
and	O
glutathione	B-Simple_chemical
(	O
GSH	B-Simple_chemical
)	O
levels	O
,	O
myeloperoxidase	B-Simple_chemical
(	O
MPO	B-Simple_chemical
)	O
activity	O
,	O
and	O
collagen	B-Gene_or_gene_product
content	O
.	O

Our	O
results	O
showed	O
that	O
MTX	B-Simple_chemical
administration	O
increased	O
the	O
MDA	B-Simple_chemical
and	O
MPO	B-Simple_chemical
activities	O
and	O
collagen	B-Gene_or_gene_product
content	O
and	O
decreased	O
GSH	B-Simple_chemical
levels	O
in	O
all	O
tissues	B-Tissue
,	O
while	O
these	O
alterations	O
were	O
reversed	O
in	O
L	B-Simple_chemical
-	I-Simple_chemical
Car	I-Simple_chemical
-	O
treated	O
group	O
.	O

The	O
elevated	O
serum	B-Organism_substance
TNF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
level	O
observed	O
following	O
MTX	B-Simple_chemical
treatment	O
was	O
depressed	O
with	O
L	B-Simple_chemical
-	I-Simple_chemical
Car	I-Simple_chemical
.	O

The	O
oxidative	O
burst	O
of	O
neutrophils	B-Cell
stimulated	O
by	O
Annexin	B-Gene_or_gene_product
V	I-Gene_or_gene_product
was	O
reduced	O
in	O
the	O
saline	B-Simple_chemical
-	O
treated	O
MTX	B-Simple_chemical
group	O
,	O
while	O
L	B-Simple_chemical
-	I-Simple_chemical
Car	I-Simple_chemical
abolished	O
this	O
inhibition	O
.	O

Similarly	O
,	O
flow	O
cytometric	O
measurements	O
revealed	O
that	O
leukocyte	B-Cell
apoptosis	O
was	O
increased	O
in	O
MTX	B-Simple_chemical
-	O
treated	O
animals	O
,	O
while	O
L	B-Simple_chemical
-	I-Simple_chemical
Car	I-Simple_chemical
reversed	O
these	O
effects	O
.	O

Severe	O
degeneration	O
of	O
the	O
intestinal	B-Multi-tissue_structure
mucosa	I-Multi-tissue_structure
,	O
liver	B-Multi-tissue_structure
parenchyma	I-Multi-tissue_structure
,	O
and	O
glomerular	B-Tissue
and	O
tubular	B-Tissue
epithelium	I-Tissue
observed	O
in	O
the	O
saline	B-Simple_chemical
-	O
treated	O
MTX	B-Simple_chemical
group	O
was	O
improved	O
by	O
L	B-Simple_chemical
-	I-Simple_chemical
Car	I-Simple_chemical
treatment	O
.	O

These	O
results	O
suggest	O
that	O
L	B-Simple_chemical
-	I-Simple_chemical
Car	I-Simple_chemical
,	O
possibly	O
via	O
its	O
free	O
radical	O
scavenging	O
and	O
antioxidant	O
properties	O
,	O
ameliorates	O
MTX	B-Simple_chemical
-	O
induced	O
oxidative	O
organ	B-Organ
injury	O
and	O
inhibits	O
leukocyte	B-Cell
apoptosis	O
.	O

Thus	O
,	O
supplementation	O
with	O
L	B-Simple_chemical
-	I-Simple_chemical
Carnitine	I-Simple_chemical
as	O
an	O
adjuvant	O
therapy	O
may	O
be	O
promising	O
in	O
alleviating	O
the	O
systemic	O
side	O
-	O
effects	O
of	O
chemotherapeutics	O
.	O

Pericytes	B-Cell
limit	O
tumor	B-Cell
cell	I-Cell
metastasis	O
.	O

Previously	O
we	O
observed	O
that	O
neural	B-Gene_or_gene_product
cell	I-Gene_or_gene_product
adhesion	I-Gene_or_gene_product
molecule	I-Gene_or_gene_product
(	O
NCAM	B-Gene_or_gene_product
)	O
deficiency	O
in	O
beta	B-Cell
tumor	I-Cell
cells	I-Cell
facilitates	O
metastasis	O
into	O
distant	O
organs	B-Organ
and	O
local	O
lymph	B-Multi-tissue_structure
nodes	I-Multi-tissue_structure
.	O

Here	O
,	O
we	O
show	O
that	O
NCAM	B-Gene_or_gene_product
-	O
deficient	O
beta	O
cell	O
tumors	O
grew	O
leaky	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
with	O
perturbed	O
pericyte	B-Cell
-	O
endothelial	B-Cell
cell	B-Cell
-	O
cell	B-Cell
interactions	O
and	O
deficient	O
perivascular	B-Immaterial_anatomical_entity
deposition	O
of	O
ECM	B-Cellular_component
components	I-Cellular_component
.	O

Conversely	O
,	O
tumor	B-Cell
cell	I-Cell
expression	O
of	O
NCAM	B-Gene_or_gene_product
in	O
a	O
fibrosarcoma	B-Cancer
model	O
(	O
T241	B-Cancer
)	O
improved	O
pericyte	B-Cell
recruitment	O
and	O
increased	O
perivascular	B-Immaterial_anatomical_entity
deposition	O
of	O
ECM	B-Cellular_component
molecules	I-Cellular_component
.	O

Together	O
,	O
these	O
findings	O
suggest	O
that	O
NCAM	B-Gene_or_gene_product
may	O
limit	O
tumor	B-Cell
cell	I-Cell
metastasis	O
by	O
stabilizing	O
the	O
microvessel	B-Tissue
wall	I-Tissue
.	O

To	O
directly	O
address	O
whether	O
pericyte	B-Cell
dysfunction	O
increases	O
the	O
metastatic	O
potential	O
of	O
solid	B-Cancer
tumors	I-Cancer
,	O
we	O
studied	O
beta	B-Cell
cell	I-Cell
tumorigenesis	O
in	O
primary	O
pericyte	B-Cell
-	O
deficient	O
Pdgfb	B-Gene_or_gene_product
(	O
ret	O
/	O
ret	O
)	O
mice	O
.	O

This	O
resulted	O
in	O
beta	B-Cell
tumor	I-Cell
cell	I-Cell
metastases	O
in	O
distant	O
organs	B-Organ
and	O
local	O
lymph	B-Multi-tissue_structure
nodes	I-Multi-tissue_structure
,	O
demonstrating	O
a	O
role	O
for	O
pericytes	B-Cell
in	O
limiting	O
tumor	B-Cell
cell	I-Cell
metastasis	O
.	O

These	O
data	O
support	O
a	O
new	O
model	O
for	O
how	O
tumor	B-Cell
cells	I-Cell
trigger	O
metastasis	O
by	O
perturbing	O
pericyte	B-Cell
-	O
endothelial	B-Cell
cell	B-Cell
-	O
cell	B-Cell
interactions	O
.	O

NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
and	O
IKK	B-Gene_or_gene_product
as	O
therapeutic	O
targets	O
in	O
cancer	B-Cancer
.	O

The	O
transcription	O
factor	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
and	O
associated	O
regulatory	O
factors	O
(	O
including	O
IkappaB	B-Gene_or_gene_product
kinase	O
subunits	O
and	O
the	O
IkappaB	B-Gene_or_gene_product
family	O
member	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
)	O
are	O
strongly	O
implicated	O
in	O
a	O
variety	O
of	O
hematologic	B-Cancer
and	O
solid	B-Cancer
tumor	I-Cancer
malignancies	I-Cancer
.	O

A	O
role	O
for	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
in	O
cancer	B-Cell
cells	I-Cell
appears	O
to	O
involve	O
regulation	O
of	O
cell	B-Cell
proliferation	O
,	O
control	O
of	O
apoptosis	O
,	O
promotion	O
of	O
angiogenesis	O
,	O
and	O
stimulation	O
of	O
invasion	O
/	O
metastasis	O
.	O

Consistent	O
with	O
a	O
role	O
for	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
in	O
oncogenesis	O
are	O
observations	O
that	O
inhibition	O
of	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
alone	O
or	O
in	O
combination	O
with	O
cancer	B-Cancer
therapies	O
leads	O
to	O
tumor	B-Cell
cell	I-Cell
death	O
or	O
growth	O
inhibition	O
.	O

However	O
,	O
other	O
experimental	O
data	O
indicate	O
that	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
can	O
play	O
a	O
tumor	B-Cancer
suppressor	O
role	O
in	O
certain	O
settings	O
and	O
that	O
it	O
can	O
be	O
important	O
in	O
promoting	O
an	O
apoptotic	O
signal	O
downstream	O
of	O
certain	O
cancer	B-Cancer
therapy	O
regimens	O
.	O

In	O
order	O
to	O
appropriately	O
move	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
inhibitors	O
in	O
the	O
clinic	O
,	O
thorough	O
approaches	O
must	O
be	O
initiated	O
to	O
determine	O
the	O
molecular	O
mechanisms	O
that	O
dictate	O
the	O
complexity	O
of	O
oncologic	O
and	O
therapeutic	O
outcomes	O
that	O
are	O
controlled	O
by	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
.	O

Runx2	B-Gene_or_gene_product
and	O
MYC	B-Gene_or_gene_product
collaborate	O
in	O
lymphoma	B-Cancer
development	O
by	O
suppressing	O
apoptotic	O
and	O
growth	O
arrest	O
pathways	O
in	O
vivo	O
.	O

Members	O
of	O
the	O
Runx	B-Gene_or_gene_product
and	O
MYC	B-Gene_or_gene_product
families	O
have	O
been	O
implicated	O
as	O
collaborating	O
oncogenes	O
.	O

The	O
mechanism	O
of	O
this	O
potent	O
collaboration	O
is	O
elucidated	O
in	O
this	O
study	O
of	O
Runx2	B-Gene_or_gene_product
/	O
MYC	B-Gene_or_gene_product
mice	O
.	O

As	O
shown	O
previously	O
,	O
ectopic	O
expression	O
of	O
Runx2	B-Gene_or_gene_product
in	O
the	O
thymus	B-Organ
leads	O
to	O
a	O
preneoplastic	O
state	O
defined	O
by	O
an	O
accumulation	O
of	O
cells	B-Cell
with	O
an	O
immature	O
phenotype	O
and	O
a	O
low	O
proliferative	O
rate	O
.	O

We	O
now	O
show	O
that	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
MYC	I-Gene_or_gene_product
overexpression	O
is	O
sufficient	O
to	O
rescue	O
proliferation	O
and	O
to	O
release	O
the	O
differentiation	O
block	O
imposed	O
by	O
Runx2	B-Gene_or_gene_product
.	O

Analysis	O
of	O
Runx2	B-Gene_or_gene_product
-	O
expressing	O
lymphomas	B-Cancer
reveals	O
a	O
consistently	O
low	O
rate	O
of	O
apoptosis	O
,	O
in	O
contrast	O
to	O
lymphomas	B-Cancer
of	O
MYC	B-Gene_or_gene_product
mice	O
which	O
are	O
often	O
highly	O
apoptotic	O
.	O

The	O
low	O
apoptosis	O
phenotype	O
is	O
dominant	O
in	O
Runx2	B-Gene_or_gene_product
/	O
MYC	B-Gene_or_gene_product
tumors	O
,	O
indicating	O
that	O
Runx2	B-Gene_or_gene_product
confers	O
a	O
potent	O
survival	O
advantage	O
to	O
MYC	B-Gene_or_gene_product
-	O
expressing	O
tumor	B-Cell
cells	I-Cell
.	O

The	O
role	O
of	O
the	O
p53	B-Gene_or_gene_product
pathway	O
in	O
Runx2	B-Gene_or_gene_product
/	O
MYC	B-Gene_or_gene_product
tumors	O
was	O
explored	O
on	O
a	O
p53	B-Gene_or_gene_product
heterozygote	O
background	O
.	O

Surprisingly	O
,	O
functional	O
p53	B-Gene_or_gene_product
was	O
retained	O
in	O
vivo	O
,	O
even	O
after	O
transplantation	O
,	O
whereas	O
explanted	O
tumor	B-Cell
cells	I-Cell
displayed	O
rapid	O
allele	O
loss	O
in	O
vitro	O
.	O

Our	O
results	O
show	O
that	O
Runx2	B-Gene_or_gene_product
and	O
MYC	B-Gene_or_gene_product
overcome	O
distinct	O
"	O
fail	O
-	O
safe	O
"	O
responses	O
and	O
that	O
their	O
selection	O
as	O
collaborating	O
genes	O
is	O
due	O
to	O
their	O
ability	O
to	O
neutralize	O
each	O
other	O
'	O
s	O
negative	O
growth	O
effect	O
.	O

Furthermore	O
,	O
the	O
Runx2	B-Gene_or_gene_product
/	O
MYC	B-Gene_or_gene_product
combination	O
overcomes	O
the	O
requirement	O
for	O
genetic	O
inactivation	O
of	O
the	O
p53	B-Gene_or_gene_product
pathway	O
in	O
vivo	O
.	O

Inhibition	O
of	O
the	O
mammalian	B-Gene_or_gene_product
target	I-Gene_or_gene_product
of	I-Gene_or_gene_product
rapamycin	I-Gene_or_gene_product
(	O
mTOR	B-Gene_or_gene_product
)	O
by	O
rapamycin	B-Simple_chemical
increases	O
chemosensitivity	O
of	O
CaSki	B-Cell
cells	I-Cell
to	O
paclitaxel	B-Simple_chemical
.	O

Paclitaxel	B-Simple_chemical
,	O
a	O
potent	O
anti	O
-	O
neoplastic	O
agent	O
,	O
has	O
been	O
found	O
to	O
be	O
effective	O
against	O
several	O
tumours	B-Cancer
,	O
including	O
cervical	B-Cancer
cancer	I-Cancer
.	O

However	O
,	O
the	O
exact	O
mechanism	O
underlying	O
the	O
cytotoxic	O
effects	O
of	O
pacitaxel	B-Simple_chemical
,	O
especially	O
in	O
the	O
survival	O
-	O
signalling	O
pathway	O
,	O
is	O
poorly	O
understood	O
.	O

The	O
aim	O
of	O
this	O
study	O
was	O
to	O
investigate	O
the	O
molecular	O
pathway	O
of	O
the	O
cytotoxic	O
effect	O
of	O
paclitaxel	B-Simple_chemical
in	O
human	B-Organism
cervical	B-Cell
cancer	I-Cell
cell	I-Cell
lines	I-Cell
.	O

Four	O
human	B-Organism
cervical	B-Cell
cancer	I-Cell
cell	I-Cell
lines	I-Cell
were	O
treated	O
for	O
24	O
h	O
with	O
various	O
concentration	O
of	O
paclitaxel	B-Simple_chemical
,	O
and	O
the	O
sensitivity	O
was	O
analysed	O
by	O
an	O
MTT	O
assay	O
.	O

The	O
cell	B-Cell
cycle	O
progression	O
and	O
sub	O
-	O
G1	O
population	O
were	O
analysed	O
by	O
flow	O
cytometry	O
.	O

Apoptosis	O
was	O
further	O
measured	O
by	O
DNA	B-Cellular_component
fragmentation	O
and	O
microscope	O
examination	O
.	O

The	O
protein	O
expression	O
was	O
determined	O
by	O
Western	O
blot	O
analysis	O
.	O

Our	O
results	O
showed	O
that	O
HeLa	B-Cell
cells	I-Cell
demonstrated	O
the	O
highest	O
sensitivity	O
to	O
paclitaxel	B-Simple_chemical
,	O
whereas	O
CaSki	B-Cell
cells	I-Cell
showed	O
the	O
lowest	O
.	O

In	O
cervical	B-Cell
cancer	I-Cell
cells	I-Cell
,	O
paclitaxel	B-Simple_chemical
induced	O
apoptosis	O
through	O
an	O
intrinsic	O
pathway	O
with	O
prior	O
G2	O
/	O
M	O
arrest	O
.	O

In	O
addition	O
,	O
we	O
showed	O
that	O
paclitaxel	B-Simple_chemical
downregulated	O
the	O
phosphorylation	O
of	O
Akt	B-Gene_or_gene_product
in	O
both	O
HeLa	B-Cell
and	O
CaSki	B-Cell
cells	I-Cell
.	O

Interestingly	O
,	O
in	O
CaSki	B-Cell
cells	I-Cell
,	O
which	O
were	O
more	O
suggestive	O
of	O
a	O
resistant	O
phenotype	O
,	O
paclitaxel	B-Simple_chemical
induced	O
the	O
activation	O
of	O
mTOR	B-Gene_or_gene_product
as	O
a	O
downstream	O
target	O
of	O
Akt	B-Gene_or_gene_product
.	O

Pre	O
-	O
treatment	O
with	O
rapamycin	B-Simple_chemical
inhibited	O
activation	O
of	O
mTOR	B-Gene_or_gene_product
signalling	O
and	O
significantly	O
enhanced	O
the	O
sensitivity	O
of	O
CaSki	B-Cell
cells	I-Cell
to	O
paclitaxel	B-Simple_chemical
by	O
increasing	O
apoptotic	O
cell	B-Cell
death	O
.	O

This	O
effect	O
was	O
mediated	O
,	O
at	O
least	O
partly	O
,	O
through	O
caspase	B-Gene_or_gene_product
activation	O
.	O

Overall	O
,	O
paclitaxel	B-Simple_chemical
exerts	O
its	O
anti	O
-	O
tumour	B-Cancer
effects	O
on	O
cervical	B-Cell
cancer	I-Cell
cells	I-Cell
by	O
inducing	O
apoptosis	O
through	O
intrinsic	O
pathway	O
,	O
and	O
rapamycin	B-Simple_chemical
targeted	O
to	O
mTOR	B-Gene_or_gene_product
can	O
sensitise	O
paclitaxel	B-Simple_chemical
-	O
resistant	O
cervical	B-Cell
cancer	I-Cell
cells	I-Cell
.	O

Circulating	B-Cell
endothelial	I-Cell
cells	I-Cell
in	O
malignant	B-Cancer
disease	I-Cancer
.	O

Cancer	B-Cancer
is	O
a	O
disease	O
largely	O
dependent	O
on	O
neoangiogenesis	O
.	O

Cancer	B-Cancer
neoangiogenesis	O
is	O
often	O
disordered	O
and	O
abnormal	O
,	O
with	O
evidence	O
of	O
coexisting	O
vascular	B-Tissue
endothelial	I-Tissue
dysfunction	O
.	O

A	O
novel	O
method	O
of	O
assessing	O
vascular	B-Tissue
endothelial	I-Tissue
function	O
in	O
cancer	B-Cancer
is	O
via	O
the	O
quantification	O
of	O
circulating	B-Cell
endothelial	I-Cell
cells	I-Cell
(	O
CEC	B-Cell
)	O
.	O

Unusual	O
in	O
healthy	O
individuals	O
,	O
their	O
presence	O
in	O
elevated	O
numbers	O
often	O
indicates	O
substantial	O
vascular	B-Tissue
endothelial	I-Tissue
perturbation	O
.	O

Another	O
interesting	O
cell	B-Cell
type	O
is	O
the	O
endothelial	B-Cell
progenitor	I-Cell
cell	I-Cell
(	O
EPC	B-Cell
)	O
,	O
whose	O
numbers	O
increase	O
in	O
the	O
presence	O
of	O
vascular	B-Multi-tissue_structure
damage	O
.	O

Recent	O
research	O
suggests	O
that	O
EPCs	B-Cell
have	O
an	O
important	O
role	O
in	O
tumor	B-Cancer
vasculogenesis	O
.	O

Another	O
marker	O
being	O
investigated	O
in	O
the	O
context	O
of	O
vascular	B-Multi-tissue_structure
dysfunction	O
and	O
coagulopathy	O
is	O
the	O
endothelial	B-Cellular_component
microparticle	I-Cellular_component
(	O
EMP	B-Cellular_component
)	O
.	O

Thus	O
,	O
CECs	B-Cell
,	O
EPCs	B-Cell
and	O
EMPs	B-Cellular_component
may	O
represent	O
potentially	O
novel	O
methods	O
for	O
evaluating	O
the	O
vascular	B-Multi-tissue_structure
status	O
of	O
cancer	B-Cancer
patients	B-Organism
.	O

This	O
review	O
will	O
summarize	O
the	O
current	O
position	O
of	O
CECs	B-Cell
,	O
EPCs	B-Cell
and	O
EMPs	B-Cellular_component
in	O
cell	B-Cell
biology	O
terms	O
,	O
with	O
particular	O
emphasis	O
on	O
their	O
relationship	O
to	O
malignant	B-Cancer
disease	I-Cancer
.	O

GDP	B-Simple_chemical
and	O
AGE	B-Simple_chemical
receptors	O
:	O
mechanisms	O
of	O
peritoneal	B-Multi-tissue_structure
damage	O
.	O

Long	O
-	O
term	O
peritoneal	B-Multi-tissue_structure
dialysis	O
(	O
PD	O
)	O
is	O
limited	O
by	O
morphological	O
changes	O
of	O
the	O
peritoneal	B-Multi-tissue_structure
membrane	I-Multi-tissue_structure
.	O

Structural	O
changes	O
were	O
promoted	O
by	O
toxicity	O
of	O
glucose	B-Simple_chemical
degradation	I-Simple_chemical
products	I-Simple_chemical
(	O
GDPs	B-Simple_chemical
)	O
which	O
are	O
generated	O
during	O
heat	O
sterilization	O
in	O
peritoneal	B-Multi-tissue_structure
dialysis	O
fluids	O
(	O
PDFs	O
)	O
.	O

Besides	O
their	O
direct	O
toxicity	O
GDPs	B-Simple_chemical
promote	O
formation	O
of	O
advanced	B-Simple_chemical
glycation	I-Simple_chemical
endproducts	I-Simple_chemical
(	O
AGEs	B-Simple_chemical
)	O
.	O

RAGE	B-Gene_or_gene_product
(	O
receptor	B-Gene_or_gene_product
for	I-Gene_or_gene_product
AGE	I-Gene_or_gene_product
)	O
is	O
the	O
best	O
characterized	O
signal	O
transduction	O
receptor	O
for	O
AGEs	B-Simple_chemical
and	O
is	O
expressed	O
on	O
mesothelial	B-Cell
cells	I-Cell
.	O

The	O
effects	O
of	O
PDFs	O
with	O
different	O
amounts	O
of	O
GDPs	B-Simple_chemical
were	O
compared	O
on	O
morphological	O
changes	O
in	O
the	O
peritoneal	B-Multi-tissue_structure
membrane	I-Multi-tissue_structure
in	O
a	O
RAGE	B-Gene_or_gene_product
-	O
/	O
-	O
mouse	O
model	O
.	O

It	O
could	O
be	O
demonstrated	O
that	O
RAGE	B-Gene_or_gene_product
plays	O
a	O
pivotal	O
role	O
in	O
structural	O
damage	O
(	O
e	O
.	O
g	O
.	O
inflammation	O
,	O
neoangiogenesis	O
and	O
fibrosis	O
)	O
of	O
the	O
peritoneal	B-Multi-tissue_structure
membrane	I-Multi-tissue_structure
.	O

Further	O
investigations	O
of	O
this	O
pathway	O
with	O
regard	O
to	O
preventing	O
peritoneal	B-Multi-tissue_structure
fibrosis	O
should	O
be	O
performed	O
to	O
maintain	O
the	O
integrity	O
of	O
the	O
peritoneal	B-Multi-tissue_structure
membrane	I-Multi-tissue_structure
in	O
peritoneal	B-Multi-tissue_structure
dialysis	O
patients	B-Organism
.	O

Hyperfibrinogenemia	O
is	O
associated	O
with	O
lymphatic	B-Multi-tissue_structure
as	O
well	O
as	O
hematogenous	O
metastasis	O
and	O
worse	O
clinical	O
outcome	O
in	O
T2	B-Cancer
gastric	I-Cancer
cancer	I-Cancer
.	O

BACKGROUND	O
:	O
Abnormal	O
hemostasis	O
in	O
cancer	B-Cancer
patients	B-Organism
has	O
previously	O
been	O
described	O
,	O
however	O
the	O
correlation	O
between	O
the	O
plasma	B-Organism_substance
fibrinogen	B-Gene_or_gene_product
level	O
and	O
cancer	B-Cancer
metastasis	O
and	O
prognosis	O
has	O
not	O
been	O
reported	O
in	O
a	O
large	O
-	O
scale	O
clinical	O
study	O
.	O

METHODS	O
:	O
Preoperative	O
plasma	B-Organism_substance
fibrinogen	B-Gene_or_gene_product
levels	O
were	O
retrospectively	O
examined	O
in	O
405	O
patients	B-Organism
who	O
underwent	O
surgery	O
for	O
advanced	O
gastric	B-Cancer
cancer	I-Cancer
.	O

The	O
association	O
of	O
fibrinogen	B-Gene_or_gene_product
levels	O
with	O
clinical	O
/	O
pathological	O
findings	O
and	O
clinical	O
outcome	O
was	O
evaluated	O
.	O

RESULTS	O
:	O
There	O
was	O
a	O
positive	O
correlation	O
between	O
plasma	B-Organism_substance
fibrinogen	B-Gene_or_gene_product
levels	O
and	O
the	O
depth	O
of	O
invasion	O
(	O
p	O
<	O
0	O
.	O
05	O
)	O
.	O

Hyperfibrinogenemia	O
(	O
>	O
310	O
mg	O
/	O
dl	O
)	O
was	O
independently	O
associated	O
with	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
(	O
Odds	O
Ratio	O
;	O
2	O
.	O
342	O
,	O
P	O
=	O
0	O
.	O
0032	O
)	O
and	O
liver	B-Organ
(	O
Odds	O
Ratio	O
;	O
2	O
.	O
933	O
,	O
P	O
=	O
0	O
.	O
0147	O
)	O
metastasis	O
,	O
not	O
with	O
peritoneal	B-Multi-tissue_structure
metastasis	O
in	O
this	O
series	O
.	O

Patients	B-Organism
with	O
hyperfibrinogenemia	O
showed	O
worse	O
clinical	O
outcome	O
in	O
T2	B-Cancer
gastric	I-Cancer
cancer	I-Cancer
,	O
however	O
,	O
there	O
was	O
no	O
correlation	O
of	O
plasma	B-Organism_substance
fibrinogen	B-Gene_or_gene_product
level	O
with	O
prognosis	O
in	O
T3	B-Cancer
/	I-Cancer
T4	I-Cancer
gastric	I-Cancer
cancer	I-Cancer
.	O

CONCLUSION	O
:	O
Our	O
results	O
might	O
support	O
the	O
idea	O
that	O
hyperfibrinogenemia	O
can	O
augment	O
lymphatic	B-Cell
and	O
hematogeneous	O
metastasis	O
of	O
advanced	O
gastric	B-Cancer
cancer	I-Cancer
,	O
which	O
is	O
major	O
determinant	O
of	O
the	O
prognosis	O
in	O
T2	B-Cancer
gastric	I-Cancer
cancer	I-Cancer
.	O

Therefore	O
,	O
in	O
the	O
situation	O
without	O
peritoneal	B-Multi-tissue_structure
involvement	O
,	O
hyperfibrinogenemia	O
is	O
a	O
useful	O
biomarker	O
to	O
predict	O
the	O
possible	O
metastasis	O
and	O
worse	O
clinical	O
outcome	O
in	O
T2	B-Cancer
gastric	I-Cancer
cancer	I-Cancer
.	O

Defective	O
expression	O
of	O
HRK	B-Gene_or_gene_product
is	O
associated	O
with	O
promoter	O
methylation	O
in	O
primary	O
central	B-Cancer
nervous	I-Cancer
system	I-Cancer
lymphomas	I-Cancer
.	O

OBJECTIVES	O
:	O
Recently	O
,	O
it	O
has	O
been	O
reported	O
that	O
expression	O
of	O
the	O
HRK	B-Gene_or_gene_product
gene	O
was	O
significantly	O
reduced	O
by	O
hypermethylation	O
in	O
astrocytic	B-Cancer
tumors	I-Cancer
.	O

Our	O
aim	O
is	O
to	O
verify	O
the	O
alterations	O
in	O
the	O
HRK	B-Gene_or_gene_product
gene	O
in	O
primary	B-Cancer
central	I-Cancer
nervous	I-Cancer
system	I-Cancer
lymphomas	I-Cancer
(	O
PCNSLs	B-Cancer
)	O
.	O

METHODS	O
:	O
We	O
analyzed	O
the	O
hypermethylation	O
status	O
and	O
expression	O
of	O
the	O
gene	O
and	O
12q13	O
.	O
1	O
loss	O
of	O
heterozygosity	O
in	O
31	O
PCNSLs	B-Cancer
.	O

RESULTS	O
:	O
A	O
total	O
of	O
13	O
PCNSLs	B-Cancer
(	O
31	O
%	O
)	O
demonstrated	O
hypermethylation	O
in	O
either	O
the	O
promoter	O
or	O
exon	O
1	O
;	O
loss	O
of	O
HRK	B-Gene_or_gene_product
expression	O
was	O
immunohistochemically	O
observed	O
in	O
9	O
tumors	B-Cancer
and	O
was	O
significantly	O
associated	O
with	O
promoter	O
methylation	O
.	O

In	O
addition	O
,	O
higher	O
apoptotic	O
counts	O
were	O
associated	O
with	O
HRK	B-Gene_or_gene_product
positivity	O
.	O

PCNSLs	B-Cancer
with	O
HRK	B-Gene_or_gene_product
methylation	O
also	O
showed	O
methylation	O
of	O
multiple	O
genes	O
,	O
such	O
as	O
p14ARF	B-Gene_or_gene_product
,	O
p16INK4a	B-Gene_or_gene_product
,	O
RB1	B-Gene_or_gene_product
,	O
p27Kip1	B-Gene_or_gene_product
and	O
O6	B-Gene_or_gene_product
-	I-Gene_or_gene_product
MGMT	I-Gene_or_gene_product
.	O

Patients	B-Organism
with	O
tumors	B-Cancer
demonstrating	O
concurrent	O
methylation	O
of	O
more	O
than	O
half	O
of	O
their	O
genes	O
demonstrated	O
significantly	O
poorer	O
survival	O
and	O
earlier	O
recurrence	O
.	O

Hypermethylation	O
of	O
the	O
HRK	B-Gene_or_gene_product
promoter	O
alone	O
was	O
not	O
associated	O
with	O
overall	O
outcome	O
,	O
but	O
relapse	O
-	O
free	O
survival	O
was	O
significantly	O
shorter	O
.	O

CONCLUSIONS	O
:	O
Our	O
findings	O
suggest	O
that	O
transcriptional	O
repression	O
of	O
HRK	B-Gene_or_gene_product
is	O
caused	O
by	O
promoter	O
hypermethylation	O
in	O
PCNSL	B-Cancer
,	O
and	O
that	O
the	O
loss	O
of	O
HRK	B-Gene_or_gene_product
associated	O
with	O
the	O
methylation	O
profile	O
of	O
other	O
genes	O
is	O
a	O
potential	O
step	O
in	O
the	O
modulation	O
of	O
cellular	B-Cell
death	O
by	O
apoptosis	O
during	O
PCNSL	B-Cancer
tumorigenesis	O
.	O

Differential	O
mechanisms	O
of	O
radiosensitization	O
by	O
2	B-Simple_chemical
-	I-Simple_chemical
deoxy	I-Simple_chemical
-	I-Simple_chemical
D	I-Simple_chemical
-	I-Simple_chemical
glucose	I-Simple_chemical
in	O
the	O
monolayers	B-Cell
and	O
multicellular	B-Cell
spheroids	I-Cell
of	O
a	O
human	B-Organism
glioma	B-Cell
cell	I-Cell
line	I-Cell
.	O

In	O
vitro	O
studies	O
using	O
monolayer	B-Cell
cultures	I-Cell
of	O
human	B-Organism
tumor	B-Cell
cell	I-Cell
lines	I-Cell
have	O
shown	O
that	O
2	B-Simple_chemical
-	I-Simple_chemical
DG	I-Simple_chemical
selectively	O
inhibits	O
energy	O
-	O
dependent	O
DNA	B-Cellular_component
repair	O
and	O
cellular	B-Cell
recovery	O
processes	O
in	O
cancer	B-Cell
cells	I-Cell
.	O

However	O
,	O
monolayer	B-Cell
cultures	I-Cell
differ	O
greatly	O
from	O
the	O
complex	O
environmental	O
conditions	O
generated	O
in	O
solid	B-Cancer
tumors	I-Cancer
that	O
develop	O
inhomogeneous	B-Pathological_formation
hypoxic	I-Pathological_formation
and	O
necrotic	B-Pathological_formation
regions	I-Pathological_formation
.	O

In	O
contrast	O
,	O
multicellular	B-Cell
spheroids	I-Cell
mimic	O
heterogeneous	O
cellular	B-Cell
behavior	O
and	O
the	O
consequent	O
functional	O
characteristics	O
of	O
in	O
vivo	O
solid	B-Cancer
tumors	I-Cancer
,	O
and	O
serve	O
as	O
important	O
in	O
vitro	O
model	O
to	O
investigate	O
tumor	B-Cancer
biology	O
and	O
responses	O
to	O
potential	O
therapeutic	O
agents	O
.	O

The	O
present	O
study	O
compares	O
the	O
radiomodification	O
by	O
2	B-Simple_chemical
-	I-Simple_chemical
DG	I-Simple_chemical
in	O
monolayer	B-Cell
cultures	I-Cell
and	O
spheroids	B-Cell
of	O
a	O
human	B-Organism
glioma	B-Cell
cell	I-Cell
line	I-Cell
(	O
BMG	B-Cell
-	I-Cell
1	I-Cell
)	O
to	O
gain	O
insight	O
into	O
the	O
effects	O
in	O
solid	B-Cancer
tumors	I-Cancer
.	O

In	O
spheroids	B-Cell
,	O
the	O
glucose	B-Simple_chemical
consumption	O
(	O
2	O
.	O
1	O
p	O
mole	O
/	O
cell	O
/	O
h	O
)	O
and	O
lactate	B-Simple_chemical
production	O
(	O
3	O
.	O
67	O
p	O
mole	O
/	O
cell	O
/	O
h	O
)	O
was	O
nearly	O
2	O
-	O
3	O
fold	O
higher	O
than	O
in	O
monolayer	B-Cell
cells	I-Cell
(	O
0	O
.	O
83	O
and	O
1	O
.	O
43	O
p	O
mole	O
/	O
cell	O
/	O
h	O
respectively	O
)	O
.	O

Presence	O
of	O
2	B-Simple_chemical
-	I-Simple_chemical
DG	I-Simple_chemical
(	O
5	O
mM	O
)	O
for	O
2	O
-	O
4	O
h	O
inhibited	O
the	O
glucose	B-Simple_chemical
usage	O
and	O
lactate	B-Simple_chemical
production	O
by	O
70	O
%	O
in	O
spheroids	B-Cell
,	O
while	O
a	O
35	O
%	O
reduction	O
was	O
observed	O
in	O
monolayer	B-Cell
cells	I-Cell
.	O

Under	O
these	O
conditions	O
,	O
2	B-Simple_chemical
-	I-Simple_chemical
DG	I-Simple_chemical
drastically	O
enhanced	O
the	O
radiation	O
-	O
induced	O
cell	B-Cell
death	O
of	O
spheroids	B-Cell
(	O
by	O
2	O
-	O
3	O
folds	O
)	O
;	O
while	O
a	O
40	O
%	O
increase	O
was	O
observed	O
in	O
monolayer	B-Cell
cells	I-Cell
.	O

Radiosensitization	O
by	O
2	B-Simple_chemical
-	I-Simple_chemical
DG	I-Simple_chemical
in	O
monolayer	B-Cell
cells	I-Cell
was	O
primarily	O
due	O
to	O
an	O
increase	O
in	O
mitotic	O
death	O
(	O
23	O
%	O
)	O
linked	O
to	O
cytogenetic	O
damage	O
(	O
micronuclei	B-Cellular_component
)	O
,	O
whereas	O
a	O
profound	O
induction	O
of	O
apoptosis	O
(	O
40	O
%	O
)	O
accounted	O
for	O
the	O
sensitization	O
in	O
spheroids	B-Cell
.	O

Although	O
the	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
Bax	B-Gene_or_gene_product
levels	O
were	O
significantly	O
higher	O
in	O
spheroids	B-Cell
,	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
/	O
Bax	B-Gene_or_gene_product
ratio	O
was	O
similar	O
in	O
monolayers	B-Cell
and	O
spheroids	B-Cell
.	O

Comet	O
assay	O
revealed	O
a	O
late	O
onset	O
of	O
DNA	B-Cellular_component
breaks	O
in	O
the	O
presence	O
of	O
2	B-Simple_chemical
-	I-Simple_chemical
DG	I-Simple_chemical
following	O
irradiation	O
only	O
in	O
spheroids	B-Cell
,	O
which	O
corroborated	O
well	O
with	O
the	O
late	O
onset	O
of	O
oxidative	O
stress	O
.	O

2	B-Simple_chemical
-	I-Simple_chemical
DG	I-Simple_chemical
did	O
not	O
induce	O
a	O
significant	O
cell	B-Cell
cycle	O
delay	O
in	O
monolayers	B-Cell
,	O
while	O
a	O
transient	O
G	O
(	O
2	O
)	O
delay	O
was	O
apparent	O
in	O
spheroids	B-Cell
.	O

[	O
Autologous	O
bone	B-Cell
marrow	I-Cell
stem	I-Cell
cell	I-Cell
or	O
peripheral	B-Cell
blood	I-Cell
endothelial	I-Cell
progenitor	I-Cell
cell	I-Cell
therapy	O
in	O
patients	B-Organism
with	O
peripheral	O
limb	B-Organism_subdivision
ischaemia	O
]	O

No	O
effective	O
medical	O
therapies	O
have	O
been	O
developed	O
sofar	O
to	O
enhance	O
blood	B-Organism_substance
flow	O
in	O
the	O
legs	B-Organism_subdivision
of	O
patients	B-Organism
with	O
peripheral	B-Multi-tissue_structure
arterial	I-Multi-tissue_structure
disease	O
(	O
PAD	O
)	O
.	O

For	O
patients	B-Organism
with	O
limb	B-Organism_subdivision
threatening	O
ischaemia	O
the	O
only	O
option	O
for	O
relief	O
of	O
rest	O
pain	O
or	O
gangraena	O
is	O
amputation	O
.	O

There	O
is	O
evidence	O
in	O
experimental	O
and	O
clinical	O
studies	O
that	O
adult	O
bone	B-Cell
marrow	I-Cell
-	I-Cell
derived	I-Cell
stem	I-Cell
cells	I-Cell
and	O
endothelial	B-Cell
progenitor	I-Cell
cells	I-Cell
participate	O
in	O
the	O
development	O
of	O
new	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
,	O
called	O
neoangiogenesis	O
or	O
neovascularization	O
.	O

Clinical	O
results	O
induced	O
by	O
autologous	O
bone	B-Cell
marrow	I-Cell
stem	I-Cell
cells	I-Cell
or	O
angiogenic	O
growth	O
/	O
differentiation	O
factors	O
in	O
end	O
-	O
stage	O
patients	B-Organism
with	O
PAD	O
are	O
summarized	O
.	O

Considering	O
the	O
relatively	O
few	O
number	O
of	O
patients	B-Organism
treated	O
by	O
angiogenic	O
therapy	O
,	O
the	O
interpretation	O
of	O
clinical	O
results	O
needs	O
cautiousness	O
.	O

Formation	O
of	O
new	O
bone	B-Tissue
during	O
vertical	O
distraction	O
osteogenesis	O
of	O
the	O
human	B-Organism
mandible	B-Organ
is	O
related	O
to	O
the	O
presence	O
of	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
.	O

We	O
examined	O
the	O
effect	O
of	O
distraction	O
rate	O
on	O
blood	B-Multi-tissue_structure
vessel	I-Multi-tissue_structure
growth	O
in	O
intramembraneous	O
ossification	O
after	O
vertical	O
distraction	O
osteogenesis	O
in	O
the	O
human	B-Organism
mandible	B-Organ
.	O

Six	O
edentulous	O
patients	B-Organism
(	O
aged	O
60	O
+	O
/	O
-	O
9	O
years	O
)	O
with	O
a	O
severely	O
atrophic	O
mandible	B-Organ
underwent	O
bone	B-Tissue
augmentation	O
with	O
distraction	O
osteogenesis	O
.	O

Two	O
distraction	O
rates	O
(	O
0	O
.	O
5	O
and	O
1	O
mm	O
/	O
day	O
)	O
were	O
compared	O
and	O
for	O
each	O
group	O
three	O
patients	B-Organism
were	O
analyzed	O
.	O

Vascular	B-Multi-tissue_structure
histomorphometry	O
was	O
carried	O
out	O
in	O
two	O
different	O
areas	O
in	O
the	O
distraction	O
gap	O
:	O
(	O
1	O
)	O
in	O
the	O
first	O
and	O
(	O
2	O
)	O
in	O
the	O
second	O
1	O
mm	O
area	O
from	O
the	O
osteotomy	O
line	O
,	O
representing	O
the	O
oldest	O
and	O
younger	O
new	O
-	O
bone	B-Tissue
area	O
,	O
respectively	O
.	O

Correlation	O
analysis	O
was	O
performed	O
between	O
blood	B-Multi-tissue_structure
vessel	I-Multi-tissue_structure
parameters	O
and	O
the	O
amount	O
of	O
new	O
bone	B-Tissue
formed	O
during	O
distraction	O
.	O

Histological	O
analysis	O
demonstrated	O
the	O
presence	O
of	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
throughout	O
the	O
soft	B-Tissue
connective	I-Tissue
tissue	I-Tissue
in	O
the	O
distraction	O
gap	O
.	O

The	O
volume	O
density	O
of	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
between	O
the	O
two	O
investigated	O
areas	O
was	O
significantly	O
lower	O
in	O
the	O
1	O
mm	O
/	O
day	O
groups	O
,	O
suggesting	O
a	O
delay	O
in	O
angiogenesis	O
in	O
this	O
group	O
of	O
patients	B-Organism
.	O

A	O
positive	O
correlation	O
between	O
blood	B-Multi-tissue_structure
vessel	I-Multi-tissue_structure
volume	O
and	O
bone	B-Tissue
volume	O
density	O
was	O
found	O
in	O
the	O
younger	O
new	O
-	O
bone	B-Tissue
area	O
but	O
not	O
in	O
the	O
oldest	O
new	O
-	O
bone	B-Tissue
area	O
.	O

This	O
correlation	O
was	O
due	O
to	O
a	O
higher	O
number	O
of	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
rather	O
than	O
to	O
a	O
larger	O
size	O
of	O
the	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
.	O

Our	O
data	O
suggest	O
that	O
the	O
lower	O
blood	B-Multi-tissue_structure
vessel	I-Multi-tissue_structure
density	O
found	O
in	O
the	O
patients	B-Organism
with	O
1	O
mm	O
/	O
day	O
distraction	O
rate	O
may	O
be	O
related	O
to	O
disruption	O
of	O
angiogenesis	O
in	O
the	O
soft	B-Tissue
connective	I-Tissue
tissue	I-Tissue
of	O
the	O
gap	O
or	O
to	O
a	O
less	O
optimal	O
mechanical	O
stimulation	O
of	O
cells	B-Cell
involved	O
in	O
angiogenesis	O
.	O

This	O
probably	O
results	O
in	O
the	O
slower	O
rate	O
of	O
osteogenesis	O
seen	O
at	O
the	O
1	O
mm	O
/	O
day	O
distraction	O
rate	O
compared	O
with	O
the	O
0	O
.	O
5	O
mm	O
/	O
day	O
distraction	O
rate	O
.	O

The	O
data	O
support	O
the	O
concept	O
that	O
a	O
positive	O
relationship	O
exists	O
between	O
the	O
density	O
of	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
and	O
the	O
formation	O
of	O
bone	B-Tissue
.	O

For	O
distraction	O
of	O
the	O
human	B-Organism
mandible	B-Organ
in	O
elderly	O
patients	B-Organism
,	O
a	O
distraction	O
rate	O
of	O
0	O
.	O
5	O
mm	O
/	O
day	O
seems	O
beneficial	O
.	O

Neoplastic	O
transformation	O
of	O
ciliary	B-Tissue
body	I-Tissue
epithelium	I-Tissue
is	O
associated	O
with	O
loss	O
of	O
opticin	B-Gene_or_gene_product
expression	O
.	O

BACKGROUND	O
:	O
Opticin	B-Gene_or_gene_product
is	O
a	O
recently	O
discovered	O
glycoprotein	O
present	O
predominantly	O
in	O
the	O
vitreous	B-Organism_substance
humour	I-Organism_substance
.	O

It	O
is	O
synthesised	O
and	O
secreted	O
by	O
the	O
ciliary	B-Tissue
body	I-Tissue
epithelium	I-Tissue
(	O
CBE	B-Tissue
)	O
from	O
the	O
initiation	O
of	O
CBE	B-Tissue
development	O
in	O
the	O
embryo	B-Developing_anatomical_structure
,	O
and	O
production	O
continues	O
throughout	O
life	O
.	O

AIM	O
:	O
To	O
determine	O
whether	O
a	O
variety	O
of	O
ciliary	B-Cancer
body	I-Cancer
tumours	I-Cancer
synthesise	O
opticin	B-Gene_or_gene_product
to	O
characterise	O
further	O
its	O
role	O
in	O
ciliary	B-Multi-tissue_structure
body	I-Multi-tissue_structure
health	O
and	O
disease	O
.	O

METHODS	O
:	O
Immunohistochemistry	O
was	O
used	O
to	O
determine	O
the	O
distribution	O
of	O
opticin	B-Gene_or_gene_product
in	O
normal	O
human	B-Organism
CBE	B-Tissue
,	O
and	O
in	O
hyperplastic	B-Cancer
and	O
neoplastic	B-Cancer
CBE	I-Cancer
lesions	I-Cancer
.	O

RESULTS	O
:	O
Opticin	B-Gene_or_gene_product
was	O
immunolocalised	O
to	O
the	O
basal	B-Cellular_component
cell	I-Cellular_component
surface	I-Cellular_component
and	O
basement	B-Cellular_component
membrane	I-Cellular_component
material	I-Cellular_component
of	O
the	O
non	B-Tissue
-	I-Tissue
pigmented	I-Tissue
CBE	I-Tissue
in	O
nine	O
donor	O
eyes	B-Organ
as	O
well	O
as	O
four	O
hyperplastic	B-Cancer
lesions	I-Cancer
of	O
the	O
CBE	B-Tissue
(	O
Fuchs	B-Cancer
'	I-Cancer
s	I-Cancer
adenoma	I-Cancer
)	O
.	O

By	O
contrast	O
,	O
none	O
of	O
eight	O
neoplastic	B-Cancer
lesions	I-Cancer
(	O
two	O
adenoma	B-Cancer
and	O
six	O
adenocarcinoma	B-Cancer
)	O
of	O
CBE	B-Tissue
stained	O
for	O
opticin	B-Gene_or_gene_product
.	O

CONCLUSION	O
:	O
The	O
present	O
series	O
supports	O
the	O
theory	O
that	O
opticin	B-Gene_or_gene_product
is	O
produced	O
by	O
the	O
non	B-Tissue
-	I-Tissue
pigmented	I-Tissue
CBE	I-Tissue
throughout	O
adult	O
life	O
.	O

Loss	O
of	O
opticin	B-Gene_or_gene_product
expression	O
by	O
this	O
tissue	B-Tissue
is	O
associated	O
with	O
and	O
could	O
contribute	O
towards	O
neoplastic	O
transformation	O
.	O

Collagen	B-Gene_or_gene_product
-	O
poly	B-Simple_chemical
glycolic	I-Simple_chemical
acid	I-Simple_chemical
hybrid	O
matrix	O
with	O
basic	B-Gene_or_gene_product
fibroblast	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
accelerated	O
angiogenesis	O
and	O
granulation	B-Tissue
tissue	I-Tissue
formation	O
in	O
diabetic	O
mice	B-Organism
.	O

Because	O
poor	O
skin	B-Pathological_formation
wound	I-Pathological_formation
healing	O
associated	O
with	O
diabetes	O
is	O
thought	O
to	O
be	O
partly	O
a	O
result	O
from	O
impaired	O
angiogenesis	O
,	O
treatments	O
that	O
improve	O
angiogenesis	O
could	O
have	O
important	O
clinical	O
applications	O
.	O

We	O
herein	O
report	O
the	O
effects	O
of	O
novel	O
developed	O
material	O
,	O
collagen	B-Gene_or_gene_product
-	O
poly	B-Simple_chemical
glycolic	I-Simple_chemical
acid	I-Simple_chemical
fiber	O
hybrid	O
matrix	O
,	O
being	O
used	O
together	O
with	O
basic	B-Gene_or_gene_product
fibroblast	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
to	O
promote	O
wound	B-Pathological_formation
healing	O
of	O
full	O
-	O
thickness	O
skin	B-Organ
defects	O
on	O
the	O
back	O
of	O
type	O
2	O
diabetic	O
Lepr	B-Gene_or_gene_product
(	O
db	O
)	O
mice	O
.	O

Our	O
data	O
indicates	O
that	O
this	O
therapeutic	O
approach	O
markedly	O
promotes	O
angiogenesis	O
and	O
granulation	B-Tissue
tissue	I-Tissue
formation	O
in	O
comparison	O
with	O
other	O
conditions	O
14	O
days	O
after	O
wounding	O
.	O

p18Ink4c	B-Gene_or_gene_product
,	O
but	O
not	O
p27Kip1	B-Gene_or_gene_product
,	O
collaborates	O
with	O
Men1	B-Gene_or_gene_product
to	O
suppress	O
neuroendocrine	B-Cancer
organ	I-Cancer
tumors	I-Cancer
.	O

Mutant	O
mice	B-Organism
lacking	O
both	O
cyclin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
dependent	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
(	O
CDK	B-Gene_or_gene_product
)	O
inhibitors	O
p18	B-Gene_or_gene_product
(	O
Ink4c	B-Gene_or_gene_product
)	O
and	O
p27	B-Gene_or_gene_product
(	O
Kip1	B-Gene_or_gene_product
)	O
develop	O
a	O
tumor	B-Cancer
spectrum	O
reminiscent	O
of	O
human	B-Organism
multiple	B-Cancer
endocrine	I-Cancer
neoplasia	I-Cancer
(	O
MEN	B-Cancer
)	O
syndromes	O
.	O

To	O
determine	O
how	O
p18	B-Gene_or_gene_product
and	O
p27	B-Gene_or_gene_product
genetically	O
interact	O
with	O
Men1	B-Gene_or_gene_product
,	O
the	O
tumor	B-Cancer
suppressor	O
gene	O
mutated	O
in	O
familial	O
MEN1	B-Cancer
,	O
we	O
characterized	O
p18	B-Gene_or_gene_product
-	O
Men1	B-Gene_or_gene_product
and	O
p27	B-Gene_or_gene_product
-	O
Men1	B-Gene_or_gene_product
double	O
mutant	O
mice	O
.	O

Compared	O
with	O
their	O
corresponding	O
single	B-Organism
mutant	I-Organism
littermates	I-Organism
,	O
the	O
p18	B-Gene_or_gene_product
(	O
-	O
/	O
-	O
)	O
;	O
Men1	B-Gene_or_gene_product
(	O
+	O
/	O
-	O
)	O
mice	O
develop	O
tumors	B-Cancer
at	O
an	O
accelerated	O
rate	O
and	O
with	O
an	O
increased	O
incidence	O
in	O
the	O
pituitary	B-Organ
,	O
thyroid	B-Organ
,	O
parathyroid	B-Organ
,	O
and	O
pancreas	B-Organ
.	O

In	O
the	O
pituitary	B-Multi-tissue_structure
and	O
pancreatic	B-Multi-tissue_structure
islets	I-Multi-tissue_structure
,	O
phosphorylation	O
of	O
the	O
retinoblastoma	B-Gene_or_gene_product
(	O
Rb	B-Gene_or_gene_product
)	O
protein	O
at	O
both	O
CDK2	B-Gene_or_gene_product
and	O
CDK4	B-Gene_or_gene_product
/	I-Gene_or_gene_product
6	I-Gene_or_gene_product
sites	O
was	O
increased	O
in	O
p18	B-Gene_or_gene_product
(	I-Gene_or_gene_product
-	I-Gene_or_gene_product
/	I-Gene_or_gene_product
-	I-Gene_or_gene_product
)	I-Gene_or_gene_product
and	O
Men1	B-Gene_or_gene_product
(	O
+	O
/	O
-	O
)	O
cells	O
and	O
was	O
further	O
increased	O
in	O
p18	B-Gene_or_gene_product
(	O
-	O
/	O
-	O
)	O
;	O
Men1	B-Gene_or_gene_product
(	O
+	O
/	O
-	O
)	O
cells	O
.	O

The	O
remaining	O
wild	O
-	O
type	O
Men1	B-Gene_or_gene_product
allele	O
was	O
lost	O
in	O
most	O
tumors	B-Cancer
from	O
Men1	B-Gene_or_gene_product
(	O
+	O
/	O
-	O
)	O
mice	O
but	O
was	O
retained	O
in	O
most	O
tumors	B-Cancer
from	O
p18	B-Gene_or_gene_product
(	O
-	O
/	O
-	O
)	O
;	O
Men1	B-Gene_or_gene_product
(	O
+	O
/	O
-	O
)	O
mice	O
.	O

Combined	O
mutations	O
of	O
p27	B-Gene_or_gene_product
(	O
-	O
/	O
-	O
)	O
and	O
Men1	B-Gene_or_gene_product
(	O
+	O
/	O
-	O
)	O
,	O
in	O
contrast	O
,	O
did	O
not	O
exhibit	O
noticeable	O
synergistic	O
stimulation	O
of	O
Rb	B-Gene_or_gene_product
kinase	O
activity	O
,	O
cell	B-Cell
proliferation	O
,	O
and	O
tumor	B-Cancer
growth	O
.	O

These	O
results	O
demonstrate	O
that	O
functional	O
collaboration	O
exists	O
between	O
p18	B-Gene_or_gene_product
and	O
Men1	B-Gene_or_gene_product
and	O
suggest	O
that	O
Men1	B-Gene_or_gene_product
may	O
regulate	O
additional	O
factor	O
(	O
s	O
)	O
that	O
interact	O
with	O
p18	B-Gene_or_gene_product
and	O
p27	B-Gene_or_gene_product
differently	O
.	O

Punica	B-Organism
granatum	I-Organism
(	O
pomegranate	B-Organism
)	O
and	O
its	O
potential	O
for	O
prevention	O
and	O
treatment	O
of	O
inflammation	O
and	O
cancer	B-Cancer
.	O

The	O
last	O
7	O
years	O
have	O
seen	O
over	O
seven	O
times	O
as	O
many	O
publications	O
indexed	O
by	O
Medline	O
dealing	O
with	O
pomegranate	B-Organism
and	O
Punica	B-Organism
granatum	I-Organism
than	O
in	O
all	O
the	O
years	O
preceding	O
them	O
.	O

Because	O
of	O
this	O
,	O
and	O
the	O
virtual	O
explosion	O
of	O
interest	O
in	O
pomegranate	B-Organism
as	O
a	O
medicinal	O
and	O
nutritional	O
product	O
that	O
has	O
followed	O
,	O
this	O
review	O
is	O
accordingly	O
launched	O
.	O

The	O
pomegranate	B-Organism
tree	B-Organism_subdivision
,	O
Punica	B-Organism
granatum	I-Organism
,	O
especially	O
its	O
fruit	O
,	O
possesses	O
a	O
vast	O
ethnomedical	O
history	O
and	O
represents	O
a	O
phytochemical	O
reservoir	O
of	O
heuristic	O
medicinal	O
value	O
.	O

The	O
tree	O
/	O
fruit	O
can	O
be	O
divided	O
into	O
several	O
anatomical	O
compartments	O
:	O
(	O
1	O
)	O
seed	B-Developing_anatomical_structure
,	O
(	O
2	O
)	O
juice	B-Organism_substance
,	O
(	O
3	O
)	O
peel	B-Tissue
,	O
(	O
4	O
)	O
leaf	B-Organ
,	O
(	O
5	O
)	O
flower	B-Organism_subdivision
,	O
(	O
6	O
)	O
bark	B-Tissue
,	O
and	O
(	O
7	O
)	O
roots	B-Organ
,	O
each	O
of	O
which	O
has	O
interesting	O
pharmacologic	O
activity	O
.	O

Juice	B-Organism_substance
and	O
peels	B-Tissue
,	O
for	O
example	O
,	O
possess	O
potent	O
antioxidant	O
properties	O
,	O
while	O
juice	B-Organism_substance
,	O
peel	B-Tissue
and	O
oil	B-Organism_substance
are	O
all	O
weakly	O
estrogenic	O
and	O
heuristically	O
of	O
interest	O
for	O
the	O
treatment	O
of	O
menopausal	O
symptoms	O
and	O
sequellae	O
.	O

The	O
use	O
of	O
juice	B-Organism_substance
,	O
peel	B-Tissue
and	O
oil	B-Organism_substance
have	O
also	O
been	O
shown	O
to	O
possess	O
anticancer	B-Cancer
activities	O
,	O
including	O
interference	O
with	O
tumor	B-Cell
cell	I-Cell
proliferation	O
,	O
cell	B-Cell
cycle	O
,	O
invasion	O
and	O
angiogenesis	O
.	O

These	O
may	O
be	O
associated	O
with	O
plant	O
based	O
anti	O
-	O
inflammatory	O
effects	O
,	O
The	O
phytochemistry	O
and	O
pharmacological	O
actions	O
of	O
all	O
Punica	B-Organism
granatum	I-Organism
components	O
suggest	O
a	O
wide	O
range	O
of	O
clinical	O
applications	O
for	O
the	O
treatment	O
and	O
prevention	O
of	O
cancer	B-Cancer
,	O
as	O
well	O
as	O
other	O
diseases	O
where	O
chronic	O
inflammation	O
is	O
believed	O
to	O
play	O
an	O
essential	O
etiologic	O
role	O
.	O

Recombinant	O
human	B-Organism
prothrombin	B-Gene_or_gene_product
kringle	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
inhibits	O
B16F10	B-Cancer
melanoma	I-Cancer
metastasis	O
through	O
inhibition	O
of	O
neovascularization	O
and	O
reduction	O
of	O
matrix	B-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
expression	O
.	O

Angiogenesis	O
,	O
a	O
multi	O
-	O
step	O
process	O
which	O
involves	O
endothelial	B-Cell
cell	I-Cell
proliferation	O
,	O
adhesion	O
,	O
migration	O
,	O
and	O
basement	B-Cellular_component
membrane	I-Cellular_component
(	O
BM	B-Cellular_component
)	O
degradation	O
,	O
is	O
essential	O
for	O
tumor	B-Cancer
metastasis	O
.	O

Here	O
we	O
show	O
that	O
recombinant	O
human	B-Organism
prothrombin	B-Gene_or_gene_product
kringle	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
(	O
rk	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
)	O
inhibited	O
bovine	B-Organism
capillary	B-Cell
endothelial	I-Cell
cell	I-Cell
migration	O
with	O
an	O
IC	O
(	O
50	O
)	O
(	O
concentration	O
for	O
half	O
maximal	O
inhibition	O
)	O
of	O
38	O
nM	O
and	O
inhibited	O
adhesion	O
to	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
(	O
ECM	B-Cellular_component
)	O
proteins	O
.	O

Because	O
tumor	B-Cancer
metastasis	O
requires	O
angiogenesis	O
,	O
we	O
examined	O
whether	O
rk	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
could	O
inhibit	O
metastases	O
induced	O
by	O
injection	O
of	O
B16F10	B-Cell
melanoma	I-Cell
cells	I-Cell
into	O
mice	B-Organism
.	O

The	O
results	O
revealed	O
that	O
the	O
metastatic	O
tumors	B-Cancer
in	O
mouse	B-Organism
lung	B-Organ
were	O
markedly	O
decreased	O
in	O
a	O
dose	O
-	O
dependent	O
manner	O
and	O
acute	O
lung	B-Organ
injury	O
induced	O
by	O
B16F10	B-Cancer
melanoma	I-Cancer
metastasis	O
was	O
diminished	O
by	O
systemic	O
rk	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
treatment	O
.	O

In	O
immunohistochemical	O
analysis	O
,	O
rk	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
reduced	O
expression	O
of	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
,	O
which	O
is	O
a	O
potent	O
angiogenic	O
activator	O
and	O
neovascularization	O
in	O
the	O
mouse	B-Organism
lung	B-Organ
.	O

Also	O
,	O
rk	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
diminished	O
the	O
expression	O
of	O
matrix	B-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
-	B-Gene_or_gene_product
9	I-Gene_or_gene_product
in	O
the	O
mouse	B-Organism
lung	B-Organ
which	O
induces	O
tumor	B-Cancer
metastasis	O
and	O
angiogenesis	O
.	O

These	O
data	O
suggest	O
that	O
inhibition	O
of	O
B16F10	B-Cancer
melanoma	I-Cancer
metastasis	O
by	O
rk	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
was	O
caused	O
by	O
inhibition	O
of	O
neovascularization	O
and	O
reduction	O
of	O
matrix	B-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
expression	O
.	O

Activation	O
of	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
through	O
reactive	B-Simple_chemical
oxygen	I-Simple_chemical
species	I-Simple_chemical
mediates	O
20	B-Simple_chemical
-	I-Simple_chemical
hydroxyeicosatetraenoic	I-Simple_chemical
acid	I-Simple_chemical
-	O
induced	O
endothelial	B-Cell
cell	I-Cell
proliferation	O
.	O

20	B-Simple_chemical
-	I-Simple_chemical
Hydroxyeicosatetraenoic	I-Simple_chemical
acid	I-Simple_chemical
(	O
20	B-Simple_chemical
-	I-Simple_chemical
HETE	I-Simple_chemical
)	O
is	O
formed	O
by	O
the	O
omega	O
-	O
hydroxylation	O
of	O
arachidonic	B-Simple_chemical
acid	I-Simple_chemical
by	O
cytochrome	B-Gene_or_gene_product
P450	I-Gene_or_gene_product
4A	I-Gene_or_gene_product
and	O
4F	B-Gene_or_gene_product
enzymes	O
,	O
and	O
it	O
induces	O
angiogenic	O
responses	O
in	O
vivo	O
.	O

To	O
test	O
the	O
hypothesis	O
that	O
20	B-Simple_chemical
-	I-Simple_chemical
HETE	I-Simple_chemical
increases	O
endothelial	B-Cell
cell	I-Cell
(	O
EC	B-Cell
)	O
proliferation	O
via	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
,	O
we	O
studied	O
the	O
effects	O
of	O
WIT003	B-Simple_chemical
[	O
20	B-Simple_chemical
-	I-Simple_chemical
hydroxyeicosa	I-Simple_chemical
-	I-Simple_chemical
5	I-Simple_chemical
(	I-Simple_chemical
Z	I-Simple_chemical
)	I-Simple_chemical
,	I-Simple_chemical
14	I-Simple_chemical
(	I-Simple_chemical
Z	I-Simple_chemical
)	I-Simple_chemical
-	I-Simple_chemical
dienoic	I-Simple_chemical
acid	I-Simple_chemical
]	O
,	O
a	O
20	B-Simple_chemical
-	I-Simple_chemical
HETE	I-Simple_chemical
analog	O
on	O
human	B-Organism
macrovascular	B-Cell
or	O
microvascular	B-Cell
EC	I-Cell
.	O

WIT003	B-Simple_chemical
,	O
as	O
well	O
as	O
pure	O
20	B-Simple_chemical
-	I-Simple_chemical
HETE	I-Simple_chemical
,	O
stimulated	O
EC	B-Cell
proliferation	O
by	O
approximately	O
40	O
%	O
.	O

These	O
proliferative	O
effects	O
were	O
accompanied	O
by	O
increased	O
VEGF	B-Gene_or_gene_product
expression	O
and	O
release	O
that	O
were	O
observed	O
as	O
early	O
as	O
4	O
h	O
after	O
20	B-Simple_chemical
-	I-Simple_chemical
HETE	I-Simple_chemical
agonist	O
addition	O
.	O

This	O
was	O
accompanied	O
by	O
increased	O
phosphorylation	O
of	O
the	O
VEGF	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
2	I-Gene_or_gene_product
.	O

The	O
proliferative	O
effects	O
of	O
20	B-Simple_chemical
-	I-Simple_chemical
HETE	I-Simple_chemical
were	O
markedly	O
inhibited	O
by	O
a	O
VEGF	B-Gene_or_gene_product
-	O
neutralizing	O
antibody	O
.	O

Polyethylene	B-Simple_chemical
glycol	I-Simple_chemical
-	I-Simple_chemical
superoxide	I-Simple_chemical
dismutase	I-Simple_chemical
(	O
PEG	B-Simple_chemical
-	I-Simple_chemical
SOD	I-Simple_chemical
)	O
markedly	O
inhibited	O
both	O
the	O
increases	O
in	O
VEGF	B-Gene_or_gene_product
expression	O
and	O
the	O
proliferative	O
effects	O
of	O
20	B-Simple_chemical
-	I-Simple_chemical
HETE	I-Simple_chemical
.	O

In	O
contrast	O
,	O
administration	O
of	O
the	O
NAD	B-Gene_or_gene_product
(	I-Gene_or_gene_product
P	I-Gene_or_gene_product
)	I-Gene_or_gene_product
H	I-Gene_or_gene_product
oxidase	I-Gene_or_gene_product
inhibitor	O
apocynin	B-Simple_chemical
had	O
no	O
effect	O
to	O
the	O
proliferative	O
response	O
to	O
20	B-Simple_chemical
-	I-Simple_chemical
HETE	I-Simple_chemical
.	O

The	O
20	B-Simple_chemical
-	I-Simple_chemical
HETE	I-Simple_chemical
agonist	O
markedly	O
increased	O
superoxide	B-Simple_chemical
formation	O
as	O
reflected	O
by	O
an	O
increase	O
in	O
dihydroethidium	B-Simple_chemical
staining	O
of	O
EC	B-Cell
,	O
and	O
this	O
increase	O
was	O
inhibited	O
by	O
PEG	B-Simple_chemical
-	I-Simple_chemical
SOD	I-Simple_chemical
but	O
not	O
by	O
apocynin	B-Simple_chemical
.	O

20	B-Simple_chemical
-	I-Simple_chemical
HETE	I-Simple_chemical
also	O
increased	O
the	O
phosphorylation	O
of	O
p42	B-Gene_or_gene_product
/	I-Gene_or_gene_product
p44	I-Gene_or_gene_product
mitogen	I-Gene_or_gene_product
-	I-Gene_or_gene_product
activated	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
(	O
MAPK	B-Gene_or_gene_product
)	O
in	O
EC	B-Cell
,	O
whereas	O
an	O
inhibitor	O
of	O
MAPK	B-Gene_or_gene_product
[	O
U0126	B-Simple_chemical
,	O
1	B-Simple_chemical
,	I-Simple_chemical
4	I-Simple_chemical
-	I-Simple_chemical
diamino	I-Simple_chemical
-	I-Simple_chemical
2	I-Simple_chemical
,	I-Simple_chemical
3	I-Simple_chemical
-	I-Simple_chemical
dicyano	I-Simple_chemical
-	I-Simple_chemical
1	I-Simple_chemical
,	I-Simple_chemical
4	I-Simple_chemical
-	I-Simple_chemical
bis	I-Simple_chemical
(	I-Simple_chemical
2	I-Simple_chemical
-	I-Simple_chemical
aminophenylthio	I-Simple_chemical
)	I-Simple_chemical
butadiene	I-Simple_chemical
]	O
suppressed	O
the	O
proliferative	O
and	O
the	O
VEGF	B-Gene_or_gene_product
changes	O
but	O
not	O
the	O
pro	O
-	O
oxidant	O
effects	O
of	O
20	B-Simple_chemical
-	I-Simple_chemical
HETE	I-Simple_chemical
.	O

These	O
data	O
suggest	O
that	O
20	B-Simple_chemical
-	I-Simple_chemical
HETE	I-Simple_chemical
stimulates	O
superoxide	B-Simple_chemical
formation	O
by	O
pathways	O
other	O
than	O
apocynin	B-Simple_chemical
-	O
sensitive	O
NAD	B-Gene_or_gene_product
(	I-Gene_or_gene_product
P	I-Gene_or_gene_product
)	I-Gene_or_gene_product
H	I-Gene_or_gene_product
oxidase	I-Gene_or_gene_product
,	O
thereby	O
activating	O
MAPK	B-Gene_or_gene_product
and	O
then	O
enhancing	O
VEGF	B-Gene_or_gene_product
synthesis	O
that	O
drives	O
EC	B-Cell
proliferation	O
.	O

Thus	O
,	O
20	B-Simple_chemical
-	I-Simple_chemical
HETE	I-Simple_chemical
may	O
be	O
involved	O
in	O
the	O
regulation	O
of	O
EC	B-Cell
functions	O
,	O
such	O
as	O
angiogenesis	O
.	O

Relationship	O
of	O
cellular	B-Cell
immunity	O
,	O
cytokines	O
and	O
CRP	B-Gene_or_gene_product
with	O
clinical	O
course	O
in	O
breast	B-Cancer
cancer	I-Cancer
patients	B-Organism
with	O
endocrine	B-Anatomical_system
-	O
dependent	O
distant	O
metastases	O
treated	O
with	O
immunotherapy	O
.	O

We	O
have	O
reported	O
important	O
benefits	O
and	O
survival	O
with	O
an	O
immunotherapy	O
schedule	O
in	O
patients	B-Organism
with	O
endocrine	B-Anatomical_system
-	O
dependent	O
breast	B-Cancer
cancer	I-Cancer
and	O
distant	O
metastases	O
.	O

Here	O
clinical	O
outcome	O
is	O
updated	O
and	O
its	O
correlation	O
with	O
new	O
immunological	O
data	O
is	O
shown	O
.	O

In	O
32	O
evaluated	O
breast	B-Cancer
cancer	I-Cancer
patients	B-Organism
with	O
endocrine	B-Anatomical_system
-	O
dependent	O
distant	O
metastases	O
treated	O
with	O
a	O
new	O
immunotherapy	O
schedule	O
(	O
cyclic	O
administration	O
of	O
beta	B-Gene_or_gene_product
-	I-Gene_or_gene_product
interferon	I-Gene_or_gene_product
and	O
interleukin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
)	O
,	O
cellular	B-Cell
immunity	O
,	O
cytokines	O
and	O
CRP	B-Gene_or_gene_product
were	O
related	O
to	O
the	O
clinical	O
course	O
.	O

Estimated	O
and	O
true	O
5	O
-	O
10	O
year	O
overall	O
survival	O
rates	O
from	O
first	O
line	O
antiestrogen	O
and	O
distant	O
metastases	O
were	O
higher	O
than	O
previously	O
reported	O
in	O
a	O
similar	O
population	O
.	O

Interleukin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
administration	O
was	O
followed	O
by	O
a	O
significant	O
increase	O
in	O
total	O
lymphocytes	B-Cell
,	O
CD4	B-Gene_or_gene_product
+	O
,	O
CD8	B-Gene_or_gene_product
+	O
,	O
CD16	B-Gene_or_gene_product
+	O
56	B-Gene_or_gene_product
+	O
(	O
NK	O
)	O
cells	O
,	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
6	I-Gene_or_gene_product
,	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
12	I-Gene_or_gene_product
,	O
and	O
CRP	B-Gene_or_gene_product
(	O
from	O
P	O
<	O
0	O
.	O
04	O
to	O
P	O
<	O
0	O
.	O
000	O
)	O
but	O
no	O
change	O
in	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
10	I-Gene_or_gene_product
and	O
TGFbeta1	B-Gene_or_gene_product
during	O
clinical	O
benefit	O
.	O

During	O
progressive	O
disease	O
no	O
change	O
was	O
observed	O
in	O
the	O
former	O
parameters	O
,	O
concomitant	O
with	O
a	O
significant	O
increase	O
in	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
10	I-Gene_or_gene_product
(	O
P	O
=	O
0	O
.	O
020	O
)	O
and	O
a	O
significant	O
decrease	O
in	O
TGFbeta1	B-Gene_or_gene_product
(	O
P	O
=	O
0	O
.	O
023	O
)	O
.	O

These	O
findings	O
confirm	O
that	O
cellular	B-Cell
immunity	O
is	O
significantly	O
stimulated	O
by	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
only	O
during	O
clinical	O
benefit	O
.	O

Furthermore	O
,	O
these	O
results	O
demonstrate	O
that	O
different	O
changes	O
of	O
proinflammatory	O
cytokines	O
,	O
CRP	B-Gene_or_gene_product
and	O
inhibiting	O
factors	O
are	O
consistent	O
with	O
associated	O
clinical	O
benefit	O
or	O
with	O
disease	O
progression	O
,	O
respectively	O
.	O

Trib1	B-Gene_or_gene_product
and	O
Evi1	B-Gene_or_gene_product
cooperate	O
with	O
Hoxa	B-Gene_or_gene_product
and	O
Meis1	B-Gene_or_gene_product
in	O
myeloid	B-Cell
leukemogenesis	O
.	O

Cooperative	O
activation	O
of	O
Meis1	B-Gene_or_gene_product
and	O
Hoxa9	B-Gene_or_gene_product
perturbs	O
myeloid	B-Cell
differentiation	O
and	O
eventually	O
leads	O
myeloid	B-Cell
progenitors	I-Cell
to	O
leukemia	B-Cancer
,	O
yet	O
it	O
remains	O
to	O
be	O
clarified	O
what	O
kinds	O
of	O
subsequent	O
molecular	O
processes	O
are	O
required	O
for	O
development	O
of	O
overt	B-Cancer
leukemia	I-Cancer
.	O

To	O
understand	O
the	O
molecular	O
pathway	O
in	O
Hoxa9	B-Gene_or_gene_product
/	O
Meis1	B-Gene_or_gene_product
-	O
induced	O
leukemogenesis	O
,	O
retroviral	B-Organism
insertional	O
mutagenesis	O
was	O
applied	O
using	O
retrovirus	B-Organism
-	O
mediated	O
gene	O
transfer	O
.	O

The	O
mice	B-Organism
that	O
received	O
Hoxa9	B-Gene_or_gene_product
/	O
Meis1	B-Gene_or_gene_product
-	O
transduced	O
bone	B-Cell
marrow	I-Cell
cells	I-Cell
developed	O
acute	B-Cancer
myeloid	I-Cancer
leukemia	I-Cancer
(	O
AML	B-Cancer
)	O
,	O
and	O
Trib1	B-Gene_or_gene_product
,	O
Evi1	B-Gene_or_gene_product
,	O
Ahi1	B-Gene_or_gene_product
,	O
Raralpha	B-Gene_or_gene_product
,	O
Pitpnb	B-Gene_or_gene_product
,	O
and	O
AK039950	B-Gene_or_gene_product
were	O
identified	O
as	O
candidate	O
cooperative	O
genes	O
located	O
near	O
common	O
retroviral	O
integration	O
sites	O
.	O

Trib1	B-Gene_or_gene_product
and	O
Evi1	B-Gene_or_gene_product
were	O
up	O
-	O
regulated	O
due	O
to	O
retroviral	B-Organism
insertions	O
,	O
and	O
coexpression	O
of	O
these	O
genes	O
significantly	O
accelerated	O
the	O
onset	O
of	O
Hoxa9	B-Gene_or_gene_product
/	O
Meis1	B-Gene_or_gene_product
-	O
induced	O
AML	B-Cancer
,	O
suggesting	O
that	O
Trib1	B-Gene_or_gene_product
and	O
Evi1	B-Gene_or_gene_product
are	O
the	O
key	O
collaborators	O
.	O

Furthermore	O
,	O
Trib1	B-Gene_or_gene_product
by	O
itself	O
is	O
a	O
novel	O
myeloid	B-Cell
oncogene	O
,	O
enhancing	O
phosphorylation	O
of	O
ERK	B-Gene_or_gene_product
,	O
resulting	O
in	O
inhibition	O
of	O
apoptosis	O
.	O

These	O
results	O
demonstrate	O
the	O
importance	O
of	O
specific	O
oncogene	O
interaction	O
in	O
myeloid	B-Cell
leukemogenesis	O
.	O

Pro	O
/	O
antioxidant	O
status	O
and	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
transcription	O
factor	O
in	O
murine	B-Organism
skin	B-Organ
following	O
topical	O
exposure	O
to	O
cumene	B-Simple_chemical
hydroperoxide	I-Simple_chemical
.	O

Organic	B-Simple_chemical
peroxides	I-Simple_chemical
,	O
widely	O
used	O
in	O
the	O
chemical	O
and	O
pharmaceutical	O
industries	O
,	O
can	O
act	O
as	O
skin	B-Cancer
tumor	I-Cancer
promoters	O
and	O
cause	O
epidermal	B-Pathological_formation
hyperplasia	I-Pathological_formation
.	O

They	O
are	O
also	O
known	O
to	O
trigger	O
free	B-Simple_chemical
radical	I-Simple_chemical
generation	O
.	O

The	O
present	O
study	O
evaluated	O
the	O
effect	O
of	O
cumene	B-Simple_chemical
hydroperoxide	I-Simple_chemical
(	O
Cum	B-Simple_chemical
-	I-Simple_chemical
OOH	I-Simple_chemical
)	O
on	O
the	O
induction	O
of	O
activator	B-Gene_or_gene_product
protein	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
,	O
which	O
is	O
linked	O
to	O
the	O
expression	O
of	O
genes	O
regulating	O
cell	B-Cell
proliferation	O
,	O
growth	O
and	O
transformation	O
.	O

Previously	O
,	O
we	O
reported	O
that	O
topical	O
exposure	O
to	O
Cum	B-Simple_chemical
-	I-Simple_chemical
OOH	I-Simple_chemical
caused	O
formation	O
of	O
free	B-Simple_chemical
radicals	I-Simple_chemical
and	O
oxidative	O
stress	O
in	O
the	O
skin	B-Organ
of	O
vitamin	B-Simple_chemical
E	I-Simple_chemical
-	O
deficient	O
mice	B-Organism
.	O

The	O
present	O
study	O
used	O
JB6	O
P	O
+	O
mouse	B-Organism
epidermal	O
cells	O
and	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
luciferase	B-Gene_or_gene_product
reporter	O
transgenic	O
mice	B-Organism
to	O
identify	O
whether	O
exposure	O
to	O
Cum	B-Simple_chemical
-	I-Simple_chemical
OOH	I-Simple_chemical
caused	O
activation	O
of	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
oxidative	O
stress	O
,	O
depletion	O
of	O
antioxidants	O
and	O
tumor	B-Cancer
formation	O
during	O
two	O
-	O
stage	O
carcinogenesis	O
.	O

In	O
vitro	O
studies	O
found	O
that	O
exposure	O
to	O
Cum	B-Simple_chemical
-	I-Simple_chemical
OOH	I-Simple_chemical
reduced	O
the	O
level	O
of	O
glutathione	B-Simple_chemical
(	O
GSH	B-Simple_chemical
)	O
in	O
mouse	B-Organism
epidermal	B-Cell
cells	I-Cell
(	O
JB6	B-Cell
P	I-Cell
+	I-Cell
)	O
and	O
caused	O
the	O
induction	O
of	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

Mice	B-Organism
primed	O
with	O
dimethyl	B-Simple_chemical
-	I-Simple_chemical
benz	I-Simple_chemical
[	I-Simple_chemical
a	I-Simple_chemical
]	I-Simple_chemical
anthracene	I-Simple_chemical
(	O
DMBA	B-Simple_chemical
)	O
were	O
topically	O
exposed	O
to	O
Cum	B-Simple_chemical
-	I-Simple_chemical
OOH	I-Simple_chemical
(	O
82	O
.	O
6	O
micromol	O
)	O
or	O
the	O
positive	O
control	O
,	O
12	B-Simple_chemical
-	I-Simple_chemical
O	I-Simple_chemical
-	I-Simple_chemical
tetradecanoylphorbol	I-Simple_chemical
-	I-Simple_chemical
13	I-Simple_chemical
-	I-Simple_chemical
acetate	I-Simple_chemical
(	O
TPA	B-Simple_chemical
,	O
17	O
nmol	O
)	O
,	O
twice	O
weekly	O
for	O
29	O
weeks	O
.	O

Activation	O
of	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
in	O
skin	B-Organ
was	O
detected	O
as	O
early	O
as	O
2	O
weeks	O
following	O
Cum	B-Simple_chemical
-	I-Simple_chemical
OOH	I-Simple_chemical
or	O
TPA	B-Simple_chemical
exposure	O
.	O

No	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
expression	O
was	O
found	O
19	O
weeks	O
after	O
initiation	O
.	O

Papilloma	B-Pathological_formation
formation	O
was	O
observed	O
in	O
both	O
the	O
DMBA	B-Simple_chemical
-	O
TPA	B-Simple_chemical
-	O
and	O
DMBA	B-Simple_chemical
-	O
Cum	B-Simple_chemical
-	I-Simple_chemical
OOH	I-Simple_chemical
-	O
exposed	O
animals	B-Organism
,	O
whereas	O
skin	B-Cancer
carcinomas	I-Cancer
were	O
found	O
only	O
in	O
the	O
DMBA	B-Simple_chemical
-	O
Cum	B-Simple_chemical
-	I-Simple_chemical
OOH	I-Simple_chemical
-	O
treated	O
mice	B-Organism
.	O

A	O
greater	O
accumulation	O
of	O
peroxidative	O
products	O
(	O
thiobarbituric	B-Simple_chemical
acid	I-Simple_chemical
-	O
reactive	O
substances	O
)	O
,	O
inflammation	O
and	O
decreased	O
levels	O
of	O
GSH	B-Simple_chemical
and	O
total	O
antioxidant	O
reserves	O
were	O
also	O
observed	O
in	O
the	O
skin	B-Organ
of	O
DMBA	B-Simple_chemical
-	O
Cum	B-Simple_chemical
-	I-Simple_chemical
OOH	I-Simple_chemical
-	O
exposed	O
mice	B-Organism
.	O

These	O
results	O
suggest	O
that	O
Cum	B-Simple_chemical
-	I-Simple_chemical
OOH	I-Simple_chemical
-	O
induced	O
carcinogenesis	O
is	O
accompanied	O
by	O
increased	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
activation	O
and	O
changes	O
in	O
antioxidant	O
status	O
.	O

Hyperforin	B-Simple_chemical
blocks	O
neutrophil	B-Cell
activation	O
of	O
matrix	B-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
,	O
motility	O
and	O
recruitment	O
,	O
and	O
restrains	O
inflammation	O
-	O
triggered	O
angiogenesis	O
and	O
lung	B-Organ
fibrosis	O
.	O

Hyperforin	B-Simple_chemical
(	O
Hyp	B-Simple_chemical
)	O
,	O
a	O
polyphenol	B-Simple_chemical
-	O
derivative	O
of	O
St	B-Organism
.	I-Organism

John	B-Organism
'	I-Organism
s	I-Organism
wort	I-Organism
(	O
Hypericum	B-Organism
perforatum	I-Organism
)	O
,	O
has	O
emerged	O
as	O
key	O
player	O
not	O
only	O
in	O
the	O
antidepressant	O
activity	O
of	O
the	O
plant	O
but	O
also	O
as	O
an	O
inhibitor	O
of	O
bacteria	O
lymphocyte	B-Cell
and	O
tumor	B-Cell
cell	I-Cell
proliferation	O
,	O
and	O
matrix	B-Gene_or_gene_product
proteinases	I-Gene_or_gene_product
.	O

We	O
tested	O
whether	O
as	O
well	O
as	O
inhibiting	O
leukocyte	B-Gene_or_gene_product
elastase	I-Gene_or_gene_product
(	O
LE	B-Gene_or_gene_product
)	O
activity	O
,	O
Hyp	B-Simple_chemical
might	O
be	O
effective	O
in	O
containing	O
both	O
polymorphonuclear	B-Cell
neutrophil	I-Cell
(	I-Cell
PMN	I-Cell
)	I-Cell
leukocyte	I-Cell
recruitment	O
and	O
unfavorable	O
eventual	O
tissue	B-Tissue
responses	O
.	O

The	O
results	O
show	O
that	O
,	O
without	O
affecting	O
in	O
vitro	O
human	B-Organism
PMN	B-Cell
viability	O
and	O
chemokine	O
-	O
receptor	O
expression	O
,	O
Hyp	B-Simple_chemical
(	O
as	O
stable	O
dicyclohexylammonium	B-Simple_chemical
salt	I-Simple_chemical
)	O
was	O
able	O
to	O
inhibit	O
in	O
a	O
dose	O
-	O
dependent	O
manner	O
their	O
chemotaxis	O
and	O
chemoinvasion	O
(	O
IC50	O
=	O
1	O
microM	O
for	O
both	O
)	O
;	O
this	O
effect	O
was	O
associated	O
with	O
a	O
reduced	O
expression	O
of	O
the	O
adhesion	O
molecule	O
CD11b	B-Gene_or_gene_product
by	O
formyl	B-Simple_chemical
-	I-Simple_chemical
Met	I-Simple_chemical
-	I-Simple_chemical
Leu	I-Simple_chemical
-	I-Simple_chemical
Phe	I-Simple_chemical
-	O
stimulated	O
neutrophils	B-Cell
and	O
block	O
of	O
LE	B-Gene_or_gene_product
-	O
triggered	O
activation	O
of	O
the	O
gelatinase	B-Gene_or_gene_product
matrix	I-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
.	O

PMN	B-Cell
-	O
triggered	O
angiogenesis	O
is	O
also	O
blocked	O
by	O
both	O
local	O
injection	O
and	O
daily	O
i	O
.	O
p	O
.	O
administration	O
of	O
the	O
Hyp	B-Simple_chemical
salt	O
in	O
an	O
interleukin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
-	O
induced	O
murine	B-Organism
model	O
.	O

Furthermore	O
,	O
i	O
.	O
p	O
.	O
treatment	O
with	O
Hyp	B-Simple_chemical
reduces	O
acute	O
PMN	B-Cell
recruitment	O
and	O
enhances	O
resolution	O
in	O
a	O
pulmonary	B-Organ
bleomycin	B-Simple_chemical
-	O
induced	O
inflammation	O
model	O
,	O
significantly	O
reducing	O
consequent	O
fibrosis	O
.	O

These	O
results	O
indicate	O
that	O
Hyp	B-Simple_chemical
is	O
a	O
powerful	O
anti	O
-	O
inflammatory	O
compound	O
with	O
therapeutic	O
potential	O
,	O
and	O
they	O
elucidate	O
mechanistic	O
keys	O
.	O

Impaired	O
apoptosis	O
of	O
pulmonary	B-Cell
endothelial	I-Cell
cells	I-Cell
is	O
associated	O
with	O
intimal	B-Tissue
proliferation	O
and	O
irreversibility	O
of	O
pulmonary	B-Organ
hypertension	O
in	O
congenital	O
heart	B-Organ
disease	O
.	O

OBJECTIVES	O
:	O
This	O
study	O
sought	O
to	O
assess	O
the	O
cellular	B-Cell
and	O
histologic	O
basis	O
of	O
irreversible	O
pulmonary	B-Organ
hypertension	O
(	O
PHT	O
)	O
in	O
the	O
clinical	O
setting	O
of	O
congenital	O
heart	B-Organ
disease	O
(	O
CHD	O
)	O
.	O

BACKGROUND	O
:	O
Although	O
many	O
children	B-Organism
with	O
CHD	O
develop	O
pulmonary	B-Multi-tissue_structure
vascular	I-Multi-tissue_structure
disease	O
,	O
it	O
is	O
unclear	O
why	O
this	O
complication	O
is	O
reversible	O
after	O
complete	O
repair	O
in	O
some	O
cases	O
but	O
irreversible	O
in	O
others	O
.	O

Because	O
failure	O
of	O
endothelial	B-Cell
cell	I-Cell
apoptosis	O
might	O
lead	O
to	O
intimal	B-Tissue
proliferation	O
and	O
lack	O
of	O
reversibility	O
of	O
PHT	O
,	O
we	O
investigated	O
this	O
and	O
other	O
key	O
markers	O
of	O
vasoactivity	O
and	O
angiogenesis	O
in	O
subjects	O
with	O
PHT	O
and	O
CHD	O
.	O

METHODS	O
:	O
We	O
assessed	O
antiapoptotic	O
and	O
proapoptotic	O
markers	O
in	O
vascular	B-Cell
and	O
perivascular	B-Cell
cells	I-Cell
in	O
lung	B-Multi-tissue_structure
biopsy	I-Multi-tissue_structure
samples	I-Multi-tissue_structure
from	O
18	O
patients	B-Organism
with	O
CHD	O
,	O
7	O
with	O
reversible	O
and	O
11	O
with	O
irreversible	O
PHT	O
,	O
and	O
6	O
control	O
patients	B-Organism
.	O

Immunostaining	O
for	O
endothelial	B-Gene_or_gene_product
nitric	I-Gene_or_gene_product
oxide	I-Gene_or_gene_product
synthase	I-Gene_or_gene_product
,	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
,	O
and	O
CD34	B-Gene_or_gene_product
(	O
markers	O
of	O
vasoactivity	O
and	O
neoangiogenesis	O
)	O
was	O
also	O
performed	O
.	O

RESULTS	O
:	O
The	O
antiapoptotic	O
protein	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
was	O
highly	O
expressed	O
by	O
pulmonary	B-Cell
endothelial	I-Cell
cells	I-Cell
in	O
all	O
cases	O
of	O
irreversible	O
PHT	O
but	O
in	O
no	O
cases	O
of	O
reversible	O
PHT	O
,	O
nor	O
in	O
control	O
patients	B-Organism
(	O
p	O
less	O
than	O
0	O
.	O
001	O
)	O
.	O

Intimal	B-Tissue
proliferation	O
was	O
present	O
in	O
10	O
of	O
11	O
irreversible	O
PHT	O
cases	O
,	O
but	O
never	O
observed	O
in	O
reversible	O
PHT	O
(	O
p	O
less	O
than	O
0	O
.	O
001	O
)	O
.	O

Similarly	O
,	O
perivascular	B-Cell
inflammatory	I-Cell
T	I-Cell
-	I-Cell
cells	I-Cell
expressed	O
more	O
antiapoptotic	O
proteins	O
in	O
irreversible	O
PHT	O
(	O
p	O
less	O
than	O
0	O
.	O
01	O
)	O
.	O

Irreversible	O
PHT	O
cases	O
were	O
also	O
more	O
likely	O
to	O
show	O
compensatory	O
upregulation	O
of	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
and	O
new	O
small	O
vessel	B-Multi-tissue_structure
formation	O
at	O
the	O
sites	O
of	O
native	O
vessel	B-Multi-tissue_structure
stenosis	O
or	O
occlusion	O
(	O
p	O
less	O
than	O
0	O
.	O
001	O
)	O
.	O

CONCLUSIONS	O
:	O
Irreversible	O
PHT	O
is	O
strongly	O
associated	O
with	O
impaired	O
endothelial	B-Cell
cell	I-Cell
apoptosis	O
and	O
antiapoptotic	O
signaling	O
from	O
perivascular	B-Cell
inflammatory	I-Cell
cells	I-Cell
.	O

These	O
changes	O
are	O
associated	O
with	O
intimal	B-Tissue
proliferation	O
and	O
vessel	B-Multi-tissue_structure
narrowing	O
,	O
and	O
thereby	O
may	O
contribute	O
to	O
clinical	O
outcomes	O
associated	O
with	O
pulmonary	B-Organ
hypertension	O
.	O

Inhibition	O
of	O
Dll4	B-Gene_or_gene_product
-	O
mediated	O
signaling	O
induces	O
proliferation	O
of	O
immature	O
vessels	B-Multi-tissue_structure
and	O
results	O
in	O
poor	O
tissue	B-Tissue
perfusion	O
.	O

Vascular	B-Multi-tissue_structure
development	O
is	O
dependent	O
on	O
various	O
growth	O
factors	O
and	O
certain	O
modifiers	O
critical	O
for	O
providing	O
arterial	B-Multi-tissue_structure
or	O
venous	B-Multi-tissue_structure
identity	O
,	O
interaction	O
with	O
the	O
surrounding	O
stroma	B-Tissue
and	O
tissues	B-Tissue
,	O
hierarchic	B-Tissue
network	I-Tissue
formation	O
,	O
and	O
recruitment	O
of	O
pericytes	B-Cell
.	O

Notch	B-Gene_or_gene_product
receptors	O
and	O
ligands	O
(	O
Jagged	B-Gene_or_gene_product
and	O
Delta	B-Gene_or_gene_product
-	O
like	O
)	O
play	O
a	O
critical	O
role	O
in	O
this	O
process	O
in	O
addition	O
to	O
VEGF	B-Gene_or_gene_product
.	O

Dll4	B-Gene_or_gene_product
is	O
one	O
of	O
the	O
Notch	B-Gene_or_gene_product
ligands	O
that	O
regulates	O
arterial	B-Multi-tissue_structure
specification	O
and	O
maturation	O
events	O
.	O

In	O
the	O
current	O
study	O
,	O
we	O
have	O
shown	O
that	O
loss	O
of	O
function	O
by	O
either	O
targeted	O
allele	O
deletion	O
or	O
use	O
of	O
a	O
soluble	O
form	O
of	O
Dll4	B-Gene_or_gene_product
extracellular	B-Immaterial_anatomical_entity
domain	O
leads	O
to	O
inhibition	O
of	O
Notch	B-Gene_or_gene_product
signaling	O
,	O
resulting	O
in	O
increased	O
vascular	B-Multi-tissue_structure
proliferation	O
but	O
defective	O
maturation	O
.	O

Newly	O
forming	O
vessels	B-Multi-tissue_structure
have	O
thin	O
caliber	B-Immaterial_anatomical_entity
,	O
a	O
markedly	O
reduced	O
vessel	B-Immaterial_anatomical_entity
lumen	I-Immaterial_anatomical_entity
,	O
markedly	O
reduced	O
pericyte	B-Cell
recruitment	O
,	O
and	O
deficient	O
vascular	B-Multi-tissue_structure
perfusion	O
.	O

sDll4	B-Gene_or_gene_product
similarly	O
induced	O
defective	O
vascular	B-Multi-tissue_structure
response	O
in	O
tumor	B-Cancer
implants	O
leading	O
to	O
reduced	O
tumor	B-Cancer
growth	O
.	O

Interference	O
with	O
Dll4	B-Gene_or_gene_product
-	O
Notch	B-Gene_or_gene_product
signaling	O
may	O
be	O
particularly	O
desirable	O
in	O
tumors	B-Cancer
that	O
have	O
highly	O
induced	O
Dll4	B-Gene_or_gene_product
-	O
Notch	B-Gene_or_gene_product
pathway	O
.	O

SWAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
70	I-Gene_or_gene_product
is	O
required	O
for	O
oncogenic	O
transformation	O
by	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Src	I-Gene_or_gene_product
in	O
mouse	B-Organism
embryo	B-Cell
fibroblasts	I-Cell
.	O

SWAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
70	I-Gene_or_gene_product
is	O
a	O
phosphatidylinositol	B-Simple_chemical
trisphosphate	I-Simple_chemical
(	O
PtdIns	B-Simple_chemical
(	I-Simple_chemical
3	I-Simple_chemical
,	I-Simple_chemical
4	I-Simple_chemical
,	I-Simple_chemical
5	I-Simple_chemical
)	I-Simple_chemical
P	I-Simple_chemical
(	I-Simple_chemical
3	I-Simple_chemical
)	I-Simple_chemical
)	O
binding	O
protein	O
,	O
which	O
acts	O
in	O
F	B-Gene_or_gene_product
-	I-Gene_or_gene_product
actin	I-Gene_or_gene_product
rearrangement	O
.	O

The	O
role	O
of	O
SWAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
70	I-Gene_or_gene_product
in	O
oncogenic	O
transformation	O
of	O
mouse	B-Organism
embryo	B-Cell
fibroblasts	I-Cell
(	O
MEFs	B-Cell
)	O
by	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Src	I-Gene_or_gene_product
was	O
examined	O
by	O
use	O
of	O
MEFs	B-Cell
defective	O
in	O
SWAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
70	I-Gene_or_gene_product
.	O

v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Src	I-Gene_or_gene_product
morphologically	O
transformed	O
MEFs	B-Cell
lacking	O
SWAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
70	I-Gene_or_gene_product
,	O
but	O
growth	O
of	O
the	O
transformed	O
cells	B-Cell
in	O
culture	O
was	O
slower	O
than	O
that	O
of	O
cells	B-Cell
supplemented	O
with	O
exogenous	O
SWAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
70	I-Gene_or_gene_product
.	O

The	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Src	I-Gene_or_gene_product
-	O
transformed	O
MEFs	B-Cell
deficient	O
in	O
SWAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
70	I-Gene_or_gene_product
were	O
unable	O
to	O
grow	O
in	O
soft	O
agar	O
while	O
those	O
expressing	O
SWAP70	B-Gene_or_gene_product
readily	O
formed	O
colonies	B-Cell
,	O
suggesting	O
that	O
SWAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
70	I-Gene_or_gene_product
is	O
required	O
for	O
anchorage	O
independent	O
growth	O
of	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Src	I-Gene_or_gene_product
transformed	O
MEFs	B-Cell
.	O

When	O
transplanted	O
in	O
nude	B-Organism
mice	I-Organism
,	O
tumors	B-Cancer
formed	O
by	O
the	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Src	I-Gene_or_gene_product
transformed	O
SWAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
70	I-Gene_or_gene_product
(	O
-	O
/	O
-	O
)	O
MEFs	O
were	O
smaller	O
than	O
those	O
formed	O
by	O
cells	B-Cell
expressing	O
exogenous	O
SWAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
70	I-Gene_or_gene_product
.	O

These	O
results	O
suggest	O
that	O
SWAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
70	I-Gene_or_gene_product
may	O
be	O
required	O
for	O
oncogenic	O
transformation	O
and	O
contributes	O
to	O
cell	B-Cell
growth	O
in	O
MEFs	B-Cell
transformed	O
by	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Src	I-Gene_or_gene_product
.	O

Tsg101	B-Gene_or_gene_product
is	O
upregulated	O
in	O
a	O
subset	O
of	O
invasive	O
human	B-Organism
breast	B-Cancer
cancers	I-Cancer
and	O
its	O
targeted	O
overexpression	O
in	O
transgenic	B-Organism
mice	I-Organism
reveals	O
weak	O
oncogenic	O
properties	O
for	O
mammary	B-Cancer
cancer	I-Cancer
initiation	O
.	O

Previous	O
studies	O
reported	O
that	O
the	O
Tumor	B-Gene_or_gene_product
Susceptibility	I-Gene_or_gene_product
Gene	I-Gene_or_gene_product
101	I-Gene_or_gene_product
(	O
TSG101	B-Gene_or_gene_product
)	O
is	O
upregulated	O
in	O
selected	O
human	B-Organism
malignancies	B-Cancer
,	O
and	O
the	O
expression	O
of	O
exogenous	O
Tsg101	B-Gene_or_gene_product
was	O
suggested	O
to	O
transform	O
immortalized	O
fibroblasts	B-Cell
in	O
culture	O
.	O

To	O
date	O
,	O
the	O
potential	O
oncogenic	O
properties	O
of	O
Tsg101	B-Gene_or_gene_product
have	O
not	O
been	O
examined	O
in	O
vivo	O
owing	O
to	O
the	O
lack	O
of	O
appropriate	O
model	O
systems	O
.	O

In	O
this	O
study	O
,	O
we	O
show	O
that	O
Tsg101	B-Gene_or_gene_product
is	O
highly	O
expressed	O
in	O
a	O
subset	O
of	O
invasive	O
human	B-Organism
breast	B-Cancer
cancers	I-Cancer
.	O

Based	O
on	O
this	O
observation	O
,	O
we	O
generated	O
the	O
first	O
transgenic	O
mouse	B-Organism
model	O
with	O
a	O
targeted	O
overexpression	O
of	O
Tsg101	B-Gene_or_gene_product
in	O
the	O
developing	O
mammary	B-Organ
gland	I-Organ
to	O
test	O
whether	O
exogenous	O
Tsg101	B-Gene_or_gene_product
is	O
capable	O
of	O
initiating	O
tumorigenesis	O
.	O

Normal	O
functionality	O
of	O
exogenous	O
Tsg101	B-Gene_or_gene_product
was	O
tested	O
by	O
rescuing	O
the	O
survival	O
of	O
Tsg101	B-Gene_or_gene_product
-	O
deficient	O
mammary	B-Cell
epithelial	I-Cell
cells	I-Cell
in	O
conditional	O
knockout	O
mice	B-Organism
.	O

The	O
overexpression	O
of	O
Tsg101	B-Gene_or_gene_product
resulted	O
in	O
increased	O
phosphorylation	O
of	O
the	O
epidermal	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
and	O
downstream	O
activation	O
of	O
MAP	B-Gene_or_gene_product
kinases	I-Gene_or_gene_product
.	O

Despite	O
an	O
increase	O
in	O
the	O
activation	O
of	O
these	O
signal	O
transducers	O
,	O
the	O
mammary	B-Organ
gland	I-Organ
of	O
females	O
expressing	O
exogenous	O
Tsg101	B-Gene_or_gene_product
developed	O
normally	O
throughout	O
the	O
reproductive	O
cycle	O
.	O

In	O
aging	O
females	O
,	O
the	O
overexpression	O
of	O
Tsg101	B-Gene_or_gene_product
seemed	O
to	O
increase	O
the	O
susceptibility	O
of	O
mammary	B-Tissue
epithelia	I-Tissue
toward	O
malignant	O
transformation	O
.	O

However	O
,	O
owing	O
to	O
the	O
long	O
latency	O
of	O
tumor	B-Cancer
formation	O
and	O
the	O
sporadic	O
occurrence	O
of	O
bona	O
fide	O
mammary	B-Cancer
cancers	I-Cancer
,	O
we	O
conclude	O
that	O
the	O
Tsg101	B-Gene_or_gene_product
protein	O
has	O
only	O
weak	O
oncogenic	O
properties	O
.	O

Instead	O
of	O
cancer	B-Cancer
initiation	O
,	O
it	O
is	O
therefore	O
likely	O
that	O
Tsg101	B-Gene_or_gene_product
plays	O
a	O
more	O
predominant	O
role	O
in	O
the	O
progression	O
of	O
a	O
subset	O
of	O
spontaneously	O
arising	O
breast	B-Cancer
cancers	I-Cancer
.	O

Expression	O
of	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
xL	I-Gene_or_gene_product
,	O
Bax	B-Gene_or_gene_product
,	O
and	O
p53	B-Gene_or_gene_product
in	O
primary	B-Cancer
tumors	I-Cancer
and	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastases	O
in	O
oral	B-Cancer
squamous	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
.	O

Disturbances	O
in	O
expression	O
of	O
apoptosis	O
-	O
associated	O
proteins	O
take	O
part	O
in	O
the	O
development	O
and	O
progression	O
of	O
many	O
human	B-Organism
malignancies	B-Cancer
.	O

The	O
aim	O
of	O
this	O
study	O
was	O
the	O
assessment	O
of	O
correlations	O
among	O
proteins	O
involved	O
in	O
apoptosis	O
-	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
xL	I-Gene_or_gene_product
,	O
Bax	B-Gene_or_gene_product
,	O
and	O
p53	B-Gene_or_gene_product
-	O
as	O
well	O
as	O
relationships	O
of	O
these	O
proteins	O
with	O
selected	O
clinicopathological	O
features	O
in	O
oral	B-Cancer
squamous	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
.	O

Consequently	O
,	O
we	O
examined	O
by	O
immunohistochemistry	O
,	O
using	O
the	O
avidin	B-Gene_or_gene_product
-	O
biotin	B-Simple_chemical
-	O
peroxidase	B-Gene_or_gene_product
method	O
,	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
xL	I-Gene_or_gene_product
,	O
Bax	B-Gene_or_gene_product
,	O
and	O
p53	B-Gene_or_gene_product
expression	O
in	O
56	O
samples	O
of	O
primary	O
oral	B-Cancer
squamous	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
and	O
in	O
22	O
matched	O
pairs	O
of	O
primary	O
and	O
metastatic	B-Cancer
tumors	I-Cancer
.	O

The	O
evaluation	O
of	O
immunostaining	O
of	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
xL	I-Gene_or_gene_product
,	O
Bax	B-Gene_or_gene_product
,	O
and	O
p53	B-Gene_or_gene_product
was	O
analyzed	O
in	O
10	O
different	O
tumor	B-Cancer
fields	O
,	O
and	O
the	O
mean	O
percentage	O
of	O
tumor	B-Cell
cells	I-Cell
with	O
positive	O
staining	O
was	O
evaluated	O
.	O

The	O
significance	O
of	O
the	O
associations	O
was	O
determined	O
using	O
Spearman	O
correlation	O
analysis	O
and	O
the	O
chi	O
-	O
square	O
test	O
.	O

We	O
found	O
positive	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
xL	I-Gene_or_gene_product
,	O
Bax	B-Gene_or_gene_product
,	O
and	O
p53	B-Gene_or_gene_product
immunostaining	O
in	O
44	O
.	O
6	O
%	O
,	O
28	O
.	O
6	O
%	O
,	O
and	O
58	O
.	O
9	O
%	O
of	O
the	O
studied	O
primary	O
tumors	B-Cancer
and	O
in	O
63	O
.	O
6	O
%	O
,	O
45	O
.	O
5	O
%	O
,	O
and	O
72	O
.	O
7	O
%	O
of	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastases	O
,	O
respectively	O
.	O

Analysis	O
of	O
associations	O
among	O
studied	O
proteins	O
revealed	O
positive	O
correlation	O
between	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
xL	I-Gene_or_gene_product
and	O
Bax	B-Gene_or_gene_product
in	O
primary	O
tumors	B-Cancer
(	O
P	O
<	O
0	O
.	O
03	O
,	O
r	O
=	O
0	O
.	O
307	O
)	O
.	O

Statistically	O
significant	O
relationship	O
between	O
p53	B-Gene_or_gene_product
expression	O
in	O
primary	O
oral	B-Organism_subdivision
cancers	B-Cancer
and	O
its	O
expression	O
in	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastases	O
(	O
P	O
<	O
0	O
.	O
02	O
)	O
as	O
well	O
as	O
increased	O
expression	O
of	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
xL	I-Gene_or_gene_product
,	O
Bax	B-Gene_or_gene_product
,	O
and	O
p53	B-Gene_or_gene_product
in	O
metastatic	O
sites	O
compared	O
with	O
primary	O
tumors	B-Cancer
could	O
indicate	O
an	O
association	O
of	O
these	O
proteins	O
with	O
oral	B-Cancer
cancer	I-Cancer
progression	O
and	O
development	O
of	O
metastases	O
.	O

Moreover	O
,	O
we	O
suppose	O
that	O
knowledge	O
about	O
heterogeneity	O
between	O
primary	O
and	O
metastatic	O
tumor	B-Cancer
might	O
help	O
to	O
understand	O
mechanisms	O
of	O
oral	B-Cancer
cancer	I-Cancer
progression	O
.	O

An	O
antibody	B-Gene_or_gene_product
directed	I-Gene_or_gene_product
against	I-Gene_or_gene_product
PDGF	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
enhances	O
the	O
antitumor	B-Cancer
and	O
the	O
anti	O
-	O
angiogenic	O
activities	O
of	O
an	O
anti	B-Gene_or_gene_product
-	I-Gene_or_gene_product
VEGF	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
2	I-Gene_or_gene_product
antibody	I-Gene_or_gene_product
.	O

Platelet	B-Gene_or_gene_product
-	I-Gene_or_gene_product
derived	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
PDGF	B-Gene_or_gene_product
)	O
and	O
its	O
receptors	O
(	O
PDGFR	B-Gene_or_gene_product
)	O
play	O
important	O
roles	O
in	O
tumorigenesis	O
through	O
stimulating	O
tumor	B-Cancer
growth	O
and	O
promoting	O
angiogenesis	O
via	O
enhancing	O
pericyte	B-Cell
recruitment	O
and	O
vessel	B-Multi-tissue_structure
maturation	O
.	O

Here	O
we	O
produced	O
a	O
neutralizing	O
antibody	O
,	O
1B3	B-Simple_chemical
,	O
directed	O
against	O
mouse	B-Organism
PDGFRbeta	B-Gene_or_gene_product
.	O

1B3	B-Simple_chemical
binds	O
to	O
PDGFRbeta	B-Gene_or_gene_product
with	O
high	O
affinity	O
(	O
9x10	O
(	O
-	O
11	O
)	O
M	O
)	O
and	O
blocks	O
PDGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
BB	I-Gene_or_gene_product
from	O
binding	O
to	O
the	O
receptor	O
with	O
an	O
IC	O
(	O
50	O
)	O
of	O
approximately	O
1	O
.	O
2	O
nM	O
.	O

The	O
antibody	O
also	O
blocks	O
ligand	O
-	O
stimulated	O
activation	O
of	O
PDGFRbeta	B-Gene_or_gene_product
and	O
downstream	O
signaling	O
molecules	O
,	O
including	O
Akt	B-Gene_or_gene_product
and	O
MAPK	B-Gene_or_gene_product
p42	I-Gene_or_gene_product
/	I-Gene_or_gene_product
44	I-Gene_or_gene_product
,	O
in	O
tumor	B-Cell
cells	I-Cell
.	O

In	O
animal	O
studies	O
,	O
1B3	B-Simple_chemical
significantly	O
enhanced	O
the	O
antitumor	B-Cancer
and	O
the	O
anti	O
-	O
angiogenic	O
activities	O
of	O
DC101	B-Simple_chemical
,	O
an	O
antibody	O
directed	O
against	O
mouse	B-Organism
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
in	O
a	O
pancreatic	B-Cancer
(	O
BxPC	B-Cancer
-	I-Cancer
3	I-Cancer
)	O
and	O
a	O
non	B-Cancer
-	I-Cancer
small	I-Cancer
cell	I-Cancer
lung	I-Cancer
(	I-Cancer
NCI	I-Cancer
-	I-Cancer
H460	I-Cancer
)	I-Cancer
tumor	I-Cancer
xenograft	I-Cancer
models	O
.	O

Treatment	O
with	O
the	O
combination	O
of	O
1B3	B-Simple_chemical
and	O
DC101	B-Simple_chemical
in	O
BxPC	B-Cancer
-	I-Cancer
3	I-Cancer
xenograft	I-Cancer
-	O
bearing	O
mice	B-Organism
resulted	O
in	O
tumor	B-Cancer
regression	O
in	O
58	O
%	O
of	O
mice	B-Organism
compared	O
to	O
that	O
in	O
18	O
%	O
of	O
mice	B-Organism
treated	O
with	O
DC101	B-Simple_chemical
alone	O
.	O

Taken	O
together	O
,	O
these	O
results	O
lend	O
great	O
support	O
to	O
use	O
PDGFRbeta	B-Gene_or_gene_product
antagonists	I-Gene_or_gene_product
in	O
combinations	O
with	O
other	O
antitumor	B-Cancer
and	O
/	O
or	O
anti	O
-	O
angiogenic	O
agents	O
in	O
the	O
treatment	O
of	O
a	O
variety	O
of	O
cancers	B-Cancer
.	O

In	O
vivo	O
p53	B-Gene_or_gene_product
response	O
and	O
immune	O
reaction	O
underlie	O
highly	O
effective	O
low	O
-	O
dose	O
radiotherapy	O
in	O
follicular	B-Cancer
lymphoma	I-Cancer
.	O

Very	O
low	O
-	O
dose	O
irradiation	O
(	O
2	O
x	O
2	O
Gy	O
)	O
is	O
a	O
new	O
,	O
effective	O
,	O
and	O
safe	O
local	O
treatment	O
for	O
follicular	B-Cancer
lymphoma	I-Cancer
.	O

To	O
understand	O
the	O
biologic	O
mechanisms	O
of	O
this	O
extremely	O
effective	O
response	O
,	O
we	O
compared	O
by	O
microarray	O
the	O
gene	O
-	O
expression	O
profile	O
of	O
patients	B-Organism
'	O
biopsies	B-Cancer
taken	O
before	O
and	O
after	O
radiation	O
.	O

In	O
all	O
patients	B-Organism
,	O
a	O
major	O
and	O
consistent	O
induction	O
of	O
p53	B-Gene_or_gene_product
target	O
genes	O
was	O
seen	O
.	O

p53	B-Gene_or_gene_product
targets	O
involved	O
in	O
cell	B-Cell
-	O
cycle	O
arrest	O
and	O
apoptosis	O
showed	O
the	O
same	O
mode	O
of	O
regulation	O
,	O
indicating	O
that	O
,	O
in	O
vivo	O
,	O
both	O
are	O
activated	O
simultaneously	O
.	O

p53	B-Gene_or_gene_product
up	O
-	O
regulation	O
and	O
p53	B-Gene_or_gene_product
-	O
mediated	O
proliferation	O
arrest	O
and	O
apoptosis	O
were	O
substantiated	O
using	O
immunohistochemistry	O
,	O
with	O
activation	O
of	O
both	O
the	O
intrinsic	O
and	O
the	O
extrinsic	O
apoptotic	O
pathways	O
.	O

The	O
other	O
induced	O
genes	O
revealed	O
a	O
whole	O
set	O
of	O
biologically	O
meaningful	O
genes	O
related	O
to	O
macrophage	B-Cell
activation	O
and	O
TH1	B-Gene_or_gene_product
immune	O
response	O
.	O

Immunohistochemical	O
analysis	O
suggested	O
a	O
specific	O
activation	O
or	O
differentiation	O
of	O
resident	O
macrophages	B-Cell
by	O
apoptotic	O
cells	B-Cell
.	O

These	O
biologic	O
insights	O
are	O
important	O
arguments	O
to	O
advocate	O
the	O
use	O
of	O
low	O
-	O
dose	O
radiotherapy	O
as	O
an	O
effective	O
palliative	O
treatment	O
for	O
follicular	B-Cancer
lymphoma	I-Cancer
.	O

Moreover	O
,	O
this	O
study	O
is	O
the	O
first	O
in	O
vivo	O
report	O
of	O
the	O
radiation	O
-	O
induced	O
p53	B-Gene_or_gene_product
apoptotic	O
response	O
in	O
patients	B-Organism
and	O
suggests	O
that	O
this	O
apoptotic	O
response	O
is	O
not	O
immunologically	O
silent	O
.	O

Caffeine	B-Simple_chemical
inhibits	O
adenosine	B-Simple_chemical
-	O
induced	O
accumulation	O
of	O
hypoxia	B-Gene_or_gene_product
-	I-Gene_or_gene_product
inducible	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
,	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
,	O
and	O
interleukin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
expression	O
in	O
hypoxic	O
human	B-Organism
colon	B-Cell
cancer	I-Cell
cells	I-Cell
.	O

Frequent	O
coffee	O
consumption	O
has	O
been	O
associated	O
with	O
a	O
reduced	O
risk	O
of	O
colorectal	B-Cancer
cancer	I-Cancer
in	O
a	O
number	O
of	O
case	O
-	O
control	O
studies	O
.	O

Coffee	O
is	O
a	O
leading	O
source	O
of	O
methylxanthines	B-Simple_chemical
,	O
such	O
as	O
caffeine	B-Simple_chemical
.	O

The	O
induction	O
of	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
and	O
interleukin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
(	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
)	O
is	O
an	O
essential	O
feature	O
of	O
tumor	B-Cancer
angiogenesis	O
,	O
and	O
the	O
hypoxia	B-Gene_or_gene_product
-	I-Gene_or_gene_product
inducible	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
transcription	O
factor	O
is	O
known	O
to	O
be	O
a	O
key	O
regulator	O
of	O
this	O
process	O
.	O

In	O
this	O
study	O
,	O
we	O
investigated	O
the	O
effects	O
of	O
caffeine	B-Simple_chemical
on	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
protein	O
accumulation	O
and	O
on	O
VEGF	B-Gene_or_gene_product
and	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
expression	O
in	O
the	O
human	B-Organism
colon	B-Cell
cancer	I-Cell
cell	I-Cell
line	I-Cell
HT29	I-Cell
under	O
hypoxic	O
conditions	O
.	O

Our	O
results	O
show	O
that	O
caffeine	B-Simple_chemical
significantly	O
inhibits	O
adenosine	B-Simple_chemical
-	O
induced	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
protein	O
accumulation	O
in	O
cancer	B-Cell
cells	I-Cell
.	O

We	O
show	O
that	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
and	O
VEGF	B-Gene_or_gene_product
are	O
increased	O
through	O
A3	B-Gene_or_gene_product
adenosine	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
stimulation	O
,	O
whereas	O
the	O
effects	O
on	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
are	O
mediated	O
via	O
the	O
A2B	B-Gene_or_gene_product
subtype	O
.	O

Pretreatment	O
of	O
cells	B-Cell
with	O
caffeine	B-Simple_chemical
significantly	O
reduces	O
adenosine	B-Simple_chemical
-	O
induced	O
VEGF	B-Gene_or_gene_product
promoter	O
activity	O
and	O
VEGF	B-Gene_or_gene_product
and	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
expression	O
.	O

The	O
mechanism	O
of	O
caffeine	B-Simple_chemical
seems	O
to	O
involve	O
the	O
inhibition	O
of	O
the	O
extracellular	B-Gene_or_gene_product
signal	I-Gene_or_gene_product
-	I-Gene_or_gene_product
regulated	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
1	I-Gene_or_gene_product
/	O
2	B-Gene_or_gene_product
(	O
ERK1	B-Gene_or_gene_product
/	O
2	B-Gene_or_gene_product
)	O
,	O
p38	B-Gene_or_gene_product
,	O
and	O
Akt	B-Gene_or_gene_product
,	O
leading	O
to	O
a	O
marked	O
decrease	O
in	O
adenosine	B-Simple_chemical
-	O
induced	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
accumulation	O
,	O
VEGF	B-Gene_or_gene_product
transcriptional	O
activation	O
,	O
and	O
VEGF	B-Gene_or_gene_product
and	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
protein	O
accumulation	O
.	O

From	O
a	O
functional	O
perspective	O
,	O
we	O
observe	O
that	O
caffeine	B-Simple_chemical
also	O
significantly	O
inhibits	O
the	O
A3	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
-	O
stimulated	O
cell	B-Cell
migration	O
of	O
colon	B-Cell
cancer	I-Cell
cells	I-Cell
.	O

Conditioned	O
media	O
prepared	O
from	O
colon	B-Cell
cells	I-Cell
treated	O
with	O
an	O
adenosine	B-Simple_chemical
analog	O
increased	O
human	B-Cell
umbilical	I-Cell
vein	I-Cell
endothelial	I-Cell
cell	I-Cell
migration	O
.	O

These	O
data	O
provide	O
evidence	O
that	O
adenosine	B-Simple_chemical
could	O
modulate	O
the	O
migration	O
of	O
colon	B-Cell
cancer	I-Cell
cells	I-Cell
by	O
an	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
/	O
VEGF	B-Gene_or_gene_product
/	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
-	O
dependent	O
mechanism	O
and	O
that	O
caffeine	B-Simple_chemical
has	O
the	O
potential	O
to	O
inhibit	O
colon	B-Cell
cancer	I-Cell
cell	I-Cell
growth	O
.	O

Medical	O
therapy	O
for	O
intermittent	O
claudication	O
.	O

Medical	O
therapy	O
to	O
improve	O
symptoms	O
,	O
stabilise	O
the	O
underlying	O
vascular	B-Multi-tissue_structure
disease	O
and	O
improve	O
lower	B-Organism_subdivision
limb	I-Organism_subdivision
outcomes	O
is	O
an	O
important	O
and	O
effective	O
adjunct	O
to	O
lifestyle	O
modification	O
and	O
surgical	O
or	O
endovascular	B-Immaterial_anatomical_entity
interventions	O
in	O
patients	B-Organism
with	O
IC	O
.	O

Randomised	O
placebo	O
controlled	O
trials	O
have	O
shown	O
that	O
the	O
phosphodiesterase	B-Gene_or_gene_product
III	I-Gene_or_gene_product
inhibitor	O
cilostazol	B-Simple_chemical
100mg	O
bid	O
improves	O
pain	O
-	O
free	O
and	O
maximum	O
walking	O
distance	O
,	O
as	O
well	O
as	O
quality	O
of	O
life	O
,	O
in	O
a	O
range	O
of	O
patients	B-Organism
with	O
intermittent	O
claudication	O
in	O
whom	O
there	O
is	O
no	O
evidence	O
of	O
tissue	B-Tissue
necrosis	O
or	O
rest	O
pain	O
.	O

This	O
review	O
summarises	O
the	O
evidence	O
from	O
8	O
pivotal	O
trials	O
of	O
cilostazol	B-Simple_chemical
involving	O
over	O
2000	O
patients	B-Organism
with	O
intermittent	O
claudication	O
treated	O
for	O
up	O
to	O
6	O
months	O
.	O

There	O
is	O
comparatively	O
less	O
evidence	O
to	O
support	O
the	O
use	O
of	O
other	O
treatment	O
modalities	O
for	O
relief	O
of	O
symptoms	O
in	O
intermittent	O
claudication	O
,	O
but	O
there	O
is	O
considerable	O
interest	O
in	O
therapeutic	O
angiogenesis	O
to	O
promote	O
new	O
vessel	B-Multi-tissue_structure
formation	O
and	O
enhance	O
collateralisation	O
of	O
the	O
lower	B-Organism_subdivision
limb	I-Organism_subdivision
using	O
recombinant	O
growth	O
factor	O
proteins	O
or	O
gene	O
transfer	O
strategies	O
.	O

The	O
rationale	O
for	O
therapeutic	O
angiogenesis	O
is	O
discussed	O
,	O
together	O
with	O
the	O
most	O
recent	O
results	O
from	O
randomised	O
trials	O
in	O
patients	B-Organism
with	O
peripheral	B-Multi-tissue_structure
arterial	I-Multi-tissue_structure
disease	O
.	O

Modulating	O
metastasis	O
by	O
a	O
lymphangiogenic	O
switch	O
in	O
prostate	B-Cancer
cancer	I-Cancer
.	O

Prostate	B-Cancer
cancer	I-Cancer
dissemination	O
is	O
difficult	O
to	O
detect	O
in	O
the	O
clinic	O
,	O
and	O
few	O
treatment	O
options	O
exist	O
for	O
patients	B-Organism
with	O
advanced	O
-	O
stage	O
disease	O
.	O

Our	O
aim	O
was	O
to	O
investigate	O
the	O
role	O
of	O
tumor	B-Cancer
lymphangiogenesis	O
during	O
metastasis	O
.	O

Further	O
,	O
we	O
implemented	O
a	O
noninvasive	O
molecular	O
imaging	O
technique	O
to	O
facilitate	O
the	O
assessment	O
of	O
the	O
metastatic	O
process	O
.	O

The	O
metastatic	O
potentials	O
of	O
several	O
human	B-Organism
prostate	B-Cancer
cancer	I-Cancer
xenograft	I-Cancer
models	O
,	O
LAPC	B-Cell
-	I-Cell
4	I-Cell
,	O
LAPC	B-Cell
-	I-Cell
9	I-Cell
,	O
PC3	B-Cell
and	O
CWR22Rv	B-Cell
-	I-Cell
1	I-Cell
were	O
compared	O
.	O

The	O
cells	B-Cell
were	O
labeled	O
with	O
luciferase	B-Gene_or_gene_product
,	O
a	O
bioluminescence	O
imaging	O
reporter	O
gene	O
,	O
to	O
enable	O
optical	O
imaging	O
.	O

After	O
tumor	B-Cancer
implantation	O
the	O
animals	B-Organism
were	O
examined	O
weekly	O
during	O
several	O
months	O
for	O
the	O
appearance	O
of	O
metastases	O
.	O

Metastatic	B-Cancer
lesions	I-Cancer
were	O
confirmed	O
by	O
immunohistochemistry	O
.	O

Additionally	O
,	O
the	O
angiogenic	O
and	O
lymphangiogenic	O
profiles	O
of	O
the	O
tumors	B-Cancer
were	O
characterized	O
.	O

To	O
confirm	O
the	O
role	O
of	O
lymphangiogenesis	O
in	O
mediating	O
metastasis	O
,	O
the	O
low	O
-	O
metastatic	O
LAPC	B-Cell
-	I-Cell
9	I-Cell
tumor	I-Cell
cells	I-Cell
were	O
engineered	O
to	O
overexpress	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
,	O
and	O
the	O
development	O
of	O
metastases	B-Cancer
was	O
evaluated	O
.	O

Our	O
results	O
show	O
CWR22Rv	B-Cell
-	I-Cell
1	I-Cell
and	O
PC3	B-Cell
tumor	I-Cell
cell	I-Cell
lines	I-Cell
to	O
be	O
more	O
metastatic	O
than	O
LAPC	B-Cell
-	I-Cell
4	I-Cell
,	O
which	O
in	O
turn	O
disseminates	O
more	O
readily	O
than	O
LAPC	B-Cell
-	I-Cell
9	I-Cell
.	O

The	O
difference	O
in	O
metastatic	O
potential	O
correlated	O
with	O
the	O
endogenous	O
production	O
levels	O
of	O
lymphangiogenic	O
growth	O
factor	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
and	O
the	O
presence	O
of	O
tumor	B-Multi-tissue_structure
lymphatics	I-Multi-tissue_structure
.	O

In	O
agreement	O
,	O
induced	O
overexpression	O
of	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
in	O
LAPC	B-Cell
-	I-Cell
9	I-Cell
enhanced	O
tumor	B-Cancer
lymphangiogenesis	O
leading	O
to	O
the	O
development	O
of	O
metastatic	B-Cancer
lesions	I-Cancer
.	O

Taken	O
together	O
,	O
our	O
studies	O
,	O
based	O
on	O
a	O
molecular	O
imaging	O
approach	O
for	O
semiquantitative	O
detection	O
of	O
micrometastases	O
,	O
point	O
to	O
an	O
important	O
role	O
of	O
tumor	B-Multi-tissue_structure
lymphatics	I-Multi-tissue_structure
in	O
the	O
metastatic	O
process	O
of	O
human	B-Organism
prostate	B-Cancer
cancer	I-Cancer
.	O

In	O
particular	O
,	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
seems	O
to	O
play	O
a	O
key	O
role	O
in	O
prostate	B-Cancer
cancer	I-Cancer
metastasis	O
.	O

An	O
assay	O
to	O
measure	O
angiogenesis	O
in	O
human	B-Organism
fat	B-Tissue
tissue	I-Tissue
.	O

BACKGROUND	O
:	O
Inhibition	O
of	O
angiogenesis	O
reverses	O
rodent	O
obesity	O
.	O

A	O
validated	O
assay	O
in	O
human	B-Organism
fat	B-Tissue
tissue	I-Tissue
is	O
needed	O
to	O
study	O
the	O
role	O
of	O
angiogenesis	O
in	O
human	B-Organism
obesity	O
.	O

METHODS	O
:	O
Human	B-Organism
fat	B-Tissue
tissue	I-Tissue
fragments	O
from	O
surgery	O
were	O
placed	O
in	O
96	O
-	O
well	O
plates	O
,	O
embedded	O
in	O
fibrin	B-Gene_or_gene_product
thrombin	B-Gene_or_gene_product
clot	B-Organism_substance
and	O
overlaid	O
with	O
cell	B-Cell
culture	O
media	O
containing	O
20	O
%	O
fetal	O
bovine	B-Organism
serum	B-Organism_substance
.	O

After	O
15	O
days	O
,	O
the	O
clots	B-Organism_substance
were	O
examined	O
by	O
histology	O
and	O
electron	O
microscopy	O
.	O

The	O
effect	O
of	O
taxol	B-Simple_chemical
,	O
cobalt	B-Simple_chemical
chloride	I-Simple_chemical
and	O
a	O
heparin	B-Simple_chemical
-	I-Simple_chemical
steroid	I-Simple_chemical
combination	O
was	O
tested	O
in	O
the	O
fat	B-Tissue
tissue	I-Tissue
assay	O
and	O
compared	O
to	O
the	O
validated	O
human	B-Organism
placental	B-Multi-tissue_structure
vein	I-Multi-tissue_structure
angiogenesis	O
model	O
(	O
HPVAM	O
)	O
.	O

RESULTS	O
:	O
Blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
initiated	O
growth	O
and	O
elongated	O
from	O
the	O
fat	B-Tissue
tissue	I-Tissue
fragments	O
over	O
15	O
days	O
.	O

Presence	O
of	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
was	O
confirmed	O
with	O
histology	O
and	O
electron	O
microscopy	O
.	O

Taxol	B-Simple_chemical
at	O
10	O
(	O
-	O
6	O
)	O
and	O
10	O
(	O
-	O
7	O
)	O
M	O
completely	O
inhibited	O
angiogenesis	O
,	O
while	O
Taxol	B-Simple_chemical
10	O
(	O
-	O
8	O
)	O
and	O
10	O
(	O
-	O
9	O
)	O
M	O
and	O
the	O
heparin	B-Simple_chemical
-	I-Simple_chemical
steroid	I-Simple_chemical
partially	O
inhibited	O
angiogenesis	O
.	O

The	O
response	O
to	O
taxol	B-Simple_chemical
and	O
heparin	B-Simple_chemical
-	I-Simple_chemical
steroid	I-Simple_chemical
was	O
similar	O
to	O
that	O
of	O
the	O
HPVAM	O
,	O
a	O
validated	O
angiogenesis	O
assay	O
.	O

Cobalt	B-Simple_chemical
chloride	I-Simple_chemical
,	O
a	O
stimulator	O
of	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
stimulated	O
angiogenesis	O
initiation	O
at	O
10	O
(	O
-	O
9	O
)	O
M	O
in	O
fat	B-Tissue
tissue	I-Tissue
and	O
the	O
HPVAM	O
,	O
but	O
at	O
10	O
(	O
-	O
10	O
)	O
M	O
blood	B-Multi-tissue_structure
vessel	I-Multi-tissue_structure
growth	O
was	O
stimulated	O
only	O
in	O
the	O
fat	B-Tissue
assay	O
.	O

CONCLUSION	O
:	O
This	O
angiogenesis	O
assay	O
based	O
on	O
human	B-Organism
fat	B-Tissue
tissue	I-Tissue
uses	O
three	O
-	O
dimensionally	O
intact	O
human	B-Organism
tissue	B-Tissue
.	O

The	O
vessels	B-Multi-tissue_structure
are	O
derived	O
from	O
quiescient	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
within	O
the	O
fat	B-Tissue
.	O

These	O
properties	O
allow	O
the	O
angiogenic	O
switch	O
to	O
be	O
evaluated	O
in	O
an	O
in	O
vitro	O
setting	O
.	O

The	O
angiogenic	O
response	O
of	O
fat	B-Tissue
tissue	I-Tissue
is	O
not	O
identical	O
to	O
placental	B-Tissue
tissue	I-Tissue
.	O

This	O
assay	O
allows	O
exploration	O
of	O
angiogenesis	O
in	O
fat	B-Tissue
tissue	I-Tissue
.	O

Subcellular	O
localisation	O
of	O
BAG	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
its	O
regulation	O
of	O
vitamin	B-Gene_or_gene_product
D	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
-	O
mediated	O
transactivation	O
and	O
involucrin	B-Gene_or_gene_product
expression	O
in	O
oral	B-Cell
keratinocytes	I-Cell
:	O
implications	O
for	O
oral	B-Organism_subdivision
carcinogenesis	O
.	O

In	O
oral	B-Cancer
cancers	I-Cancer
,	O
cytoplasmic	B-Organism_substance
BAG	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
overexpression	O
is	O
a	O
marker	O
of	O
poor	O
prognosis	O
.	O

BAG	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
regulates	O
cellular	B-Cell
growth	O
,	O
differentiation	O
and	O
survival	O
through	O
interactions	O
with	O
diverse	O
proteins	O
,	O
including	O
the	O
vitamin	B-Gene_or_gene_product
D	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
(	O
VDR	B-Gene_or_gene_product
)	O
,	O
a	O
key	O
regulator	O
of	O
keratinocyte	B-Cell
growth	O
and	O
differentiation	O
.	O

BAG	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
is	O
expressed	O
ubiquitously	O
in	O
human	B-Organism
cells	B-Cell
as	O
three	O
major	O
isoforms	O
of	O
50	O
kDa	O
(	O
BAG	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1L	I-Gene_or_gene_product
)	O
,	O
46	O
kDa	O
(	O
BAG	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1M	I-Gene_or_gene_product
)	O
and	O
36	O
kDa	O
(	O
BAG	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1S	I-Gene_or_gene_product
)	O
from	O
a	O
single	O
mRNA	O
.	O

In	O
oral	B-Cell
keratinocytes	I-Cell
BAG	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1L	I-Gene_or_gene_product
,	O
but	O
not	O
BAG	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1M	I-Gene_or_gene_product
and	O
BAG	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1S	I-Gene_or_gene_product
,	O
enhanced	O
VDR	B-Gene_or_gene_product
transactivation	O
in	O
response	O
to	O
1alpha	B-Simple_chemical
,	I-Simple_chemical
25	I-Simple_chemical
-	I-Simple_chemical
dihydroxyvitamin	I-Simple_chemical
D3	I-Simple_chemical
.	O

BAG	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1L	I-Gene_or_gene_product
was	O
nucleoplasmic	B-Organism_substance
and	O
nucleolar	B-Cellular_component
,	O
whereas	O
BAG	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1S	I-Gene_or_gene_product
and	O
BAG	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1M	I-Gene_or_gene_product
were	O
cytoplasmic	B-Organism_substance
and	O
nucleoplasmic	B-Organism_substance
in	O
localisation	O
.	O

Having	O
identified	O
the	O
nucleolar	B-Cellular_component
localisation	O
sequence	O
in	O
BAG	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1L	I-Gene_or_gene_product
,	O
we	O
showed	O
that	O
mutation	O
of	O
this	O
sequence	O
did	O
not	O
prevent	O
BAG	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1L	I-Gene_or_gene_product
from	O
potentiating	O
VDR	B-Gene_or_gene_product
activity	O
.	O

BAG	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1L	I-Gene_or_gene_product
also	O
potentiated	O
transactivation	O
of	O
known	O
vitamin	B-Simple_chemical
-	I-Simple_chemical
D	I-Simple_chemical
-	O
responsive	O
gene	O
promoters	O
,	O
osteocalcin	B-Gene_or_gene_product
and	O
24	B-Gene_or_gene_product
-	I-Gene_or_gene_product
hydroxylase	I-Gene_or_gene_product
,	O
and	O
enhanced	O
VDR	B-Gene_or_gene_product
-	O
dependent	O
transcription	O
and	O
protein	O
expression	O
of	O
the	O
keratinocyte	B-Cell
differentiation	O
marker	O
,	O
involucrin	B-Gene_or_gene_product
.	O

These	O
results	O
demonstrate	O
endogenous	O
gene	O
regulation	O
by	O
BAG	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1L	I-Gene_or_gene_product
by	O
potentiating	O
nuclear	B-Gene_or_gene_product
hormone	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
function	O
and	O
suggest	O
a	O
role	O
for	O
BAG	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1L	I-Gene_or_gene_product
in	O
24	B-Gene_or_gene_product
-	I-Gene_or_gene_product
hydroxylase	I-Gene_or_gene_product
regulation	O
of	O
vitamin	B-Simple_chemical
D	I-Simple_chemical
metabolism	O
and	O
the	O
cellular	B-Cell
response	O
of	O
oral	B-Cell
keratinocytes	I-Cell
to	O
1alpha	B-Simple_chemical
,	I-Simple_chemical
25	I-Simple_chemical
-	I-Simple_chemical
dihydroxyvitamin	I-Simple_chemical
D3	I-Simple_chemical
.	O

By	O
contrast	O
to	O
the	O
cytoplasmic	B-Organism_substance
BAG	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
isoforms	O
,	O
BAG	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1L	I-Gene_or_gene_product
may	O
act	O
to	O
suppress	O
tumorigenesis	O
.	O

Transporters	O
,	O
enzymes	O
,	O
and	O
enalapril	B-Simple_chemical
removal	O
in	O
a	O
rat	B-Organism
(	O
CC531	B-Cell
-	O
induced	O
)	O
liver	B-Organ
metastatic	O
model	O
.	O

Temporal	O
changes	O
in	O
physiological	O
spaces	O
,	O
protein	O
expression	O
of	O
transporters	O
and	O
enzymes	O
,	O
and	O
enalapril	B-Simple_chemical
removal	O
were	O
appraised	O
in	O
the	O
metastatic	O
liver	B-Cancer
tumor	I-Cancer
model	O
developed	O
from	O
male	O
Wag	B-Organism
/	I-Organism
Rij	I-Organism
rats	I-Organism
after	O
the	O
intraportal	O
injection	O
of	O
CC531	B-Cell
colon	I-Cell
adenocarcinoma	I-Cell
cells	I-Cell
;	O
sham	O
-	O
operated	O
preparations	O
received	O
PBS	O
.	O

Liver	B-Tissue
tissue	I-Tissue
spaces	O
,	O
investigated	O
with	O
multiple	O
indicator	O
dilution	O
technique	O
in	O
liver	B-Organ
perfusion	O
studies	O
,	O
were	O
unchanged	O
at	O
week	O
3	O
after	O
tumor	B-Cancer
induction	O
.	O

At	O
week	O
4	O
,	O
however	O
,	O
the	O
sinusoidal	B-Multi-tissue_structure
blood	I-Multi-tissue_structure
volume	O
and	O
albumin	B-Gene_or_gene_product
Disse	O
space	O
in	O
tumor	B-Cancer
-	O
bearing	O
livers	B-Organ
were	O
slightly	O
lower	O
compared	O
with	O
those	O
of	O
shams	O
.	O

Increased	O
levels	O
of	O
the	O
canalicular	B-Multi-tissue_structure
ATP	B-Simple_chemical
transporters	O
,	O
P	O
-	O
glycoprotein	O
,	O
multidrug	B-Gene_or_gene_product
resistance	I-Gene_or_gene_product
-	I-Gene_or_gene_product
associated	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
2	I-Gene_or_gene_product
(	O
Mrp2	B-Gene_or_gene_product
)	O
,	O
and	O
bile	B-Simple_chemical
salt	I-Simple_chemical
export	O
pump	O
(	O
Bsep	O
)	O
at	O
week	O
2	O
(	O
P	O
<	O
0	O
.	O
05	O
)	O
,	O
unchanged	O
levels	O
of	O
Ntcp	B-Gene_or_gene_product
,	O
Oatp1a1	B-Gene_or_gene_product
,	O
Oatp1a4	B-Gene_or_gene_product
,	O
and	O
Mct2	B-Gene_or_gene_product
,	O
but	O
decreased	O
levels	O
of	O
cytochrome	B-Gene_or_gene_product
P450	I-Gene_or_gene_product
3a2	I-Gene_or_gene_product
(	O
Cyp3a2	B-Gene_or_gene_product
)	O
and	O
glutathione	B-Gene_or_gene_product
S	I-Gene_or_gene_product
-	I-Gene_or_gene_product
transferase	I-Gene_or_gene_product
(	O
Gst4	B-Gene_or_gene_product
-	I-Gene_or_gene_product
4	I-Gene_or_gene_product
)	O
at	O
week	O
4	O
(	O
P	O
<	O
0	O
.	O
05	O
)	O
were	O
observed	O
in	O
peritumor	B-Cell
vs	O
.	O
sham	O
-	O
operated	O
liver	B-Tissue
tissues	I-Tissue
with	O
Western	O
blotting	O
.	O

The	O
steady	O
-	O
state	O
extraction	O
ratio	O
of	O
enalapril	B-Simple_chemical
,	O
a	O
substrate	O
that	O
enters	O
the	O
liver	B-Organ
rapidly	O
via	O
Oatp1a1	B-Gene_or_gene_product
and	O
primarily	O
undergoes	O
metabolism	O
by	O
the	O
carboxylesterases	O
,	O
was	O
unaffected	O
by	O
liver	B-Organ
metastasis	O
at	O
week	O
4	O
regardless	O
of	O
its	O
delivery	O
via	O
the	O
portal	B-Multi-tissue_structure
vein	I-Multi-tissue_structure
or	O
hepatic	B-Multi-tissue_structure
artery	I-Multi-tissue_structure
into	O
the	O
perfused	O
liver	B-Organ
preparations	O
.	O

Wnt	B-Gene_or_gene_product
signaling	O
,	O
stem	B-Cell
cells	I-Cell
,	O
and	O
the	O
cellular	B-Cell
origin	O
of	O
breast	B-Cancer
cancer	I-Cancer
.	O

The	O
breast	B-Tissue
epithelium	I-Tissue
comprises	O
cells	B-Cell
at	O
different	O
stages	O
of	O
differentiation	O
,	O
including	O
stem	B-Cell
cells	I-Cell
,	O
progenitor	B-Cell
cells	I-Cell
,	O
and	O
more	O
differentiated	O
epithelial	B-Cell
and	O
myoepithelial	B-Cell
cells	I-Cell
.	O

Wnt	B-Gene_or_gene_product
signaling	O
plays	O
a	O
critical	O
role	O
in	O
regulating	O
stem	B-Cell
/	O
progenitor	B-Cell
cells	I-Cell
in	O
the	O
mammary	B-Organ
gland	I-Organ
as	O
well	O
as	O
other	O
tissue	B-Multi-tissue_structure
compartments	I-Multi-tissue_structure
.	O

Furthermore	O
,	O
there	O
is	O
strong	O
evidence	O
suggesting	O
that	O
aberrant	O
activation	O
of	O
Wnt	B-Gene_or_gene_product
signaling	O
induces	O
mammary	B-Cancer
tumors	I-Cancer
from	O
stem	B-Cell
/	O
progenitor	B-Cell
cells	I-Cell
,	O
and	O
that	O
Wnt	B-Gene_or_gene_product
exerts	O
its	O
oncogenic	O
effects	O
through	O
LRP5	B-Gene_or_gene_product
/	O
6	B-Gene_or_gene_product
-	O
mediated	O
activation	O
of	O
beta	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
and	O
mTOR	B-Gene_or_gene_product
pathways	O
.	O

Recent	O
studies	O
using	O
avian	B-Organism
retrovirus	I-Organism
-	O
mediated	O
introduction	O
of	O
oncogenes	O
into	O
a	O
small	O
subset	O
of	O
somatic	B-Cell
mammary	I-Cell
cells	I-Cell
suggest	O
that	O
polyoma	B-Organism
middle	B-Gene_or_gene_product
T	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
(	O
PyMT	B-Gene_or_gene_product
)	O
may	O
also	O
preferentially	O
transform	O
stem	B-Cell
/	O
progenitor	B-Cell
cells	I-Cell
.	O

These	O
observations	O
suggest	O
that	O
stem	B-Cell
/	O
progenitor	B-Cell
cells	I-Cell
in	O
the	O
mammary	B-Organ
gland	I-Organ
may	O
be	O
especially	O
susceptible	O
to	O
oncogenic	O
transformation	O
.	O

Whether	O
more	O
differentiated	O
cells	B-Cell
may	O
also	O
be	O
transformed	O
by	O
particular	O
oncogenes	O
is	O
actively	O
debated	O
;	O
it	O
is	O
presently	O
unclear	O
whether	O
stem	B-Cell
cells	I-Cell
or	O
differentiated	O
mammary	B-Cell
cells	I-Cell
are	O
more	O
susceptible	O
to	O
transformation	O
by	O
individual	O
oncogenes	O
.	O

Better	O
stem	B-Cell
cell	I-Cell
and	O
progenitor	B-Cell
cell	I-Cell
markers	O
as	O
well	O
as	O
the	O
ability	O
to	O
specifically	O
target	O
oncogenes	O
into	O
different	O
mammary	B-Cell
cell	I-Cell
types	O
will	O
be	O
needed	O
to	O
determine	O
the	O
spectrum	O
of	O
oncogene	O
transformation	O
for	O
stem	B-Cell
cells	I-Cell
versus	O
more	O
differentiated	O
cells	B-Cell
.	O

Ovarian	B-Cancer
cancers	I-Cancer
overexpress	O
the	O
antimicrobial	O
protein	O
hCAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
18	I-Gene_or_gene_product
and	O
its	O
derivative	O
LL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
37	I-Gene_or_gene_product
increases	O
ovarian	B-Cell
cancer	I-Cell
cell	I-Cell
proliferation	O
and	O
invasion	O
.	O

The	O
role	O
of	O
the	O
pro	O
-	O
inflammatory	O
peptide	O
,	O
LL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
37	I-Gene_or_gene_product
,	O
and	O
its	O
pro	O
-	O
form	O
,	O
human	B-Gene_or_gene_product
cationic	I-Gene_or_gene_product
antimicrobial	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
18	I-Gene_or_gene_product
(	O
hCAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
18	I-Gene_or_gene_product
)	O
,	O
in	O
cancer	B-Cancer
development	O
and	O
progression	O
is	O
poorly	O
understood	O
.	O

In	O
damaged	O
and	O
inflamed	O
tissue	B-Tissue
,	O
LL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
37	I-Gene_or_gene_product
functions	O
as	O
a	O
chemoattractant	O
,	O
mitogen	O
and	O
pro	O
-	O
angiogenic	O
factor	O
suggesting	O
that	O
the	O
peptide	O
may	O
potentiate	O
tumor	B-Cancer
progression	O
.	O

The	O
aim	O
of	O
this	O
study	O
was	O
to	O
characterize	O
the	O
distribution	O
of	O
hCAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
18	I-Gene_or_gene_product
/	O
LL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
37	I-Gene_or_gene_product
in	O
normal	O
and	O
cancerous	O
ovarian	B-Tissue
tissue	I-Tissue
and	O
to	O
examine	O
the	O
effects	O
of	O
LL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
37	I-Gene_or_gene_product
on	O
ovarian	B-Cell
cancer	I-Cell
cells	I-Cell
.	O

Expression	O
of	O
hCAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
18	I-Gene_or_gene_product
/	O
LL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
37	I-Gene_or_gene_product
was	O
localized	O
to	O
immune	B-Cell
and	O
granulosa	B-Cell
cells	I-Cell
of	O
normal	B-Tissue
ovarian	I-Tissue
tissue	I-Tissue
.	O

By	O
contrast	O
,	O
ovarian	B-Cancer
tumors	I-Cancer
displayed	O
significantly	O
higher	O
levels	O
of	O
hCAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
18	I-Gene_or_gene_product
/	O
LL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
37	I-Gene_or_gene_product
where	O
expression	O
was	O
observed	O
in	O
tumor	B-Cell
and	O
stromal	B-Cell
cells	I-Cell
.	O

Protein	O
expression	O
was	O
statistically	O
compared	O
to	O
the	O
degree	O
of	O
immune	B-Cell
cell	I-Cell
infiltration	O
and	O
microvessel	B-Tissue
density	O
in	O
epithelial	B-Cancer
-	I-Cancer
derived	I-Cancer
ovarian	I-Cancer
tumors	I-Cancer
and	O
a	O
significant	O
correlation	O
was	O
observed	O
for	O
both	O
.	O

It	O
was	O
demonstrated	O
that	O
ovarian	B-Organism_substance
tumor	I-Organism_substance
tissue	I-Organism_substance
lysates	I-Organism_substance
and	O
ovarian	B-Cell
cancer	I-Cell
cell	I-Cell
lines	I-Cell
express	O
hCAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
18	I-Gene_or_gene_product
/	O
LL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
37	I-Gene_or_gene_product
.	O

Treatment	O
of	O
ovarian	B-Cell
cancer	I-Cell
cell	I-Cell
lines	I-Cell
with	O
recombinant	O
LL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
37	I-Gene_or_gene_product
stimulated	O
proliferation	O
,	O
chemotaxis	O
,	O
invasion	O
and	O
matrix	B-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
expression	O
.	O

These	O
data	O
demonstrate	O
for	O
the	O
first	O
time	O
that	O
hCAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
18	I-Gene_or_gene_product
/	O
LL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
37	I-Gene_or_gene_product
is	O
significantly	O
overexpressed	O
in	O
ovarian	B-Cancer
tumors	I-Cancer
and	O
suggest	O
LL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
37	I-Gene_or_gene_product
may	O
contribute	O
to	O
ovarian	B-Organ
tumorigenesis	O
through	O
direct	O
stimulation	O
of	O
tumor	B-Cell
cells	I-Cell
,	O
initiation	O
of	O
angiogenesis	O
and	O
recruitment	O
of	O
immune	B-Cell
cells	I-Cell
.	O

These	O
data	O
provide	O
further	O
evidence	O
of	O
the	O
existing	O
relationship	O
between	O
pro	O
-	O
inflammatory	O
molecules	O
and	O
ovarian	B-Cancer
cancer	I-Cancer
progression	O
.	O

Tumor	B-Cancer
microenvironment	O
,	O
a	O
dangerous	O
society	O
leading	O
to	O
cancer	B-Cancer
metastasis	O
.	O

From	O
mechanisms	O
to	O
therapy	O
and	O
prevention	O
.	O

Cancer	B-Cancer
is	O
no	O
longer	O
considered	O
by	O
scientists	O
just	O
a	O
jumble	O
of	O
mutated	O
cells	B-Cell
.	O

To	O
grow	O
,	O
invade	O
and	O
metastasize	O
,	O
a	O
treacherous	O
society	O
between	O
cancer	B-Cancer
and	O
host	B-Cell
cells	I-Cell
must	O
be	O
formed	O
,	O
and	O
this	O
association	O
provides	O
novel	O
and	O
effective	O
clinical	O
targets	O
for	O
cancer	B-Cancer
control	O
and	O
prevention	O
.	O

This	O
collection	O
of	O
reviews	O
at	O
the	O
front	O
-	O
edge	O
of	O
scientific	O
knowledge	O
focuses	O
on	O
host	B-Cell
-	O
tumor	B-Cell
cell	I-Cell
interactions	O
,	O
the	O
disastrous	O
consequences	O
they	O
can	O
produce	O
and	O
approaches	O
the	O
ways	O
to	O
break	O
up	O
these	O
cellular	B-Cell
conspiracies	O
,	O
to	O
leave	O
the	O
tumor	B-Cell
cells	I-Cell
unattended	O
and	O
vulnerable	O
.	O

[	O
Photodynamic	O
therapy	O
in	O
severe	O
chronic	O
central	O
serous	B-Organism_substance
chorioretinopaty	O
]	O

OBJECTIVE	O
:	O
To	O
determine	O
the	O
efficacy	O
of	O
Photodynamic	O
Therapy	O
(	O
PDT	O
)	O
in	O
chronic	O
Central	O
Serous	B-Organism_substance
Chorioretinopathy	O
(	O
CSC	O
)	O
.	O

METHODS	O
:	O
Patients	B-Organism
diagnosed	O
with	O
chronic	O
CSC	O
,	O
with	O
clinical	O
evidence	O
of	O
activity	O
and	O
treated	O
with	O
Photodynamic	O
Therapy	O
,	O
are	O
included	O
in	O
this	O
report	O
.	O

All	O
were	O
assessed	O
by	O
a	O
complete	O
ophthalmological	O
examination	O
,	O
including	O
assessment	O
of	O
the	O
best	O
corrected	O
visual	O
acuity	O
(	O
BCVA	O
)	O
using	O
an	O
ETDRS	O
chart	O
,	O
fluorescein	B-Simple_chemical
and	O
indocyanine	B-Simple_chemical
angiography	O
and	O
optical	O
coherence	O
tomography	O
(	O
OCT	O
)	O
.	O

The	O
main	O
objective	O
of	O
the	O
study	O
was	O
to	O
determine	O
the	O
mean	O
visual	O
acuity	O
change	O
.	O

RESULTS	O
:	O
11	O
eyes	B-Organ
of	O
11	O
patients	B-Organism
were	O
included	O
in	O
the	O
study	O
,	O
which	O
had	O
a	O
mean	O
follow	O
-	O
up	O
period	O
of	O
11	O
months	O
.	O

The	O
mean	O
BCVA	O
increased	O
from	O
20	O
/	O
76	O
to	O
20	O
/	O
64	O
.	O

35	O
%	O
of	O
eyes	B-Organ
improved	O
their	O
BCVA	O
by	O
2	O
lines	O
or	O
more	O
,	O
45	O
%	O
remained	O
stable	O
and	O
18	O
%	O
lost	O
2	O
lines	O
or	O
more	O
.	O

Choroidal	B-Multi-tissue_structure
hyperpermeability	O
was	O
reduced	O
in	O
every	O
case	O
.	O

Neurosensorial	O
retinal	B-Multi-tissue_structure
detachment	O
decreased	O
in	O
80	O
%	O
of	O
cases	O
.	O

Only	O
one	O
eye	B-Organ
received	O
a	O
second	O
PDT	O
treatment	O
due	O
to	O
choroidal	B-Multi-tissue_structure
neovascularization	O
.	O

An	O
increase	O
of	O
atrophy	O
over	O
the	O
Retinal	B-Tissue
Pigment	I-Tissue
Epithelium	I-Tissue
(	O
RPE	B-Tissue
)	O
was	O
observed	O
in	O
another	O
patient	B-Organism
.	O

CONCLUSIONS	O
:	O
PDT	O
can	O
reduce	O
the	O
clinical	O
signs	O
of	O
activity	O
,	O
such	O
as	O
choroidal	B-Multi-tissue_structure
hyperpermeability	O
or	O
neurosensorial	O
retinal	B-Multi-tissue_structure
detachment	O
,	O
in	O
patients	B-Organism
affected	O
by	O
chronic	O
CSC	O
.	O

However	O
,	O
the	O
increase	O
in	O
visual	O
acuity	O
is	O
variable	O
,	O
probably	O
due	O
to	O
the	O
extent	O
of	O
RPE	B-Tissue
damage	O
.	O

Angiopoietin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
prevents	O
VEGF	B-Gene_or_gene_product
-	O
induced	O
endothelial	B-Tissue
permeability	O
by	O
sequestering	O
Src	B-Gene_or_gene_product
through	O
mDia	B-Gene_or_gene_product
.	O

Vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
and	O
Angiopoietin	B-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
Ang1	B-Gene_or_gene_product
)	O
are	O
both	O
potent	O
proangiogenic	O
factors	O
,	O
but	O
,	O
whereas	O
VEGF	B-Gene_or_gene_product
causes	O
vascular	B-Multi-tissue_structure
permeability	O
,	O
Ang1	B-Gene_or_gene_product
stabilizes	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
and	O
protects	O
them	O
from	O
VEGF	B-Gene_or_gene_product
-	O
induced	O
plasma	B-Organism_substance
leakage	O
.	O

The	O
antivascular	B-Multi-tissue_structure
permeability	O
mechanisms	O
deployed	O
by	O
Ang1	B-Gene_or_gene_product
are	O
still	O
undefined	O
.	O

Here	O
,	O
we	O
demonstrate	O
that	O
Ang1	B-Gene_or_gene_product
halts	O
the	O
ability	O
of	O
VEGF	B-Gene_or_gene_product
to	O
induce	O
the	O
phosphorylation	O
-	O
dependent	O
redistribution	O
of	O
the	O
adhesion	O
molecule	O
VE	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
,	O
thereby	O
rescuing	O
the	O
endothelial	B-Tissue
barrier	I-Tissue
function	O
.	O

Ang1	B-Gene_or_gene_product
inhibits	O
the	O
activation	O
of	O
Src	B-Gene_or_gene_product
by	O
VEGF	B-Gene_or_gene_product
,	O
the	O
most	O
upstream	O
component	O
of	O
the	O
pathway	O
linking	O
VEGF	B-Gene_or_gene_product
receptors	O
to	O
VE	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
internalization	O
.	O

Indeed	O
,	O
Ang1	B-Gene_or_gene_product
promotes	O
the	O
activation	O
of	O
mDia	B-Gene_or_gene_product
through	O
RhoA	B-Gene_or_gene_product
,	O
resulting	O
in	O
the	O
association	O
of	O
mDia	B-Gene_or_gene_product
with	O
Src	B-Gene_or_gene_product
.	O

This	O
ultimately	O
deprives	O
VEGF	B-Gene_or_gene_product
receptors	O
of	O
an	O
essential	O
molecule	O
required	O
for	O
promoting	O
the	O
disruption	O
of	O
endothelial	B-Cell
cell	O
-	O
cell	O
contacts	O
and	O
paracellular	B-Immaterial_anatomical_entity
permeability	O
.	O

RUNX1	B-Gene_or_gene_product
DNA	B-Cellular_component
-	O
binding	O
mutations	O
and	O
RUNX1	B-Gene_or_gene_product
-	O
PRDM16	B-Gene_or_gene_product
cryptic	O
fusions	O
in	O
BCR	B-Gene_or_gene_product
-	O
ABL	B-Gene_or_gene_product
+	O
leukemias	O
are	O
frequently	O
associated	O
with	O
secondary	O
trisomy	O
21	B-Cellular_component
and	O
may	O
contribute	O
to	O
clonal	B-Cell
evolution	O
and	O
imatinib	B-Simple_chemical
resistance	O
.	O

Acquired	O
molecular	O
abnormalities	O
(	O
mutations	O
or	O
chromosomal	B-Cellular_component
translocations	O
)	O
of	O
the	O
RUNX1	B-Gene_or_gene_product
transcription	O
factor	O
gene	O
are	O
frequent	O
in	O
acute	B-Cancer
myeloblastic	I-Cancer
leukemias	I-Cancer
(	O
AMLs	B-Cancer
)	O
and	O
in	O
therapy	O
-	O
related	O
myelodysplastic	O
syndromes	O
,	O
but	O
rarely	O
in	O
acute	B-Cancer
lymphoblastic	I-Cancer
leukemias	I-Cancer
(	O
ALLs	B-Cancer
)	O
and	O
chronic	B-Cancer
myelogenous	I-Cancer
leukemias	I-Cancer
(	O
CMLs	B-Cancer
)	O
.	O

Among	O
18	O
BCR	B-Gene_or_gene_product
-	O
ABL	B-Gene_or_gene_product
+	O
leukemias	O
presenting	O
acquired	O
trisomy	O
of	O
chromosome	B-Cellular_component
21	I-Cellular_component
,	O
we	O
report	O
a	O
high	O
frequency	O
(	O
33	O
%	O
)	O
of	O
recurrent	O
point	O
mutations	O
(	O
4	O
in	O
myeloid	B-Cancer
blast	I-Cancer
crisis	I-Cancer
[	I-Cancer
BC	I-Cancer
]	I-Cancer
CML	I-Cancer
and	O
one	O
in	O
chronic	B-Cancer
phase	I-Cancer
CML	I-Cancer
)	O
within	O
the	O
DNA	B-Cellular_component
-	O
binding	O
region	O
of	O
RUNX1	B-Gene_or_gene_product
.	O

We	O
did	O
not	O
found	O
any	O
mutation	O
in	O
de	O
novo	O
BCR	B-Gene_or_gene_product
-	O
ABL	B-Gene_or_gene_product
+	O
ALLs	O
or	O
lymphoid	B-Cancer
BC	I-Cancer
CML	I-Cancer
.	O

Emergence	O
of	O
the	O
RUNX1	B-Gene_or_gene_product
mutations	O
was	O
detected	O
at	O
diagnosis	O
or	O
before	O
the	O
acquisition	O
of	O
trisomy	O
21	B-Cellular_component
during	O
disease	O
progression	O
.	O

In	O
addition	O
,	O
we	O
also	O
report	O
a	O
high	O
frequency	O
of	O
cryptic	O
chromosomal	B-Cellular_component
RUNX1	B-Gene_or_gene_product
translocation	O
to	O
a	O
novel	O
recently	O
described	O
gene	O
partner	O
,	O
PRDM16	B-Gene_or_gene_product
on	O
chromosome	B-Cellular_component
1p36	I-Cellular_component
,	O
for	O
3	O
(	O
21	O
.	O
4	O
%	O
)	O
of	O
14	O
investigated	O
patients	B-Organism
:	O
2	O
myeloid	B-Cancer
BC	I-Cancer
CMLs	I-Cancer
and	O
,	O
for	O
the	O
first	O
time	O
,	O
1	O
therapy	O
-	O
related	O
BCR	B-Gene_or_gene_product
-	O
ABL	B-Gene_or_gene_product
+	O
ALL	O
.	O

Two	O
patients	B-Organism
presented	O
both	O
RUNX1	B-Gene_or_gene_product
mutations	O
and	O
RUNX1	B-Gene_or_gene_product
-	O
PRDM16	B-Gene_or_gene_product
fusion	O
.	O

These	O
events	O
are	O
associated	O
with	O
a	O
short	O
survival	O
and	O
support	O
the	O
concept	O
of	O
a	O
cooperative	O
effect	O
of	O
BCR	B-Gene_or_gene_product
-	O
ABL	B-Gene_or_gene_product
with	O
molecular	O
RUNX1	B-Gene_or_gene_product
abnormalities	O
on	O
the	O
differentiation	O
arrest	O
phenotype	O
observed	O
during	O
progression	O
of	O
CML	B-Cancer
and	O
in	O
BCR	B-Gene_or_gene_product
-	O
ABL	B-Gene_or_gene_product
+	O
ALL	O
.	O

Suppression	O
of	O
lung	B-Cancer
tumor	I-Cancer
growth	O
and	O
metastasis	O
in	O
mice	B-Organism
by	O
adeno	B-Organism
-	I-Organism
associated	I-Organism
virus	I-Organism
-	O
mediated	O
expression	O
of	O
vasostatin	B-Gene_or_gene_product
.	O

PURPOSE	O
:	O
Angiogenesis	O
inhibitors	O
have	O
strong	O
therapeutic	O
potential	O
as	O
antitumor	B-Cancer
agents	O
in	O
suppressing	O
tumor	B-Cancer
growth	O
and	O
metastatic	O
progression	O
.	O

Vasostatin	B-Gene_or_gene_product
,	O
the	O
N	O
-	O
terminal	O
domain	O
of	O
calreticulin	B-Gene_or_gene_product
,	O
is	O
a	O
potent	O
angiogenesis	O
inhibitor	O
.	O

In	O
this	O
study	O
,	O
we	O
determined	O
the	O
effectiveness	O
of	O
vasostatin	B-Gene_or_gene_product
delivered	O
by	O
recombinant	B-Organism
pseudotype	I-Organism
adeno	I-Organism
-	I-Organism
associated	I-Organism
virus	I-Organism
2	I-Organism
/	O
5	B-Organism
(	O
rAAV2	B-Organism
/	O
5	B-Organism
-	I-Organism
VAS	I-Organism
)	O
as	O
a	O
gene	O
therapy	O
approach	O
for	O
lung	B-Cancer
cancer	I-Cancer
treatment	O
.	O

EXPERIMENTAL	O
DESIGN	O
:	O
We	O
used	O
rAAV2	B-Organism
/	O
5	B-Organism
to	O
deliver	O
vasostatin	B-Gene_or_gene_product
intratumorally	B-Cancer
or	O
systemically	O
in	O
different	O
mouse	B-Organism
lung	B-Cancer
tumor	I-Cancer
models	O
-	O
-	O
subcutaneous	O
,	O
orthotopic	O
xenograft	B-Cancer
,	O
and	O
spontaneous	O
metastasis	O
lung	B-Cancer
tumor	I-Cancer
models	O
.	O

The	O
therapeutic	O
efficacy	O
of	O
rAAV2	B-Organism
/	O
5	B-Organism
-	I-Organism
VAS	I-Organism
was	O
determined	O
by	O
monitoring	O
tumor	B-Cancer
volume	O
,	O
survival	O
rate	O
,	O
and	O
degree	O
of	O
neovascularization	O
after	O
treatment	O
in	O
these	O
models	O
.	O

RESULTS	O
:	O
Mice	B-Organism
bearing	O
subcutaneous	B-Cancer
tumor	I-Cancer
of	O
rAAV2	B-Organism
/	O
5	B-Organism
-	I-Organism
VAS	I-Organism
pretreated	O
Lewis	B-Cell
lung	I-Cell
carcinoma	I-Cell
cells	I-Cell
showed	O
>	O
50	O
%	O
reduction	O
in	O
primary	O
tumor	B-Cancer
volume	O
and	O
reduced	O
spontaneous	O
pulmonary	B-Organ
metastases	O
.	O

The	O
tumor	B-Cancer
-	O
suppressive	O
action	O
of	O
rAAV2	B-Organism
/	O
5	B-Organism
-	I-Organism
VAS	I-Organism
in	O
subcutaneous	O
human	B-Organism
lung	B-Cancer
tumor	I-Cancer
A549	I-Cancer
xenograft	I-Cancer
correlated	O
with	O
a	O
reduced	O
number	O
of	O
capillary	B-Tissue
vessels	I-Tissue
in	O
tumors	B-Cancer
.	O

In	O
the	O
orthotopic	O
xenograft	B-Cancer
model	O
,	O
rAAV2	B-Organism
/	O
5	B-Organism
-	I-Organism
VAS	I-Organism
suppressed	O
metastasis	O
of	O
A549	B-Cancer
tumors	I-Cancer
to	O
mediastinal	O
lymph	B-Multi-tissue_structure
nodes	I-Multi-tissue_structure
and	O
contralateral	O
lung	B-Organ
.	O

Furthermore	O
,	O
treatment	O
of	O
immunocompetent	O
mice	B-Organism
in	O
the	O
spontaneous	O
lung	B-Organ
metastases	O
model	O
with	O
rAAV2	B-Organism
/	O
5	B-Organism
-	I-Organism
VAS	I-Organism
after	O
primary	O
tumor	B-Cancer
excision	O
prolonged	O
their	O
median	O
survival	O
from	O
21	O
to	O
51	O
.	O
5	O
days	O
.	O

CONCLUSION	O
:	O
Our	O
results	O
show	O
the	O
effectiveness	O
of	O
rAAV2	B-Organism
/	O
5	B-Organism
-	I-Organism
VAS	I-Organism
as	O
an	O
angiogenesis	O
inhibitor	O
in	O
suppressing	O
tumor	B-Cancer
growth	O
during	O
different	O
stages	O
of	O
tumor	B-Cancer
progression	O
,	O
validating	O
the	O
application	O
of	O
rAAV2	B-Organism
/	O
5	B-Organism
-	I-Organism
VAS	I-Organism
gene	O
therapy	O
in	O
treatment	O
against	O
lung	B-Cancer
cancer	I-Cancer
.	O

Radiation	O
retinopathy	O
is	O
treatable	O
with	O
anti	B-Gene_or_gene_product
-	I-Gene_or_gene_product
vascular	I-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
bevacizumab	B-Simple_chemical
(	O
Avastin	B-Simple_chemical
)	O
.	O

PURPOSE	O
:	O
To	O
report	O
on	O
bevacizumab	B-Simple_chemical
treatment	O
for	O
radiation	O
retinopathy	O
affecting	O
the	O
macula	B-Tissue
.	O

PATIENTS	O
AND	O
METHODS	O
:	O
Twenty	O
-	O
one	O
patients	B-Organism
with	O
radiation	O
retinopathy	O
(	O
edema	B-Pathological_formation
,	O
hemorrhages	O
,	O
capillary	B-Tissue
dropout	O
,	O
and	O
neovascularization	O
)	O
and	O
a	O
subjective	O
or	O
objective	O
loss	O
of	O
vision	O
were	O
treated	O
.	O

Treatment	O
involved	O
intravitreal	O
injection	O
of	O
bevacizumab	B-Simple_chemical
(	O
1	O
.	O
25	O
mg	O
in	O
0	O
.	O
05	O
mL	O
)	O
every	O
6	O
-	O
12	O
weeks	O
.	O

Treatment	O
was	O
discontinued	O
at	O
patient	B-Organism
request	O
or	O
if	O
there	O
was	O
no	O
measurable	O
response	O
to	O
therapy	O
.	O

Main	O
outcome	O
measures	O
included	O
best	O
corrected	O
visual	O
acuity	O
,	O
ophthalmic	O
examination	O
,	O
retinal	B-Multi-tissue_structure
photography	O
,	O
and	O
angiography	O
.	O

RESULTS	O
:	O
Bevacizumab	B-Simple_chemical
treatment	O
was	O
followed	O
by	O
reductions	O
in	O
retinal	B-Multi-tissue_structure
hemorrhage	O
,	O
exudation	O
,	O
and	O
edema	B-Pathological_formation
.	O

Visual	O
acuities	O
were	O
stable	O
or	O
improved	O
in	O
86	O
%	O
(	O
n	O
=	O
18	O
)	O
.	O

Three	O
patients	B-Organism
discontinued	O
therapy	O
.	O

Each	O
was	O
legally	O
blind	O
before	O
treatment	O
(	O
n	O
=	O
1	O
)	O
,	O
experienced	O
little	O
to	O
no	O
subjective	O
improvement	O
(	O
n	O
=	O
2	O
)	O
,	O
or	O
was	O
poorly	O
compliant	O
(	O
n	O
=	O
2	O
)	O
.	O

Three	O
patients	B-Organism
(	O
14	O
%	O
)	O
regained	O
2	O
or	O
more	O
lines	O
of	O
visual	O
acuity	O
.	O

No	O
ocular	O
or	O
systemic	O
bevacizumab	B-Simple_chemical
-	O
related	O
side	O
effects	O
were	O
observed	O
.	O

CONCLUSIONS	O
:	O
Intravitreal	O
bevacizumab	B-Simple_chemical
can	O
be	O
used	O
to	O
treat	O
radiation	O
retinopathy	O
.	O

In	O
most	O
cases	O
treatment	O
was	O
associated	O
with	O
decreased	O
vascular	B-Multi-tissue_structure
leakage	O
,	O
stabilization	O
,	O
or	O
improved	O
vision	O
.	O

An	O
anti	O
-	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
strategy	O
may	O
reduce	O
tissue	B-Tissue
damage	O
associated	O
with	O
radiation	O
vasculopathy	O
and	O
neuropathy	O
.	O

Mitogen	B-Gene_or_gene_product
-	I-Gene_or_gene_product
activated	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
signaling	O
promotes	O
growth	O
and	O
vascularization	O
of	O
fibrosarcoma	B-Cancer
.	O

We	O
hypothesized	O
that	O
signaling	O
through	O
multiple	O
mitogen	B-Gene_or_gene_product
-	I-Gene_or_gene_product
activated	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
(	I-Gene_or_gene_product
MAPK	I-Gene_or_gene_product
)	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
(	O
MKK	B-Gene_or_gene_product
)	O
pathways	O
is	O
essential	O
for	O
the	O
growth	O
and	O
vascularization	O
of	O
soft	B-Cancer
-	I-Cancer
tissue	I-Cancer
sarcomas	I-Cancer
,	O
which	O
are	O
malignant	B-Cancer
tumors	I-Cancer
derived	O
from	O
mesenchymal	B-Tissue
tissues	I-Tissue
.	O

We	O
tested	O
this	O
using	O
HT	B-Cell
-	I-Cell
1080	I-Cell
,	O
NCI	B-Cell
,	O
and	O
Shac	B-Cell
fibrosarcoma	I-Cell
-	I-Cell
derived	I-Cell
cell	I-Cell
lines	I-Cell
and	O
anthrax	B-Organism
lethal	B-Simple_chemical
toxin	I-Simple_chemical
(	O
LeTx	B-Simple_chemical
)	O
,	O
a	O
bacterial	O
toxin	O
that	O
inactivates	O
MKKs	B-Gene_or_gene_product
.	O

Western	O
blots	O
confirmed	O
that	O
LeTx	B-Simple_chemical
treatment	O
reduced	O
the	O
levels	O
of	O
phosphorylated	O
extracellular	B-Gene_or_gene_product
signal	I-Gene_or_gene_product
-	I-Gene_or_gene_product
regulated	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
and	O
p38	B-Gene_or_gene_product
MAPK	I-Gene_or_gene_product
in	O
vitro	O
.	O

Although	O
short	O
treatments	O
with	O
LeTx	B-Simple_chemical
only	O
modestly	O
affected	O
cell	B-Cell
proliferation	O
,	O
sustained	O
treatment	O
markedly	O
reduced	O
cell	B-Cell
numbers	O
.	O

LeTx	B-Simple_chemical
also	O
substantially	O
inhibited	O
the	O
extracellular	B-Immaterial_anatomical_entity
release	O
of	O
angioproliferative	O
factors	O
including	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
,	O
interleukin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
,	O
and	O
basic	B-Gene_or_gene_product
fibroblast	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
.	O

Similar	O
results	O
were	O
obtained	O
with	O
cell	B-Cell
lines	I-Cell
derived	O
from	O
malignant	B-Cancer
fibrous	I-Cancer
histiocytomas	I-Cancer
,	O
leiomyosarcomas	B-Cancer
,	O
and	O
liposarcomas	B-Cancer
.	O

In	O
vivo	O
,	O
LeTx	B-Simple_chemical
decreased	O
MAPK	B-Gene_or_gene_product
activity	O
and	O
blocked	O
fibrosarcoma	B-Cancer
growth	O
.	O

Growth	O
inhibition	O
correlated	O
with	O
decreased	O
cellular	B-Cell
proliferation	O
and	O
extensive	O
necrosis	O
,	O
and	O
it	O
was	O
accompanied	O
by	O
a	O
decrease	O
in	O
tumor	B-Cancer
mean	O
vessel	B-Multi-tissue_structure
density	O
as	O
well	O
as	O
a	O
reduction	O
in	O
serum	B-Organism_substance
expression	O
of	O
angioproliferative	O
cytokines	O
.	O

Vital	O
imaging	O
using	O
high	O
-	O
resolution	O
ultrasound	O
enhanced	O
with	O
contrast	O
microbubbles	O
revealed	O
that	O
the	O
effects	O
of	O
LeTx	B-Simple_chemical
on	O
tumor	B-Cancer
perfusion	O
were	O
remarkably	O
rapid	O
(	O
less	O
than	O
24	O
h	O
)	O
and	O
resulted	O
in	O
a	O
marked	O
reduction	O
of	O
perfusion	O
within	O
the	O
tumor	B-Cancer
but	O
not	O
in	O
nontumor	B-Tissue
tissues	I-Tissue
.	O

These	O
results	O
are	O
consistent	O
with	O
our	O
initial	O
hypothesis	O
and	O
lead	O
us	O
to	O
propose	O
that	O
MKK	B-Gene_or_gene_product
inhibition	O
by	O
LeTx	B-Simple_chemical
is	O
a	O
broadly	O
effective	O
strategy	O
for	O
targeting	O
neovascularization	O
in	O
fibrosarcomas	B-Cancer
and	O
other	O
similar	O
proliferative	B-Cancer
lesions	I-Cancer
.	O

A	O
novel	O
putative	O
tyrosine	B-Amino_acid
kinase	O
receptor	O
with	O
oncogenic	O
potential	O
.	O

We	O
have	O
detected	O
transforming	O
activity	O
by	O
a	O
tumorigenicity	O
assay	O
using	O
NIH3T3	B-Cell
cells	I-Cell
transfected	O
with	O
DNA	B-Cellular_component
from	O
a	O
chronic	O
myeloproliferative	O
disorder	O
patient	B-Organism
.	O

Here	O
,	O
we	O
report	O
the	O
cDNA	O
cloning	O
of	O
the	O
corresponding	O
oncogene	O
,	O
designated	O
UFO	B-Gene_or_gene_product
,	O
in	O
allusion	O
to	O
the	O
as	O
yet	O
unidentified	O
function	O
of	O
its	O
protein	O
.	O

Nucleotide	O
sequence	O
analysis	O
of	O
a	O
3116bp	O
cDNA	O
clone	O
revealed	O
a	O
2682	O
-	O
bp	O
-	O
long	O
open	O
reading	O
frame	O
capable	O
of	O
directing	O
the	O
synthesis	O
of	O
a	O
894	O
amino	O
acid	O
polypeptide	O
.	O

The	O
predicted	O
UFO	B-Gene_or_gene_product
protein	O
exhibits	O
characteristic	O
features	O
of	O
a	O
transmembrane	B-Cellular_component
receptor	O
with	O
associated	O
tyrosine	B-Amino_acid
kinase	O
activity	O
.	O

The	O
UFO	B-Gene_or_gene_product
proto	O
-	O
oncogene	O
maps	O
to	O
human	B-Organism
chromosome	O
19q13	O
.	O
1	O
and	O
is	O
transcribed	O
into	O
two	O
5	O
.	O
0	O
kb	O
and	O
3	O
.	O
2	O
kb	O
mRNAs	O
in	O
human	B-Organism
bone	B-Multi-tissue_structure
marrow	I-Multi-tissue_structure
and	O
human	B-Organism
tumor	B-Cell
cell	I-Cell
lines	I-Cell
.	O

The	O
UFO	B-Gene_or_gene_product
locus	O
is	O
evolutionarily	O
conserved	O
between	O
vertebrate	O
species	O
.	O

A	O
4	O
.	O
0	O
kb	O
mRNA	O
of	O
the	O
murine	B-Organism
UFO	B-Gene_or_gene_product
homolog	O
is	O
expressed	O
in	O
a	O
variety	O
of	O
different	O
mouse	B-Organism
tissues	B-Tissue
.	O

We	O
thus	O
have	O
identified	O
a	O
novel	O
element	O
of	O
the	O
complex	O
signaling	O
network	O
involved	O
in	O
the	O
control	O
of	O
cell	B-Cell
proliferation	O
and	O
differentiation	O
.	O

Proteomic	O
comparison	O
of	O
nasopharyngeal	B-Cell
cancer	I-Cell
cell	I-Cell
lines	I-Cell
C666	I-Cell
-	I-Cell
1	I-Cell
and	O
NP69	B-Cell
identifies	O
down	O
-	O
regulation	O
of	O
annexin	B-Gene_or_gene_product
II	I-Gene_or_gene_product
and	O
beta2	B-Gene_or_gene_product
-	I-Gene_or_gene_product
tubulin	I-Gene_or_gene_product
for	O
nasopharyngeal	B-Cancer
carcinoma	I-Cancer
.	O

CONTEXT	O
:	O
Nasopharyngeal	B-Cancer
carcinoma	I-Cancer
(	O
NPC	B-Cancer
)	O
,	O
common	O
in	O
southern	O
China	O
and	O
North	O
Africa	O
,	O
has	O
a	O
complex	O
etiology	O
involving	O
interplay	O
between	O
viral	O
,	O
environmental	O
,	O
and	O
hereditary	O
factors	O
and	O
is	O
almost	O
constantly	O
associated	O
with	O
the	O
Epstein	B-Organism
-	I-Organism
Barr	I-Organism
virus	I-Organism
.	O

Since	O
the	O
prognosis	O
of	O
locally	O
advanced	O
and	O
metastatic	O
diseases	O
is	O
poor	O
,	O
increased	O
understanding	O
of	O
the	O
pathogenesis	O
of	O
NPC	B-Cancer
would	O
be	O
important	O
for	O
discovering	O
novel	O
markers	O
for	O
patients	B-Organism
'	O
management	O
.	O

OBJECTIVES	O
:	O
To	O
compare	O
the	O
proteomic	O
expression	O
profile	O
between	O
an	O
Epstein	B-Organism
-	I-Organism
Barr	I-Organism
virus	I-Organism
-	O
associated	O
NPC	B-Cell
cell	I-Cell
line	I-Cell
(	O
C666	B-Cell
-	I-Cell
1	I-Cell
)	O
and	O
a	O
normal	O
NP	B-Cell
cell	I-Cell
line	I-Cell
(	O
NP69	B-Cell
)	O
.	O

The	O
proteins	O
with	O
differential	O
expression	O
were	O
analyzed	O
in	O
40	O
undifferentiated	O
NPC	B-Cell
paraffin	O
-	O
embedded	O
specimens	B-Tissue
.	O

DESIGN	O
:	O
Differentially	O
expressed	O
proteins	O
discovered	O
between	O
the	O
two	O
cell	B-Cell
lines	I-Cell
were	O
identified	O
by	O
mass	O
spectrometry	O
.	O

After	O
confirmation	O
by	O
immunocytochemical	O
staining	O
,	O
their	O
expression	O
in	O
patient	B-Organism
samples	O
was	O
measured	O
using	O
40	O
pairs	O
of	O
undifferentiated	O
NPCs	B-Cell
together	O
with	O
their	O
adjacent	O
normal	B-Tissue
epithelia	I-Tissue
.	O

RESULTS	O
:	O
Proteomic	O
findings	O
indicated	O
that	O
adenosine	B-Gene_or_gene_product
triphosphate	I-Gene_or_gene_product
synthase	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
chain	I-Gene_or_gene_product
was	O
up	O
-	O
regulated	O
,	O
whereas	O
annexin	B-Gene_or_gene_product
II	I-Gene_or_gene_product
,	O
annexin	B-Gene_or_gene_product
V	I-Gene_or_gene_product
,	O
beta	B-Gene_or_gene_product
(	I-Gene_or_gene_product
2	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	I-Gene_or_gene_product
tubulin	I-Gene_or_gene_product
,	O
and	O
profilin	B-Gene_or_gene_product
1	I-Gene_or_gene_product
were	O
down	O
-	O
regulated	O
.	O

After	O
confirming	O
the	O
results	O
in	O
agar	O
-	O
processed	O
cell	B-Cell
lines	I-Cell
,	O
annexin	B-Gene_or_gene_product
II	I-Gene_or_gene_product
and	O
beta	B-Gene_or_gene_product
(	I-Gene_or_gene_product
2	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	I-Gene_or_gene_product
tubulin	I-Gene_or_gene_product
expression	O
were	O
found	O
to	O
be	O
lower	O
in	O
tumor	B-Cell
cells	I-Cell
than	O
in	O
adjacent	O
normal	B-Cell
epithelial	I-Cell
cells	I-Cell
in	O
100	O
%	O
and	O
90	O
%	O
of	O
the	O
patients	B-Organism
'	O
specimens	B-Cell
,	O
respectively	O
.	O

Finally	O
,	O
annexin	B-Gene_or_gene_product
II	I-Gene_or_gene_product
down	O
-	O
regulation	O
was	O
positively	O
associated	O
with	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
,	O
suggesting	O
that	O
it	O
may	O
be	O
a	O
prognostic	O
factor	O
in	O
NPC	B-Cancer
.	O

CONCLUSIONS	O
:	O
The	O
results	O
suggest	O
that	O
annexin	B-Gene_or_gene_product
II	I-Gene_or_gene_product
and	O
beta	B-Gene_or_gene_product
(	I-Gene_or_gene_product
2	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	I-Gene_or_gene_product
tubulin	I-Gene_or_gene_product
down	O
-	O
regulation	O
is	O
important	O
in	O
NPC	B-Cancer
formation	O
and	O
may	O
represent	O
potential	O
targets	O
for	O
further	O
investigations	O
.	O

MMTV	B-Organism
-	O
cre	B-Gene_or_gene_product
-	O
mediated	O
fur	B-Gene_or_gene_product
inactivation	O
concomitant	O
with	O
PLAG1	B-Gene_or_gene_product
proto	O
-	O
oncogene	O
activation	O
delays	O
salivary	B-Multi-tissue_structure
gland	I-Multi-tissue_structure
tumorigenesis	O
in	O
mice	B-Organism
.	O

Proprotein	O
convertases	O
are	O
serine	B-Amino_acid
endoproteases	O
implicated	O
in	O
the	O
proteolytic	O
processing	O
of	O
a	O
large	O
variety	O
of	O
regulatory	O
proteins	O
.	O

An	O
important	O
role	O
of	O
proprotein	O
convertases	O
in	O
tumorigenic	O
processes	O
has	O
been	O
suggested	O
by	O
various	O
studies	O
.	O

In	O
this	O
study	O
,	O
the	O
role	O
of	O
the	O
proprotein	O
convertase	O
furin	B-Gene_or_gene_product
in	O
PLAG1	B-Gene_or_gene_product
proto	O
-	O
oncogene	O
-	O
induced	O
salivary	B-Multi-tissue_structure
gland	I-Multi-tissue_structure
tumorigenesis	O
was	O
investigated	O
.	O

PLAG1	B-Gene_or_gene_product
overexpression	O
in	O
salivary	B-Organ
glands	I-Organ
has	O
previously	O
been	O
shown	O
to	O
result	O
in	O
salivary	B-Cancer
gland	I-Cancer
tumors	I-Cancer
in	O
100	O
%	O
of	O
mice	B-Organism
within	O
5	O
weeks	O
after	O
birth	O
.	O

MMTV	B-Organism
-	O
cre	B-Gene_or_gene_product
-	O
mediated	O
inactivation	O
of	O
fur	B-Gene_or_gene_product
without	O
over	O
-	O
expression	O
of	O
PLAG1	B-Gene_or_gene_product
caused	O
smaller	O
but	O
histologically	O
normal	O
salivary	B-Organ
glands	I-Organ
.	O

Moreover	O
,	O
the	O
lymph	B-Multi-tissue_structure
nodes	I-Multi-tissue_structure
close	O
to	O
the	O
salivary	B-Organ
glands	I-Organ
were	O
enlarged	O
,	O
and	O
histology	O
showed	O
that	O
they	O
had	O
activated	O
follicles	B-Multi-tissue_structure
.	O

When	O
genetic	O
ablation	O
of	O
1	O
or	O
2	O
alleles	O
of	O
fur	B-Gene_or_gene_product
and	O
overexpression	O
of	O
the	O
PLAG1	B-Gene_or_gene_product
transgene	O
were	O
simultaneously	O
achieved	O
,	O
a	O
significant	O
delay	O
in	O
tumorigenesis	O
was	O
observed	O
.	O

Collectively	O
,	O
these	O
results	O
suggest	O
an	O
important	O
role	O
for	O
furin	B-Gene_or_gene_product
in	O
PLAG1	B-Gene_or_gene_product
-	O
induced	O
salivary	B-Multi-tissue_structure
gland	I-Multi-tissue_structure
tumorigenesis	O
in	O
mice	B-Organism
.	O

Isotype	O
-	O
specific	O
inhibitors	O
of	O
the	O
glycolytic	O
key	O
regulator	O
pyruvate	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
subtype	I-Gene_or_gene_product
M2	I-Gene_or_gene_product
moderately	O
decelerate	O
tumor	B-Cell
cell	I-Cell
proliferation	O
.	O

Tumor	B-Cell
cells	I-Cell
express	O
the	O
glycolytic	O
regulator	O
pyruvate	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
subtype	I-Gene_or_gene_product
M2	I-Gene_or_gene_product
(	O
M2	B-Gene_or_gene_product
-	I-Gene_or_gene_product
PK	I-Gene_or_gene_product
)	O
,	O
which	O
can	O
occur	O
in	O
a	O
tetrameric	O
form	O
with	O
high	O
affinity	O
to	O
its	O
substrate	O
phosphoenolpyruvate	B-Simple_chemical
(	O
PEP	B-Simple_chemical
)	O
and	O
a	O
dimeric	O
form	O
with	O
a	O
low	O
PEP	B-Simple_chemical
affinity	O
.	O

The	O
transition	O
between	O
both	O
conformations	O
contributes	O
to	O
the	O
control	O
of	O
glycolysis	O
and	O
is	O
important	O
for	O
tumor	B-Cell
cell	I-Cell
proliferation	O
and	O
survival	O
.	O

Here	O
we	O
targeted	O
M2	B-Gene_or_gene_product
-	I-Gene_or_gene_product
PK	I-Gene_or_gene_product
by	O
synthetic	O
peptide	O
aptamers	O
,	O
which	O
specifically	O
bind	O
to	O
M2	B-Gene_or_gene_product
-	I-Gene_or_gene_product
PK	I-Gene_or_gene_product
and	O
shift	O
the	O
isoenzyme	O
into	O
its	O
low	O
affinity	O
dimeric	O
conformation	O
.	O

The	O
aptamer	O
-	O
induced	O
dimerization	O
and	O
inactivation	O
of	O
M2	B-Gene_or_gene_product
-	I-Gene_or_gene_product
PK	I-Gene_or_gene_product
led	O
to	O
a	O
significant	O
decrease	O
in	O
the	O
PK	B-Gene_or_gene_product
mass	O
-	O
action	O
ratio	O
as	O
well	O
as	O
ATP	B-Simple_chemical
:	O
ADP	B-Simple_chemical
ratio	O
in	O
the	O
target	O
cells	B-Cell
.	O

Furthermore	O
,	O
the	O
expression	O
of	O
M2	B-Gene_or_gene_product
-	I-Gene_or_gene_product
PK	I-Gene_or_gene_product
-	O
binding	O
peptide	O
aptamers	O
moderately	O
reduced	O
the	O
growth	O
of	O
immortalized	O
NIH3T3	B-Cell
cell	I-Cell
populations	I-Cell
by	O
decelerating	O
cell	B-Cell
proliferation	O
,	O
but	O
without	O
affecting	O
apoptotic	O
cell	B-Cell
death	O
.	O

Moreover	O
,	O
the	O
M2	B-Gene_or_gene_product
-	I-Gene_or_gene_product
PK	I-Gene_or_gene_product
-	O
binding	O
peptide	O
aptamers	O
also	O
reduced	O
the	O
proliferation	O
rate	O
of	O
human	B-Organism
U	B-Cell
-	I-Cell
2	I-Cell
OS	I-Cell
osteosarcoma	I-Cell
cells	I-Cell
.	O

In	O
the	O
present	O
study	O
,	O
we	O
developed	O
the	O
first	O
specific	O
inhibitors	O
of	O
the	O
pyruvate	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
isoenzyme	I-Gene_or_gene_product
type	I-Gene_or_gene_product
M2	I-Gene_or_gene_product
and	O
present	O
evidence	O
that	O
these	O
inhibitors	O
moderately	O
decelerate	O
tumor	B-Cell
cell	I-Cell
proliferation	O
.	O

Hematogenous	O
metastasis	O
in	O
gastric	B-Cancer
cancer	I-Cancer
requires	O
isolated	B-Cell
tumor	I-Cell
cells	I-Cell
and	O
expression	O
of	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

PURPOSE	O
:	O
Recent	O
studies	O
of	O
cancer	B-Cancer
metastasis	O
have	O
focused	O
on	O
the	O
role	O
of	O
premetastatic	O
gene	O
expression	O
and	O
circulating	O
tumor	B-Cell
cells	I-Cell
.	O

We	O
did	O
a	O
blind	O
prospective	O
study	O
in	O
gastric	B-Cancer
cancer	I-Cancer
to	O
assess	O
the	O
significance	O
of	O
isolated	B-Cell
tumor	I-Cell
cells	I-Cell
(	O
ITC	B-Cell
)	O
and	O
to	O
test	O
the	O
hypothesis	O
that	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
is	O
expressed	O
within	O
the	O
bone	B-Multi-tissue_structure
marrow	I-Multi-tissue_structure
at	O
tumor	B-Cancer
-	O
specific	O
,	O
premetastatic	B-Multi-tissue_structure
sites	I-Multi-tissue_structure
.	O

EXPERIMENTAL	O
DESIGN	O
:	O
Both	O
bone	B-Multi-tissue_structure
marrow	I-Multi-tissue_structure
and	O
peripheral	B-Organism_substance
blood	I-Organism_substance
samples	I-Organism_substance
from	O
810	O
gastric	B-Cancer
cancer	I-Cancer
patients	B-Organism
were	O
collected	O
at	O
the	O
Central	O
Hospital	O
,	O
National	O
Cancer	O
Center	O
(	O
Tokyo	O
,	O
Japan	O
)	O
.	O

The	O
samples	O
were	O
transferred	O
to	O
Kyushu	O
University	O
Hospital	O
(	O
Beppu	O
,	O
Japan	O
)	O
where	O
they	O
were	O
analyzed	O
by	O
quantitative	O
real	O
-	O
time	O
reverse	O
transcription	O
-	O
PCR	O
for	O
three	O
epithelial	B-Cell
cell	I-Cell
markers	O
,	O
carcinoembryonic	B-Gene_or_gene_product
antigen	I-Gene_or_gene_product
,	O
cytokeratin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
19	I-Gene_or_gene_product
,	O
and	O
cytokeratin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
7	I-Gene_or_gene_product
,	O
as	O
well	O
as	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

RESULTS	O
:	O
ITCs	B-Cell
were	O
observed	O
in	O
peripheral	B-Organism_substance
blood	I-Organism_substance
and	O
bone	B-Multi-tissue_structure
marrow	I-Multi-tissue_structure
even	O
in	O
early	O
stages	O
of	O
gastric	B-Cancer
cancer	I-Cancer
.	O

The	O
frequency	O
of	O
ITC	B-Cell
in	O
bone	B-Multi-tissue_structure
marrow	I-Multi-tissue_structure
was	O
significantly	O
associated	O
with	O
the	O
stage	O
of	O
disease	O
by	O
ANOVA	O
(	O
P	O
<	O
0	O
.	O
01	O
)	O
.	O

Gastric	B-Cancer
cancer	I-Cancer
metastasized	O
when	O
ITCs	B-Cell
were	O
observed	O
in	O
the	O
presence	O
of	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

In	O
the	O
380	O
patients	B-Organism
who	O
were	O
ITC	B-Cell
negative	O
and	O
showed	O
low	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
expression	O
,	O
synchronous	O
(	O
at	O
the	O
time	O
of	O
surgery	O
)	O
and	O
heterochronous	O
(	O
recurrent	O
)	O
metastases	O
were	O
not	O
observed	O
.	O

CONCLUSIONS	O
:	O
ITCs	B-Cell
circulate	O
even	O
in	O
early	O
stages	O
of	O
disease	O
.	O

Furthermore	O
,	O
elevated	O
expression	O
of	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
facilitates	O
the	O
establishment	O
of	O
hematogenous	O
metastases	O
in	O
gastric	B-Cancer
cancer	I-Cancer
.	O

This	O
study	O
indicates	O
that	O
the	O
simultaneous	O
presence	O
of	O
ITC	B-Cell
and	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
expression	O
at	O
premetastatic	B-Multi-tissue_structure
sites	I-Multi-tissue_structure
is	O
clinically	O
significant	O
for	O
disease	O
progression	O
.	O

Acute	O
ethanol	B-Simple_chemical
exposure	O
disrupts	O
VEGF	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
cell	B-Cell
signaling	O
in	O
endothelial	B-Cell
cells	I-Cell
.	O

Physiological	O
angiogenesis	O
is	O
regulated	O
by	O
various	O
factors	O
,	O
including	O
signaling	O
through	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	I-Gene_or_gene_product
VEGF	I-Gene_or_gene_product
)	I-Gene_or_gene_product
receptors	I-Gene_or_gene_product
.	O

We	O
previously	O
reported	O
that	O
a	O
single	O
dose	O
of	O
ethanol	B-Simple_chemical
(	O
1	O
.	O
4	O
g	O
/	O
kg	O
)	O
,	O
yielding	O
a	O
blood	B-Organism_substance
alcohol	B-Simple_chemical
concentration	O
of	O
100	O
mg	O
/	O
dl	O
,	O
significantly	O
impairs	O
angiogenesis	O
in	O
murine	B-Organism
wounds	B-Pathological_formation
,	O
despite	O
adequate	O
levels	O
of	O
VEGF	B-Gene_or_gene_product
,	O
suggesting	O
direct	O
effects	O
of	O
ethanol	B-Simple_chemical
on	O
endothelial	B-Cell
cell	I-Cell
signaling	O
(	O
40	O
)	O
.	O

To	O
examine	O
the	O
mechanism	O
by	O
which	O
ethanol	B-Simple_chemical
influences	O
angiogenesis	O
in	O
wounds	B-Pathological_formation
,	O
we	O
employed	O
two	O
different	O
in	O
vitro	O
angiogenesis	O
assays	O
to	O
determine	O
whether	O
acute	O
ethanol	B-Simple_chemical
exposure	O
(	O
100	O
mg	O
/	O
dl	O
)	O
would	O
have	O
long	O
-	O
lasting	O
effects	O
on	O
VEGF	B-Gene_or_gene_product
-	O
induced	O
capillary	B-Multi-tissue_structure
network	I-Multi-tissue_structure
formation	O
.	O

Ethanol	B-Simple_chemical
exposure	O
resulted	O
in	O
reduced	O
VEGF	B-Gene_or_gene_product
-	O
induced	O
cord	B-Tissue
formation	O
on	O
collagen	B-Gene_or_gene_product
and	O
reduced	O
capillary	B-Multi-tissue_structure
network	I-Multi-tissue_structure
structure	O
on	O
Matrigel	O
in	O
vitro	O
.	O

In	O
addition	O
,	O
ethanol	B-Simple_chemical
exposure	O
decreased	O
expression	O
of	O
endothelial	B-Cell
VEGF	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
as	O
well	O
as	O
VEGF	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
phosphorylation	O
in	O
vitro	O
.	O

Inhibition	O
of	O
ethanol	B-Simple_chemical
metabolism	O
by	O
4	B-Simple_chemical
-	I-Simple_chemical
methylpyrazole	I-Simple_chemical
partially	O
abrogated	O
the	O
effect	O
of	O
ethanol	B-Simple_chemical
on	O
endothelial	B-Tissue
cell	I-Tissue
cord	I-Tissue
formation	O
.	O

However	O
,	O
mice	B-Organism
treated	O
with	O
t	B-Simple_chemical
-	I-Simple_chemical
butanol	I-Simple_chemical
,	O
an	O
alcohol	B-Simple_chemical
not	O
metabolized	O
by	O
alcohol	B-Gene_or_gene_product
dehydrogenase	I-Gene_or_gene_product
,	O
exhibited	O
no	O
change	O
in	O
wound	B-Pathological_formation
vascularity	O
.	O

These	O
results	O
suggest	O
that	O
products	O
of	O
ethanol	B-Simple_chemical
metabolism	O
are	O
important	O
factors	O
in	O
the	O
development	O
of	O
ethanol	B-Simple_chemical
-	O
induced	O
changes	O
in	O
endothelial	B-Cell
cell	I-Cell
responsiveness	O
to	O
VEGF	B-Gene_or_gene_product
.	O

In	O
vivo	O
,	O
ethanol	B-Simple_chemical
exposure	O
caused	O
both	O
decreased	O
angiogenesis	O
and	O
increased	O
hypoxia	O
in	O
wounds	B-Pathological_formation
.	O

Moreover	O
,	O
in	O
vitro	O
experiments	O
demonstrated	O
a	O
direct	O
effect	O
of	O
ethanol	B-Simple_chemical
on	O
the	O
response	O
to	O
hypoxia	O
in	O
endothelial	B-Cell
cells	I-Cell
,	O
as	O
ethanol	B-Simple_chemical
diminished	O
nuclear	O
hypoxia	B-Gene_or_gene_product
-	I-Gene_or_gene_product
inducible	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
protein	O
levels	O
.	O

Together	O
,	O
the	O
data	O
establish	O
that	O
acute	O
ethanol	B-Simple_chemical
exposure	O
significantly	O
impairs	O
angiogenesis	O
and	O
suggest	O
that	O
this	O
effect	O
is	O
mediated	O
by	O
changes	O
in	O
endothelial	B-Cell
cell	I-Cell
responsiveness	O
to	O
both	O
VEGF	B-Gene_or_gene_product
and	O
hypoxia	O
.	O

Adenomatoid	B-Cancer
tumour	I-Cancer
of	O
the	O
liver	B-Organ
.	O

An	O
unusual	O
primary	O
adenomatoid	B-Cancer
tumour	I-Cancer
arising	O
in	O
the	O
normal	O
liver	B-Organ
is	O
described	O
.	O

Hepatectomy	O
was	O
performed	O
,	O
and	O
the	O
patient	B-Organism
is	O
alive	O
and	O
free	O
of	O
disease	O
1	O
year	O
postsurgery	O
.	O

Grossly	O
,	O
the	O
tumour	B-Cancer
showed	O
a	O
haemorrhagic	O
cut	O
surface	O
with	O
numerous	O
microcystic	B-Pathological_formation
structures	I-Pathological_formation
.	O

Histological	O
examination	O
revealed	O
cystic	O
or	O
angiomatoid	O
spaces	O
of	O
various	O
sizes	O
lined	O
by	O
cuboidal	O
,	O
low	O
-	O
columnar	O
,	O
or	O
flattened	O
epithelioid	B-Cell
cells	I-Cell
with	O
vacuolated	B-Organism_substance
cytoplasm	I-Organism_substance
and	O
round	O
to	O
oval	O
nuclei	B-Cellular_component
.	O

The	O
epithelioid	B-Cell
cells	I-Cell
were	O
entirely	O
supported	O
by	O
proliferated	O
capillaries	B-Tissue
and	O
arteries	B-Multi-tissue_structure
together	O
with	O
collagenous	B-Tissue
stroma	I-Tissue
.	O

Immunohistochemical	O
studies	O
showed	O
that	O
the	O
epithelioid	B-Cell
cells	I-Cell
were	O
strongly	O
positive	O
for	O
a	O
broad	O
spectrum	O
of	O
cytokeratins	B-Gene_or_gene_product
(	O
AE1	B-Gene_or_gene_product
/	O
AE3	B-Gene_or_gene_product
,	O
CAM5	B-Gene_or_gene_product
.	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
epithelial	B-Gene_or_gene_product
membrane	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
and	O
cytokeratin	B-Gene_or_gene_product
7	I-Gene_or_gene_product
)	O
and	O
mesothelial	B-Tissue
markers	O
(	O
calretinin	B-Gene_or_gene_product
,	O
Wilms	B-Gene_or_gene_product
'	I-Gene_or_gene_product
tumour	I-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
D2	B-Gene_or_gene_product
-	I-Gene_or_gene_product
40	I-Gene_or_gene_product
)	O
.	O

These	O
cells	B-Cell
were	O
negative	O
for	O
Hep	B-Gene_or_gene_product
par	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
carcinoembryonic	B-Gene_or_gene_product
antigen	I-Gene_or_gene_product
,	O
neural	B-Gene_or_gene_product
cell	I-Gene_or_gene_product
adhesion	I-Gene_or_gene_product
molecule	I-Gene_or_gene_product
,	O
CD34	B-Gene_or_gene_product
,	O
CD31	B-Gene_or_gene_product
and	O
HMB45	B-Gene_or_gene_product
.	O

Atypically	O
,	O
abundant	O
capillaries	B-Tissue
were	O
observed	O
;	O
however	O
,	O
the	O
cystic	B-Pathological_formation
proliferation	O
of	O
epithelioid	B-Cell
cells	I-Cell
with	O
vacuoles	B-Cellular_component
and	O
immunohistochemical	O
profile	O
of	O
the	O
epithelioid	B-Tissue
element	I-Tissue
were	O
consistent	O
with	O
hepatic	B-Cancer
adenomatoid	I-Cancer
tumour	I-Cancer
.	O

Contemporary	O
perspectives	O
on	O
vital	O
pulp	B-Tissue
therapy	O
:	O
views	O
from	O
the	O
endodontists	O
and	O
pediatric	O
dentists	O
.	O

The	O
purpose	O
of	O
this	O
study	O
was	O
to	O
determine	O
the	O
level	O
of	O
agreement	O
between	O
pediatric	O
dentists	O
and	O
endodontists	O
at	O
a	O
pulp	B-Tissue
therapy	O
symposium	O
conjointly	O
sponsored	O
by	O
the	O
American	O
Association	O
of	O
Endodontists	O
(	O
AAE	O
)	O
and	O
the	O
American	O
Academy	O
of	O
Pediatric	O
Dentistry	O
(	O
AAPD	O
)	O
on	O
November	O
2	O
-	O
3	O
,	O
2007	O
.	O

Presymposium	O
and	O
postsymposium	O
tests	O
were	O
administered	O
,	O
and	O
respondent	O
answers	O
were	O
compared	O
between	O
pediatric	O
dentists	O
and	O
endodontists	O
.	O

Opinions	O
on	O
3	O
areas	O
were	O
sought	O
:	O
pulp	B-Tissue
therapy	O
for	O
cariously	O
involved	O
primary	O
teeth	B-Organ
;	O
indirect	O
pulp	B-Tissue
treatment	O
(	O
IPT	O
)	O
for	O
cariously	O
involved	O
immature	O
permanent	O
teeth	B-Organ
;	O
and	O
innovative	O
treatment	O
options	O
including	O
pulpal	B-Tissue
revascularization	O
and	O
regeneration	O
.	O

Results	O
were	O
analyzed	O
with	O
chi2	O
tests	O
.	O

Comparisons	O
of	O
presymposium	O
and	O
postsymposium	O
responses	O
and	O
between	O
the	O
2	O
groups	O
of	O
attendees	O
indicated	O
that	O
the	O
pediatric	O
dentistry	O
and	O
endodontic	O
communities	O
agree	O
that	O
formocresol	B-Simple_chemical
will	O
be	O
replaced	O
as	O
a	O
primary	O
tooth	B-Organ
pulpotomy	O
agent	O
,	O
that	O
mineral	B-Simple_chemical
trioxide	I-Simple_chemical
is	O
the	O
first	O
choice	O
to	O
take	O
its	O
place	O
,	O
that	O
IPT	O
in	O
primary	O
teeth	B-Organ
holds	O
hope	O
as	O
a	O
replacement	O
for	O
pulpotomy	O
,	O
and	O
that	O
IPT	O
is	O
an	O
acceptable	O
pulp	B-Tissue
therapy	O
technique	O
for	O
cariously	O
involved	O
young	O
permanent	O
teeth	B-Organ
.	O

Both	O
groups	O
believe	O
that	O
pulp	B-Tissue
revascularization	O
and	O
regeneration	O
will	O
be	O
viable	O
treatment	O
modalities	O
in	O
the	O
future	O
.	O

The	O
AAE	O
and	O
the	O
AAPD	O
are	O
positioned	O
to	O
begin	O
preparation	O
of	O
best	O
practice	O
guidelines	O
that	O
share	O
common	O
language	O
and	O
treatment	O
recommendations	O
for	O
pulp	B-Tissue
therapies	O
performed	O
by	O
both	O
specialties	O
.	O

The	O
role	O
of	O
platelet	B-Gene_or_gene_product
-	I-Gene_or_gene_product
derived	I-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
cell	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
/	O
thymidine	B-Gene_or_gene_product
phosphorylase	I-Gene_or_gene_product
in	O
tumor	B-Cancer
behavior	O
.	O

Platelet	B-Gene_or_gene_product
-	I-Gene_or_gene_product
derived	I-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
cell	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
-	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
PD	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ECGF	I-Gene_or_gene_product
)	O
is	O
similar	O
to	O
the	O
pyrimidine	O
enzyme	O
thymidine	B-Gene_or_gene_product
phosphorylase	I-Gene_or_gene_product
(	O
TP	B-Gene_or_gene_product
)	O
.	O

A	O
high	O
TP	B-Gene_or_gene_product
expression	O
at	O
tumor	B-Cancer
sites	O
is	O
correlated	O
with	O
tumor	B-Cancer
growth	O
,	O
induction	O
of	O
angiogenesis	O
,	O
and	O
metastasis	O
.	O

Therefore	O
,	O
high	O
TP	B-Gene_or_gene_product
is	O
most	O
likely	O
associated	O
with	O
a	O
poor	O
prognosis	O
.	O

TP	B-Gene_or_gene_product
is	O
not	O
only	O
expressed	O
in	O
tumor	B-Cell
cells	I-Cell
but	O
also	O
in	O
tumor	B-Tissue
surrounding	I-Tissue
tissues	I-Tissue
,	O
such	O
as	O
tumor	B-Cell
infiltrating	I-Cell
macrophages	I-Cell
.	O

TP	B-Gene_or_gene_product
catalyzes	O
the	O
conversion	O
of	O
thymidine	B-Simple_chemical
to	O
thymine	B-Simple_chemical
and	O
doxyribose	B-Simple_chemical
-	I-Simple_chemical
1	I-Simple_chemical
-	I-Simple_chemical
phosphate	I-Simple_chemical
(	O
dR	B-Simple_chemical
-	I-Simple_chemical
1	I-Simple_chemical
-	I-Simple_chemical
P	I-Simple_chemical
)	O
.	O

The	O
latter	O
in	O
its	O
parent	O
form	O
or	O
in	O
its	O
sugar	B-Simple_chemical
form	O
,	O
deoxyribose	B-Simple_chemical
(	O
dR	B-Simple_chemical
)	O
may	O
play	O
a	O
role	O
in	O
the	O
induction	O
of	O
angiogenesis	O
.	O

It	O
may	O
modulate	O
cellular	B-Cell
energy	O
metabolism	O
or	O
be	O
a	O
substrate	O
in	O
a	O
chemical	O
reaction	O
generating	O
reactive	B-Simple_chemical
oxygen	I-Simple_chemical
species	I-Simple_chemical
.	O

L	B-Simple_chemical
-	I-Simple_chemical
deoxyribose	I-Simple_chemical
(	O
L	B-Simple_chemical
-	I-Simple_chemical
dR	I-Simple_chemical
)	O
and	O
thymidine	B-Simple_chemical
phosphorylase	I-Simple_chemical
inhibitor	I-Simple_chemical
(	O
TPI	B-Simple_chemical
)	O
can	O
reverse	O
these	O
effects	O
.	O

The	O
mechanism	O
of	O
TP	B-Gene_or_gene_product
induction	O
is	O
not	O
yet	O
completely	O
clear	O
,	O
but	O
TNF	B-Gene_or_gene_product
,	O
IL10	B-Gene_or_gene_product
and	O
other	O
cytokines	O
have	O
been	O
clearly	O
shown	O
to	O
induce	O
its	O
expression	O
.	O

The	O
various	O
complex	O
interactions	O
of	O
TP	B-Gene_or_gene_product
give	O
it	O
an	O
essential	O
role	O
in	O
cellular	B-Cell
functioning	O
and	O
,	O
hence	O
,	O
it	O
is	O
an	O
ideal	O
target	O
in	O
cancer	B-Cancer
therapy	O
.	O

Reversal	O
of	O
the	O
malignant	O
phenotype	O
of	O
ovarian	B-Cell
cancer	I-Cell
A2780	I-Cell
cells	I-Cell
through	O
transfection	O
with	O
wild	O
-	O
type	O
PTEN	B-Gene_or_gene_product
gene	O
.	O

OBJECTIVE	O
:	O
PTEN	B-Gene_or_gene_product
(	O
phosphatase	B-Gene_or_gene_product
and	I-Gene_or_gene_product
tensin	I-Gene_or_gene_product
homologue	I-Gene_or_gene_product
deleted	I-Gene_or_gene_product
on	I-Gene_or_gene_product
chromosome	I-Gene_or_gene_product
10	I-Gene_or_gene_product
)	O
is	O
a	O
tumor	B-Cancer
suppressor	O
gene	O
identified	O
on	O
human	B-Organism
chromosome	B-Cellular_component
10q23	I-Cellular_component
.	O

Substantial	O
studies	O
have	O
demonstrated	O
that	O
PTEN	B-Gene_or_gene_product
can	O
inhibit	O
cell	B-Cell
proliferation	O
,	O
migration	O
and	O
invasion	O
of	O
many	O
cancer	B-Cell
cells	I-Cell
.	O

The	O
purpose	O
of	O
this	O
study	O
was	O
to	O
determine	O
whether	O
upregulation	O
of	O
PTEN	B-Gene_or_gene_product
gene	O
by	O
transfection	O
wild	O
-	O
type	O
PTEN	B-Gene_or_gene_product
gene	O
to	O
ovarian	B-Cell
cancer	I-Cell
cells	I-Cell
can	O
inhibit	O
growth	O
and	O
migration	O
and	O
to	O
explore	O
the	O
potential	O
for	O
PTEN	B-Gene_or_gene_product
gene	O
therapy	O
of	O
ovarian	B-Cancer
cancers	I-Cancer
.	O

METHOD	O
:	O
Wild	O
-	O
type	O
and	O
phosphatase	O
-	O
inactive	O
(	O
C124A	O
)	O
PTEN	B-Gene_or_gene_product
plasmids	O
were	O
transfected	O
into	O
ovarian	B-Cell
epithelial	I-Cell
cancer	I-Cell
A2780	I-Cell
cells	I-Cell
,	O
and	O
their	O
effects	O
on	O
cell	B-Cell
apoptosis	O
,	O
cell	B-Cell
proliferation	O
,	O
cell	B-Cell
migration	O
and	O
cell	B-Cell
invasion	O
were	O
analyzed	O
by	O
flow	O
cytometry	O
analysis	O
,	O
TUNEL	O
assay	O
,	O
MTT	O
assay	O
,	O
wound	B-Pathological_formation
-	O
healing	O
assay	O
and	O
transwell	O
assay	O
.	O

RESULTS	O
:	O
Both	O
wild	O
-	O
type	O
and	O
mutant	O
PTEN	B-Gene_or_gene_product
can	O
upregulate	O
the	O
expression	O
of	O
PTEN	B-Gene_or_gene_product
gene	O
dramatically	O
;	O
however	O
,	O
it	O
is	O
wild	O
-	O
type	O
PTEN	B-Gene_or_gene_product
not	O
phosphatase	B-Simple_chemical
-	O
inactive	O
PTEN	B-Gene_or_gene_product
that	O
can	O
induce	O
apoptosis	O
and	O
decrease	O
cell	B-Cell
migration	O
,	O
invasion	O
and	O
proliferation	O
in	O
ovarian	B-Cell
cancer	I-Cell
cells	I-Cell
.	O

CONCLUSION	O
:	O
These	O
results	O
demonstrated	O
that	O
PTEN	B-Gene_or_gene_product
had	O
played	O
an	O
important	O
role	O
in	O
the	O
cell	B-Cell
proliferation	O
,	O
cell	B-Cell
migration	O
and	O
invasion	O
dependent	O
on	O
its	O
phosphatase	O
activity	O
.	O

Enhanced	O
expression	O
of	O
PTEN	B-Gene_or_gene_product
by	O
gene	O
transfer	O
is	O
sufficient	O
to	O
reverse	O
the	O
malignant	O
phenotype	O
of	O
ovarian	B-Cell
cancer	I-Cell
cells	I-Cell
and	O
transfection	O
of	O
ovarian	B-Cell
cancer	I-Cell
cells	I-Cell
with	O
wild	O
-	O
type	O
PTEN	B-Gene_or_gene_product
gene	O
might	O
be	O
another	O
novel	O
approach	O
for	O
therapeutic	O
intervention	O
in	O
ovarian	B-Cancer
cancer	I-Cancer
.	O

Allogeneic	O
injection	O
of	O
fetal	B-Cell
membrane	I-Cell
-	I-Cell
derived	I-Cell
mesenchymal	I-Cell
stem	I-Cell
cells	I-Cell
induces	O
therapeutic	O
angiogenesis	O
in	O
a	O
rat	B-Organism
model	O
of	O
hind	B-Organism_subdivision
limb	I-Organism_subdivision
ischemia	O
.	O

Bone	B-Cell
marrow	I-Cell
-	I-Cell
derived	I-Cell
mesenchymal	I-Cell
stem	I-Cell
cells	I-Cell
(	O
BM	B-Cell
-	I-Cell
MSC	I-Cell
)	O
have	O
been	O
demonstrated	O
to	O
be	O
an	O
attractive	O
therapeutic	O
cell	B-Cell
source	O
for	O
tissue	B-Tissue
regeneration	O
and	O
repair	O
.	O

However	O
,	O
it	O
remains	O
unknown	O
whether	O
or	O
not	O
allogeneic	O
transplantation	O
of	O
mesenchymal	B-Cell
stem	I-Cell
cells	I-Cell
(	O
MSC	B-Cell
)	O
derived	O
from	O
fetal	B-Multi-tissue_structure
membranes	I-Multi-tissue_structure
(	O
FM	B-Multi-tissue_structure
)	O
,	O
which	O
are	O
generally	O
discarded	O
as	O
medical	O
waste	O
after	O
delivery	O
,	O
has	O
therapeutic	O
potential	O
.	O

FM	B-Cell
-	I-Cell
MSC	I-Cell
were	O
obtained	O
from	O
Lewis	O
rats	B-Organism
and	O
had	O
surface	O
antigen	O
expression	O
and	O
multipotent	O
potential	O
partly	O
similar	O
to	O
those	O
of	O
BM	B-Cell
-	I-Cell
MSC	I-Cell
.	O

Compared	O
with	O
BM	B-Cell
-	I-Cell
MSC	I-Cell
,	O
FM	B-Cell
-	I-Cell
MSC	I-Cell
secreted	O
a	O
comparable	O
amount	O
of	O
hepatocyte	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
despite	O
a	O
small	O
amount	O
of	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
.	O

FM	B-Cell
-	I-Cell
MSC	I-Cell
and	O
BM	B-Cell
-	I-Cell
MSC	I-Cell
both	O
expressed	O
major	O
histocompatibility	O
complex	O
(	O
MHC	O
)	O
class	O
I	O
but	O
not	O
MHC	O
class	O
II	O
antigens	O
and	O
did	O
not	O
elicit	O
allogeneic	O
lymphocyte	B-Cell
proliferation	O
in	O
mixed	O
lymphocyte	B-Cell
culture	O
.	O

FM	B-Cell
-	I-Cell
MSC	I-Cell
or	O
BM	B-Cell
-	I-Cell
MSC	I-Cell
obtained	O
from	O
Lewis	O
rats	B-Organism
were	O
injected	O
into	O
a	O
MHC	O
-	O
mismatched	O
August	O
-	O
Copenhagen	O
-	O
Irish	O
rat	B-Organism
model	O
of	O
hind	B-Organism_subdivision
limb	I-Organism_subdivision
ischemia	O
.	O

Three	O
weeks	O
after	O
injection	O
,	O
blood	B-Organism_substance
perfusion	O
and	O
capillary	B-Tissue
density	O
were	O
significantly	O
higher	O
in	O
the	O
FM	B-Cell
-	I-Cell
MSC	I-Cell
and	O
BM	B-Cell
-	I-Cell
MSC	I-Cell
groups	O
than	O
in	O
the	O
phosphate	B-Simple_chemical
-	I-Simple_chemical
buffered	I-Simple_chemical
saline	I-Simple_chemical
group	O
,	O
and	O
allogeneic	O
FM	B-Cell
-	I-Cell
MSC	I-Cell
and	O
BM	B-Cell
-	I-Cell
MSC	I-Cell
were	O
still	O
observed	O
.	O

In	O
nonischemic	O
hind	B-Tissue
limb	I-Tissue
tissues	I-Tissue
,	O
allogeneic	O
FM	B-Cell
-	I-Cell
MSC	I-Cell
and	O
BM	B-Cell
-	I-Cell
MSC	I-Cell
injection	O
were	O
associated	O
with	O
a	O
comparatively	O
small	O
amount	O
of	O
T	B-Cell
lymphocyte	I-Cell
infiltration	O
,	O
compared	O
with	O
the	O
injection	O
of	O
allogeneic	O
splenic	B-Cell
lymphocytes	I-Cell
.	O

In	O
conclusion	O
,	O
allogeneic	O
FM	B-Cell
-	I-Cell
MSC	I-Cell
injection	O
did	O
not	O
elicit	O
a	O
lymphocyte	B-Cell
proliferative	O
response	O
and	O
provided	O
significant	O
improvement	O
in	O
a	O
rat	B-Organism
model	O
of	O
hind	B-Organism_subdivision
limb	I-Organism_subdivision
ischemia	O
,	O
comparable	O
to	O
the	O
response	O
to	O
BM	B-Cell
-	I-Cell
MSC	I-Cell
.	O

Thus	O
,	O
allogeneic	O
injection	O
of	O
FM	B-Cell
-	I-Cell
MSC	I-Cell
may	O
be	O
a	O
new	O
therapeutic	O
strategy	O
for	O
the	O
treatment	O
of	O
severe	O
peripheral	B-Multi-tissue_structure
vascular	I-Multi-tissue_structure
disease	O
.	O

Disclosure	O
of	O
potential	O
conflicts	O
of	O
interest	O
is	O
found	O
at	O
the	O
end	O
of	O
this	O
article	O
.	O

Regulation	O
of	O
the	O
Warburg	O
effect	O
in	O
early	B-Cell
-	I-Cell
passage	I-Cell
breast	I-Cell
cancer	I-Cell
cells	I-Cell
.	O

Malignancy	O
in	O
cancer	B-Cancer
is	O
associated	O
with	O
aerobic	O
glycolysis	O
(	O
Warburg	O
effect	O
)	O
evidenced	O
by	O
increased	O
trapping	O
of	O
[	B-Simple_chemical
(	I-Simple_chemical
18	I-Simple_chemical
)	I-Simple_chemical
F	I-Simple_chemical
]	I-Simple_chemical
deoxyglucose	I-Simple_chemical
(	O
FdG	B-Simple_chemical
)	O
in	O
patients	B-Organism
imaged	O
by	O
positron	O
emission	O
tomography	O
(	O
PET	O
)	O
.	O

[	B-Simple_chemical
(	I-Simple_chemical
18	I-Simple_chemical
)	I-Simple_chemical
F	I-Simple_chemical
]	I-Simple_chemical
deoxyglucose	I-Simple_chemical
uptake	O
correlates	O
with	O
glucose	B-Gene_or_gene_product
transporter	I-Gene_or_gene_product
(	O
GLUT	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
expression	O
,	O
which	O
can	O
be	O
regulated	O
by	O
hypoxia	B-Gene_or_gene_product
-	I-Gene_or_gene_product
inducible	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
1	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
(	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
)	O
.	O

We	O
have	O
previously	O
reported	O
in	O
established	O
breast	B-Cell
lines	I-Cell
that	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
levels	O
in	O
the	O
presence	O
of	O
oxygen	B-Simple_chemical
leads	O
to	O
the	O
Warburg	O
effect	O
.	O

However	O
,	O
glycolysis	O
and	O
GLUT	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
can	O
also	O
be	O
induced	O
independent	O
of	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
by	O
other	O
factors	O
,	O
such	O
as	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Myc	I-Gene_or_gene_product
and	O
phosphorylated	O
Akt	B-Gene_or_gene_product
(	O
pAkt	B-Gene_or_gene_product
)	O
.	O

This	O
study	O
investigates	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
,	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Myc	I-Gene_or_gene_product
,	O
pAkt	B-Gene_or_gene_product
,	O
and	O
aerobic	O
glycolysis	O
in	O
low	O
-	O
passage	O
breast	B-Cell
cancer	I-Cell
cells	I-Cell
under	O
the	O
assumption	O
that	O
these	O
represent	O
the	O
in	O
vivo	O
condition	O
better	O
than	O
established	O
lines	B-Cell
.	O

Similar	O
to	O
in	O
vivo	O
FdG	B-Simple_chemical
-	O
PET	O
or	O
primary	B-Cancer
breast	I-Cancer
cancers	I-Cancer
,	O
rates	O
of	O
glycolysis	O
were	O
diverse	O
,	O
being	O
higher	O
in	O
cells	B-Cell
expressing	O
both	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Myc	I-Gene_or_gene_product
and	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
and	O
lower	O
in	O
cell	B-Cell
lines	I-Cell
low	O
or	O
negative	O
in	O
both	O
transcription	O
factors	O
.	O

No	O
correlations	O
were	O
observed	O
between	O
glycolytic	O
rates	O
and	O
pAkt	B-Gene_or_gene_product
levels	O
.	O

Two	O
of	O
12	O
cell	B-Cell
lines	I-Cell
formed	O
xenografts	B-Cancer
in	O
mice	B-Organism
.	O

Both	O
were	O
positive	O
for	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
and	O
phosphorylated	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Myc	I-Gene_or_gene_product
,	O
and	O
only	O
one	O
was	O
positive	O
for	O
pAkt	B-Gene_or_gene_product
.	O

Glycolysis	O
was	O
affected	O
by	O
pharmacological	O
regulation	O
of	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Myc	I-Gene_or_gene_product
and	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
.	O

These	O
findings	O
suggest	O
that	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Myc	I-Gene_or_gene_product
and	O
/	O
or	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
activities	O
are	O
both	O
involved	O
in	O
the	O
regulation	O
of	O
glycolysis	O
in	O
breast	B-Cancer
cancers	I-Cancer
.	O

Intracerebral	O
grafting	O
of	O
cultured	O
autologous	O
skin	B-Cell
fibroblasts	I-Cell
into	O
the	O
rat	B-Organism
striatum	B-Multi-tissue_structure
:	O
an	O
assessment	O
of	O
graft	B-Tissue
size	O
and	O
ultrastructure	O
.	O

To	O
identify	O
a	O
suitable	O
donor	B-Cell
cell	I-Cell
population	I-Cell
for	O
gene	O
therapy	O
applications	O
to	O
the	O
central	B-Anatomical_system
nervous	I-Anatomical_system
system	I-Anatomical_system
,	O
primary	O
fibroblasts	B-Cell
isolated	O
from	O
skin	B-Multi-tissue_structure
biopsies	I-Multi-tissue_structure
and	O
maintained	O
in	O
culture	B-Cell
are	O
employed	O
as	O
autologous	O
cells	B-Cell
for	O
intracerebral	B-Immaterial_anatomical_entity
grafting	O
within	O
the	O
adult	O
rat	B-Organism
striatum	B-Multi-tissue_structure
.	O

Results	O
from	O
the	O
present	O
investigation	O
reveal	O
that	O
cultured	O
primary	O
skin	B-Cell
fibroblasts	I-Cell
cease	O
to	O
proliferate	O
once	O
they	O
reach	O
confluence	O
;	O
these	O
cells	B-Cell
are	O
thus	O
contact	O
inhibited	O
in	O
vitro	O
.	O

Following	O
implantation	O
within	O
the	O
striatum	B-Multi-tissue_structure
,	O
the	O
volume	O
of	O
the	O
primary	O
fibroblast	B-Tissue
grafts	I-Tissue
,	O
stained	O
immunohistochemically	O
for	O
fibronectin	B-Gene_or_gene_product
,	O
does	O
not	O
differ	O
significantly	O
at	O
3	O
and	O
8	O
weeks	O
.	O

The	O
graft	B-Tissue
size	O
is	O
dependent	O
on	O
the	O
density	O
of	O
the	O
cell	B-Cell
suspension	O
,	O
but	O
not	O
dependent	O
on	O
either	O
the	O
number	O
of	O
passages	O
the	O
cells	B-Cell
are	O
taken	O
through	O
in	O
culture	O
prior	O
to	O
grafting	O
or	O
on	O
the	O
postoperative	O
survival	O
period	O
.	O

Ultrastructural	O
evidence	O
reveals	O
that	O
at	O
8	O
weeks	O
the	O
grafts	B-Tissue
are	O
composed	O
primarily	O
of	O
collagen	B-Gene_or_gene_product
and	O
fibroblasts	B-Cell
with	O
rough	O
endoplasmic	B-Cellular_component
reticulum	I-Cellular_component
and	O
vesicles	B-Cellular_component
.	O

Reactive	O
astrocytic	B-Cellular_component
processes	I-Cellular_component
and	O
phagocytic	B-Cell
cells	I-Cell
are	O
also	O
present	O
in	O
the	O
grafts	B-Tissue
.	O

The	O
grafts	B-Tissue
are	O
extensively	O
vascularized	O
with	O
capillaries	B-Tissue
composed	O
of	O
nonfenestrated	O
endothelium	B-Tissue
;	O
intercellular	B-Cellular_component
junctions	I-Cellular_component
are	O
evident	O
at	O
sites	B-Cellular_component
of	I-Cellular_component
apposition	I-Cellular_component
between	O
endothelial	B-Cell
cells	I-Cell
.	O

It	O
is	O
concluded	O
that	O
primary	O
skin	B-Cell
fibroblasts	I-Cell
are	O
able	O
to	O
survive	O
for	O
at	O
least	O
8	O
weeks	O
following	O
intracerebral	B-Immaterial_anatomical_entity
implantation	O
and	O
continue	O
to	O
synthesize	O
collagen	B-Gene_or_gene_product
and	O
fibronectin	B-Gene_or_gene_product
in	O
vivo	O
.	O

Also	O
,	O
the	O
grafts	B-Tissue
maintain	O
a	O
constant	O
volume	O
between	O
3	O
and	O
8	O
weeks	O
,	O
thereby	O
indicating	O
that	O
primary	O
skin	B-Cell
fibroblasts	I-Cell
do	O
not	O
produce	O
tumors	B-Cancer
.	O

Finally	O
,	O
dynamic	O
host	O
-	O
to	O
-	O
graft	B-Tissue
interactions	O
-	O
-	O
including	O
phagocytic	O
migration	O
,	O
astrocytic	O
hypertrophy	O
and	O
infiltration	O
within	O
the	O
grafts	B-Tissue
,	O
and	O
angiogenesis	O
-	O
-	O
are	O
features	O
that	O
constitute	O
the	O
structural	O
integration	O
of	O
primary	O
skin	B-Cell
fibroblasts	I-Cell
grafted	O
within	O
the	O
adult	O
rat	B-Organism
central	B-Anatomical_system
nervous	I-Anatomical_system
system	I-Anatomical_system
.	O

Angiogenesis	O
associated	O
with	O
visceral	B-Tissue
and	O
subcutaneous	B-Tissue
adipose	I-Tissue
tissue	I-Tissue
in	O
severe	O
human	B-Organism
obesity	O
.	O

OBJECTIVE	O
:	O
The	O
expansion	O
of	O
adipose	B-Tissue
tissue	I-Tissue
is	O
linked	O
to	O
the	O
development	O
of	O
its	O
vasculature	B-Multi-tissue_structure
.	O

However	O
,	O
the	O
regulation	O
of	O
adipose	B-Tissue
tissue	I-Tissue
angiogenesis	O
in	O
humans	B-Organism
has	O
not	O
been	O
extensively	O
studied	O
.	O

Our	O
aim	O
was	O
to	O
compare	O
the	O
angiogenesis	O
associated	O
with	O
subcutaneous	B-Tissue
adipose	I-Tissue
tissue	I-Tissue
(	O
SAT	B-Tissue
)	O
and	O
visceral	B-Tissue
adipose	I-Tissue
tissue	I-Tissue
(	O
VAT	B-Tissue
)	O
from	O
the	O
same	O
obese	O
patients	B-Organism
in	O
an	O
in	O
vivo	O
model	O
.	O

RESEARCH	O
DESIGN	O
AND	O
METHODS	O
:	O
Adipose	B-Tissue
tissue	I-Tissue
samples	I-Tissue
from	O
visceral	B-Tissue
(	O
VAT	B-Tissue
)	O
and	O
subcutaneous	B-Tissue
(	I-Tissue
SAT	I-Tissue
)	I-Tissue
sites	I-Tissue
,	O
obtained	O
from	O
36	O
obese	O
patients	B-Organism
(	O
mean	O
BMI	O
46	O
.	O
5	O
kg	O
/	O
m	O
(	O
2	O
)	O
)	O
during	O
bariatric	O
surgery	O
,	O
were	O
layered	O
on	O
chick	B-Organism
chorioallantoic	B-Multi-tissue_structure
membrane	I-Multi-tissue_structure
(	O
CAM	B-Multi-tissue_structure
)	O
.	O

RESULTS	O
:	O
Both	O
SAT	B-Tissue
and	O
VAT	B-Tissue
expressed	O
angiogenic	O
factors	O
without	O
significant	O
difference	O
for	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
expression	O
.	O

Adipose	B-Tissue
tissue	I-Tissue
layered	O
on	O
CAM	B-Multi-tissue_structure
stimulated	O
angiogenesis	O
.	O

Angiogenic	O
stimulation	O
was	O
macroscopically	O
detectable	O
,	O
with	O
engulfment	O
of	O
the	O
samples	O
,	O
in	O
39	O
%	O
and	O
was	O
evidenced	O
by	O
angiography	O
in	O
59	O
%	O
of	O
the	O
samples	O
.	O

A	O
connection	O
between	O
CAM	B-Multi-tissue_structure
and	O
adipose	B-Tissue
tissue	I-Tissue
vessels	I-Tissue
was	O
evidenced	O
by	O
immunohistochemistry	O
,	O
with	O
recruitment	O
of	O
both	O
avian	B-Organism
and	O
human	B-Organism
endothelial	B-Cell
cells	I-Cell
.	O

The	O
angiogenic	O
potency	O
of	O
adipose	B-Tissue
tissue	I-Tissue
was	O
not	O
related	O
to	O
its	O
localization	O
(	O
with	O
an	O
angiogenic	O
stimulation	O
in	O
60	O
%	O
of	O
SAT	B-Tissue
samples	I-Tissue
and	O
61	O
%	O
of	O
VAT	B-Tissue
samples	I-Tissue
)	O
or	O
to	O
adipocyte	B-Cell
size	O
or	O
inflammatory	O
infiltrate	O
assessed	O
in	O
adipose	B-Tissue
samples	I-Tissue
before	O
the	O
graft	B-Tissue
on	O
CAM	B-Multi-tissue_structure
.	O

Stimulation	O
of	O
angiogenesis	O
by	O
adipose	B-Tissue
tissue	I-Tissue
was	O
nearly	O
abolished	O
by	O
bevacizumab	B-Simple_chemical
,	O
which	O
specifically	O
targets	O
human	B-Organism
VEGF	B-Gene_or_gene_product
.	O

CONCLUSIONS	O
:	O
We	O
have	O
established	O
a	O
model	O
to	O
study	O
the	O
regulation	O
of	O
angiogenesis	O
by	O
human	B-Organism
adipose	B-Tissue
tissue	I-Tissue
.	O

This	O
model	O
highlighted	O
the	O
role	O
of	O
VEGF	B-Gene_or_gene_product
in	O
angiogenesis	O
in	O
both	O
SAT	B-Tissue
and	O
VAT	B-Tissue
.	O

M	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
signals	O
through	O
the	O
MAPK	B-Gene_or_gene_product
/	O
ERK	B-Gene_or_gene_product
pathway	O
via	O
Sp1	B-Gene_or_gene_product
to	O
induce	O
VEGF	B-Gene_or_gene_product
production	O
and	O
induces	O
angiogenesis	O
in	O
vivo	O
.	O

BACKGROUND	O
:	O
M	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
recruits	O
mononuclear	B-Cell
phagocytes	I-Cell
which	O
regulate	O
processes	O
such	O
as	O
angiogenesis	O
and	O
metastases	O
in	O
tumors	B-Cancer
.	O

VEGF	B-Gene_or_gene_product
is	O
a	O
potent	O
activator	O
of	O
angiogenesis	O
as	O
it	O
promotes	O
endothelial	B-Cell
cell	I-Cell
proliferation	O
and	O
new	O
blood	B-Multi-tissue_structure
vessel	I-Multi-tissue_structure
formation	O
.	O

Previously	O
,	O
we	O
reported	O
that	O
in	O
vitro	O
M	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
induces	O
the	O
expression	O
of	O
biologically	O
-	O
active	O
VEGF	B-Gene_or_gene_product
from	O
human	B-Organism
monocytes	B-Cell
.	O

METHODOLOGY	O
AND	O
RESULTS	O
:	O
In	O
this	O
study	O
,	O
we	O
demonstrate	O
the	O
molecular	O
mechanism	O
of	O
M	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
-	O
induced	O
VEGF	B-Gene_or_gene_product
production	O
.	O

Using	O
a	O
construct	O
containing	O
the	O
VEGF	B-Gene_or_gene_product
promoter	O
linked	O
to	O
a	O
luciferase	B-Gene_or_gene_product
reporter	O
,	O
we	O
found	O
that	O
a	O
mutation	O
reducing	O
HIF	B-Gene_or_gene_product
binding	O
to	O
the	O
VEGF	B-Gene_or_gene_product
promoter	O
had	O
no	O
significant	O
effect	O
on	O
luciferase	B-Gene_or_gene_product
production	O
induced	O
by	O
M	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
stimulation	O
.	O

Further	O
analysis	O
revealed	O
that	O
M	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
induced	O
VEGF	B-Gene_or_gene_product
through	O
the	O
MAPK	B-Gene_or_gene_product
/	O
ERK	B-Gene_or_gene_product
signaling	O
pathway	O
via	O
the	O
transcription	O
factor	O
,	O
Sp1	B-Gene_or_gene_product
.	O

Thus	O
,	O
inhibition	O
of	O
either	O
ERK	B-Gene_or_gene_product
or	O
Sp1	B-Gene_or_gene_product
suppressed	O
M	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
-	O
induced	O
VEGF	B-Gene_or_gene_product
at	O
the	O
mRNA	O
and	O
protein	O
level	O
.	O

M	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
also	O
induced	O
the	O
nuclear	B-Cellular_component
localization	O
of	O
Sp1	B-Gene_or_gene_product
,	O
which	O
was	O
blocked	O
by	O
ERK	B-Gene_or_gene_product
inhibition	O
.	O

Finally	O
,	O
mutating	O
the	O
Sp1	B-Gene_or_gene_product
binding	O
sites	O
within	O
the	O
VEGF	B-Gene_or_gene_product
promoter	O
or	O
inhibiting	O
ERK	B-Gene_or_gene_product
decreased	O
VEGF	B-Gene_or_gene_product
promoter	O
activity	O
in	O
M	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
-	O
treated	O
human	B-Organism
monocytes	B-Cell
.	O

To	O
evaluate	O
the	O
biological	O
significance	O
of	O
M	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
induced	O
VEGF	B-Gene_or_gene_product
production	O
,	O
we	O
used	O
an	O
in	O
vivo	O
angiogenesis	O
model	O
to	O
illustrate	O
the	O
ability	O
of	O
M	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
to	O
recruit	O
mononuclear	B-Cell
phagocytes	I-Cell
,	O
increase	O
VEGF	B-Gene_or_gene_product
levels	O
,	O
and	O
enhance	O
angiogenesis	O
.	O

Importantly	O
,	O
the	O
addition	O
of	O
a	O
neutralizing	O
VEGF	B-Gene_or_gene_product
antibody	I-Gene_or_gene_product
abolished	O
M	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
-	O
induced	O
blood	B-Multi-tissue_structure
vessel	I-Multi-tissue_structure
formation	O
.	O

CONCLUSION	O
:	O
These	O
data	O
delineate	O
an	O
ERK	B-Gene_or_gene_product
-	O
and	O
Sp1	B-Gene_or_gene_product
-	O
dependent	O
mechanism	O
of	O
M	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
induced	O
VEGF	B-Gene_or_gene_product
production	O
and	O
demonstrate	O
for	O
the	O
first	O
time	O
the	O
ability	O
of	O
M	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
to	O
induce	O
angiogenesis	O
via	O
VEGF	B-Gene_or_gene_product
in	O
vivo	O
.	O

PI3K	B-Gene_or_gene_product
/	O
Akt	B-Gene_or_gene_product
-	O
dependent	O
transcriptional	O
regulation	O
and	O
activation	O
of	O
BMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
-	O
Smad	B-Gene_or_gene_product
signaling	O
by	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
in	O
metastatic	O
prostate	B-Cell
cancer	I-Cell
cells	I-Cell
.	O

BACKGROUND	O
:	O
Bone	B-Organ
morphogenetic	O
proteins	O
(	O
BMPs	O
)	O
exert	O
osteoinductive	O
effects	O
in	O
prostate	B-Cancer
cancer	I-Cancer
(	O
PC	B-Cancer
)	O
via	O
uncharacterized	O
mechanisms	O
.	O

In	O
this	O
study	O
,	O
we	O
investigated	O
whether	O
the	O
nuclear	B-Cellular_component
transcription	O
factor	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
,	O
implicated	O
in	O
PC	B-Cancer
metastasis	O
,	O
is	O
involved	O
in	O
transcriptional	O
regulation	O
and	O
activation	O
of	O
BMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
or	O
BMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
4	I-Gene_or_gene_product
/	O
Smad	B-Gene_or_gene_product
signaling	O
in	O
PC	B-Cell
cells	I-Cell
.	O

METHODS	O
:	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
inhibition	O
was	O
achieved	O
by	O
IkappaBalpha	B-Gene_or_gene_product
super	O
-	O
repressor	O
adenoviral	B-Organism
vector	O
and	O
activation	O
was	O
monitored	O
by	O
EMSA	O
and	O
reporter	O
assays	O
.	O

BMP	O
expression	O
and	O
activation	O
was	O
measured	O
by	O
PCR	O
and	O
reporter	O
assays	O
.	O

Promoter	O
binding	O
assay	O
was	O
performed	O
by	O
chromatin	B-Cellular_component
immunoprecipitation	O
(	O
ChIP	O
)	O
assay	O
.	O

Smad1	B-Gene_or_gene_product
/	O
5	B-Gene_or_gene_product
/	O
8	B-Gene_or_gene_product
phosphorylation	O
was	O
measured	O
by	O
Western	O
blot	O
analysis	O
.	O

RESULTS	O
:	O
PCR	O
and	O
chimeric	O
BMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
BMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
4	I-Gene_or_gene_product
luciferase	O
assays	O
demonstrate	O
that	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
confers	O
robust	O
and	O
selective	O
activation	O
of	O
BMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
in	O
p65	B-Gene_or_gene_product
overexpressing	O
or	O
rhTNF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
-	O
stimulated	O
PC	B-Cell
cells	I-Cell
.	O

Inhibition	O
of	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
significantly	O
reduced	O
transcript	O
levels	O
and	O
autocrine	O
production	O
of	O
BMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
by	O
rhTNF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
stimulated	O
C4	B-Cell
-	I-Cell
2B	I-Cell
cells	I-Cell
and	O
to	O
a	O
lesser	O
extent	O
by	O
the	O
parental	O
LNCaP	B-Cell
cells	I-Cell
.	O

Selective	O
inhibition	O
of	O
PI3K	B-Gene_or_gene_product
/	O
Akt	B-Gene_or_gene_product
suppressed	O
the	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
-	O
induced	O
BMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
promoter	O
activity	O
.	O

Furthermore	O
,	O
suppression	O
of	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
activation	O
decreased	O
the	O
transcript	O
levels	O
and	O
BMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
-	O
induced	O
phosphorylation	O
of	O
Smad1	B-Gene_or_gene_product
/	O
5	B-Gene_or_gene_product
/	O
8	B-Gene_or_gene_product
,	O
critical	O
downstream	O
targets	O
of	O
BMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
signaling	O
in	O
PC	B-Cell
cells	I-Cell
.	O

Notably	O
,	O
the	O
activation	O
of	O
BMPRII	B-Gene_or_gene_product
by	O
BMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
is	O
required	O
for	O
modulation	O
of	O
Smad	B-Gene_or_gene_product
activation	O
by	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
in	O
PC	B-Cell
cells	I-Cell
.	O

Based	O
on	O
ChIP	O
analysis	O
,	O
the	O
transcriptional	O
regulation	O
of	O
BMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
gene	O
by	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
may	O
be	O
partially	O
attributed	O
to	O
binding	O
to	O
kappab	B-Gene_or_gene_product
site	O
on	O
the	O
BMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
promoter	O
.	O

CONCLUSIONS	O
:	O
The	O
data	O
suggest	O
that	O
PI3K	B-Gene_or_gene_product
/	O
Akt	B-Gene_or_gene_product
-	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
axis	O
may	O
promote	O
PC	B-Multi-tissue_structure
bone	I-Multi-tissue_structure
metastasis	O
in	O
part	O
by	O
regulating	O
transcription	O
and	O
activation	O
of	O
the	O
BMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
-	O
Smad	B-Gene_or_gene_product
signaling	O
cascade	O
in	O
osteotropic	B-Cell
PC	I-Cell
cells	I-Cell
.	O

A	O
novel	O
integrin	B-Gene_or_gene_product
alpha5beta1	I-Gene_or_gene_product
antagonistic	O
peptide	O
,	O
A5	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
screened	O
by	O
Protein	O
Chip	O
system	O
as	O
a	O
potent	O
angiogenesis	O
inhibitor	O
.	O

Integrin	B-Gene_or_gene_product
alpha5beta1	I-Gene_or_gene_product
immobilized	O
on	O
a	O
ProteoChip	O
was	O
used	O
to	O
screen	O
new	O
antagonistic	O
peptides	O
from	O
multiple	O
hexapeptide	O
sub	O
-	O
libraries	O
of	O
the	O
positional	O
scanning	O
synthetic	O
peptide	O
combinatorial	O
library	O
(	O
PS	O
-	O
SPCL	O
)	O
.	O

The	O
integrin	B-Gene_or_gene_product
alpha5beta1	I-Gene_or_gene_product
-	O
Fibronectin	B-Gene_or_gene_product
interaction	O
was	O
demonstrated	O
on	O
the	O
chip	O
.	O

A	O
novel	O
peptide	O
ligand	O
,	O
A5	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
VILVLF	B-Gene_or_gene_product
)	O
,	O
with	O
high	O
affinity	O
to	O
integrin	B-Gene_or_gene_product
alpha5beta1	I-Gene_or_gene_product
was	O
identified	O
from	O
the	O
hexapeptide	O
libraries	O
with	O
this	O
chip	O
-	O
based	O
screening	O
method	O
on	O
the	O
basis	O
of	O
a	O
competitive	O
inhibition	O
assay	O
.	O

A5	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
inhibits	O
the	O
integrin	B-Gene_or_gene_product
-	O
fibronectin	B-Gene_or_gene_product
interaction	O
in	O
a	O
dose	O
-	O
dependent	O
manner	O
(	O
IC	O
(	O
50	O
)	O
;	O
1	O
.	O
56	O
+	O
/	O
-	O
0	O
.	O
28	O
microM	O
.	O

In	O
addition	O
,	O
it	O
inhibits	O
human	B-Cell
umbilical	I-Cell
vein	I-Cell
endothelial	I-Cell
cell	I-Cell
proliferation	O
,	O
migration	O
,	O
adhesion	O
,	O
tubular	B-Multi-tissue_structure
network	I-Multi-tissue_structure
formation	O
,	O
and	O
bFGF	B-Gene_or_gene_product
-	O
induced	O
neovascularization	O
in	O
a	O
chick	B-Organism
chorioallantoic	B-Multi-tissue_structure
membrane	I-Multi-tissue_structure
.	O

These	O
results	O
suggest	O
that	O
A5	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
will	O
be	O
a	O
potent	O
inhibitor	O
of	O
neovascularization	O
.	O

Evaluation	O
of	O
electrical	O
stimulation	O
for	O
ischemic	O
wound	B-Pathological_formation
therapy	O
:	O
a	O
feasibility	O
study	O
using	O
the	O
lapine	B-Organism
wound	B-Pathological_formation
model	O
.	O

Chronic	O
wounds	B-Pathological_formation
are	O
a	O
major	O
secondary	O
complication	O
for	O
many	O
people	B-Organism
with	O
impaired	O
mobility	O
.	O

Electrical	O
stimulation	O
(	O
ES	O
)	O
has	O
been	O
recommended	O
as	O
a	O
adjunctive	O
therapy	O
,	O
however	O
optimal	O
treatment	O
paradigms	O
have	O
not	O
been	O
established	O
.	O

Our	O
group	O
seeks	O
to	O
determine	O
the	O
basic	O
mechanisms	O
underlying	O
ES	O
wound	B-Pathological_formation
therapy	O
,	O
an	O
area	O
where	O
understanding	O
is	O
currently	O
limited	O
.	O

A	O
feasibility	O
study	O
was	O
carried	O
out	O
to	O
develop	O
the	O
Ahn	O
/	O
Mustoe	O
lapine	B-Organism
wound	B-Pathological_formation
model	O
for	O
systematic	O
investigation	O
of	O
the	O
effects	O
of	O
electrical	O
stimulation	O
on	O
ischemic	O
wound	B-Pathological_formation
therapy	O
.	O

A	O
standardized	O
surgical	O
procedure	O
incorporated	O
a	O
hybrid	O
stimulation	O
system	O
comprising	O
an	O
implantable	O
mini	O
-	O
stimulator	O
and	O
surface	O
electrodes	O
,	O
with	O
creation	O
of	O
repeatable	O
ischemic	O
wounds	B-Pathological_formation
.	O

Twenty	O
mature	O
male	O
New	O
Zealand	O
white	O
rabbits	B-Organism
(	O
3	O
kg	O
weight	O
)	O
were	O
employed	O
to	O
evaluate	O
the	O
effects	O
of	O
two	O
empirically	O
selected	O
stimulation	O
paradigms	O
applied	O
continuously	O
for	O
7	O
-	O
21	O
days	O
,	O
using	O
each	O
animal	O
as	O
its	O
own	O
control	O
.	O

Outcomes	O
measures	O
included	O
transcutaneous	O
blood	B-Organism_substance
gas	O
levels	O
,	O
histology	O
,	O
total	O
RNA	O
content	O
and	O
analysis	O
of	O
alpha2	B-Gene_or_gene_product
(	I-Gene_or_gene_product
I	I-Gene_or_gene_product
)	I-Gene_or_gene_product
collagen	I-Gene_or_gene_product
(	O
COL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
I	I-Gene_or_gene_product
)	O
,	O
type	B-Gene_or_gene_product
IV	I-Gene_or_gene_product
collagen	I-Gene_or_gene_product
(	O
COL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
IV	I-Gene_or_gene_product
)	O
,	O
alpha1	B-Gene_or_gene_product
(	I-Gene_or_gene_product
V	I-Gene_or_gene_product
)	I-Gene_or_gene_product
collagen	I-Gene_or_gene_product
(	O
COL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
V	I-Gene_or_gene_product
)	O
,	O
and	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
expression	O
using	O
real	O
-	O
time	O
quantitative	O
PCR	O
.	O

All	O
markers	O
for	O
stimulated	O
wounds	B-Pathological_formation
showed	O
increased	O
activity	O
relative	O
to	O
non	O
-	O
stimulated	O
control	O
wounds	B-Pathological_formation
between	O
7	O
and	O
14	O
days	O
following	O
injury	O
,	O
with	O
peak	O
activity	O
at	O
14	O
days	O
.	O

By	O
21	O
days	O
post	O
-	O
injury	O
,	O
all	O
activity	O
had	O
returned	O
to	O
near	O
baseline	O
level	O
.	O

VEGF	B-Gene_or_gene_product
and	O
COL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
IV	I-Gene_or_gene_product
levels	O
were	O
found	O
to	O
be	O
significantly	O
higher	O
for	O
pattern	O
A	O
(	O
110	O
mus	O
pulse	O
width	O
)	O
compared	O
to	O
pattern	O
B	O
(	O
5	O
mus	O
pulse	O
width	O
)	O
at	O
14	O
days	O
,	O
implying	O
that	O
pattern	O
A	O
may	O
be	O
more	O
effective	O
at	O
promoting	O
angiogenesis	O
.	O

All	O
wounds	B-Pathological_formation
were	O
fully	O
re	O
-	O
epithelialized	O
by	O
10	O
days	O
post	O
-	O
injury	O
.	O

Both	O
COL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
I	I-Gene_or_gene_product
and	O
COL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
V	I-Gene_or_gene_product
showed	O
statistically	O
significant	O
(	O
P	O
less	O
than	O
0	O
.	O
05	O
)	O
increased	O
activity	O
between	O
day	O
7	O
and	O
day	O
14	O
for	O
pattern	O
A	O
,	O
potentially	O
indicating	O
a	O
continued	O
effect	O
on	O
matrix	B-Cellular_component
remodeling	O
.	O

The	O
early	O
closure	O
of	O
all	O
wounds	B-Pathological_formation
implies	O
that	O
the	O
rabbit	B-Organism
ear	B-Organ
model	O
may	O
not	O
be	O
valid	O
for	O
chronic	O
wound	B-Pathological_formation
studies	O
.	O

VEGF	B-Gene_or_gene_product
,	O
angiopoietin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
-	B-Gene_or_gene_product
2	I-Gene_or_gene_product
in	O
bronchial	O
asthma	O
:	O
new	O
molecular	O
targets	O
in	O
airway	B-Multi-tissue_structure
angiogenesis	O
and	O
microvascular	B-Multi-tissue_structure
remodeling	O
.	O

Airway	B-Multi-tissue_structure
angiogenesis	O
and	O
microvascular	B-Multi-tissue_structure
remodeling	O
are	O
known	O
features	O
of	O
bronchial	O
asthma	O
,	O
but	O
the	O
mechanisms	O
of	O
these	O
structural	O
alterations	O
are	O
just	O
beginning	O
to	O
be	O
elucidated	O
.	O

Vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
,	O
one	O
of	O
the	O
most	O
potent	O
angiogenic	O
factors	O
,	O
stimulates	O
endothelial	B-Cell
cell	I-Cell
proliferation	O
and	O
induces	O
the	O
angiogenesis	O
.	O

Recently	O
,	O
considerable	O
attentions	O
have	O
been	O
devoted	O
to	O
the	O
physiological	O
roles	O
of	O
angiopoietin	B-Gene_or_gene_product
(	I-Gene_or_gene_product
Ang	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
-	B-Gene_or_gene_product
2	I-Gene_or_gene_product
as	O
regulatory	O
factors	O
of	O
VEGF	B-Gene_or_gene_product
.	O

Ang	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
has	O
been	O
shown	O
to	O
induce	O
the	O
migration	O
and	O
sprouting	O
of	O
endothelial	B-Cell
cells	I-Cell
,	O
and	O
coexpression	O
of	O
Ang	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
VEGF	B-Gene_or_gene_product
enhanced	O
angiogenesis	O
.	O

In	O
the	O
presence	O
of	O
high	O
levels	O
of	O
VEGF	B-Gene_or_gene_product
,	O
Ang	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
also	O
promotes	O
rapid	O
increase	O
in	O
capillary	B-Tissue
diameter	O
,	O
remodeling	O
of	O
the	O
basal	B-Cellular_component
lamina	I-Cellular_component
,	O
proliferation	O
and	O
migration	O
of	O
endothelial	B-Cell
cells	I-Cell
,	O
and	O
stimulates	O
sprouting	O
of	O
new	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
.	O

Thus	O
,	O
VEGF	B-Gene_or_gene_product
,	O
Ang	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
-	B-Gene_or_gene_product
2	I-Gene_or_gene_product
may	O
play	O
complementary	O
and	O
coordinated	O
roles	O
in	O
airway	B-Multi-tissue_structure
angiogenesis	O
and	O
microvascular	B-Multi-tissue_structure
remodeling	O
,	O
and	O
these	O
structural	O
changes	O
are	O
potentially	O
reversible	O
by	O
therapeutic	O
intervention	O
.	O

The	O
scope	O
of	O
the	O
present	O
review	O
is	O
to	O
discuss	O
from	O
a	O
clinical	O
point	O
of	O
view	O
the	O
potential	O
interactions	O
between	O
VEGF	B-Gene_or_gene_product
and	O
angiopoietins	B-Gene_or_gene_product
in	O
the	O
asthmatic	O
airways	B-Multi-tissue_structure
,	O
and	O
focus	O
on	O
the	O
therapeutic	O
implications	O
targeting	O
for	O
these	O
angiogenic	O
factors	O
.	O

Recently	O
,	O
there	O
is	O
an	O
increasing	O
number	O
of	O
patents	O
which	O
have	O
been	O
focused	O
on	O
the	O
inhibitors	O
of	O
VEGF	B-Gene_or_gene_product
action	O
.	O

These	O
inhibitors	O
are	O
directed	O
towards	O
the	O
receptors	O
of	O
VEGF	B-Gene_or_gene_product
or	O
intracellular	B-Immaterial_anatomical_entity
substrates	O
for	O
the	O
receptors	O
.	O

We	O
will	O
also	O
discuss	O
several	O
patents	O
regarding	O
inhibitors	O
of	O
VEGF	B-Gene_or_gene_product
action	O
in	O
the	O
present	O
review	O
.	O

Effect	O
of	O
transplanted	O
mesenchymal	B-Cell
stem	I-Cell
cells	I-Cell
from	O
rats	B-Organism
of	O
different	O
ages	O
on	O
the	O
improvement	O
of	O
heart	B-Organ
function	O
after	O
acute	O
myocardial	B-Multi-tissue_structure
infarction	O
.	O

BACKGROUND	O
:	O
Mesenchymal	B-Cell
stem	I-Cell
cells	I-Cell
(	O
MSCs	B-Cell
)	O
transplantation	O
is	O
of	O
therapeutic	O
potential	O
after	O
ischemic	O
injury	O
in	O
both	O
experimental	O
and	O
clinical	O
studies	O
.	O

Clinically	O
,	O
elderly	O
patients	B-Organism
are	O
more	O
vulnerable	O
to	O
acute	O
myocardial	B-Multi-tissue_structure
infarction	O
(	O
AMI	O
)	O
.	O

But	O
little	O
is	O
known	O
about	O
the	O
characteristics	O
of	O
young	O
donor	O
-	O
derived	O
MSCs	B-Cell
transplanted	O
to	O
old	O
patients	B-Organism
with	O
AMI	O
.	O

The	O
present	O
study	O
was	O
designed	O
to	O
investigate	O
the	O
effect	O
of	O
transplanted	O
MSCs	B-Cell
from	O
rats	B-Organism
of	O
different	O
ages	O
on	O
the	O
improvement	O
of	O
heart	B-Organ
function	O
after	O
AMI	O
.	O

METHODS	O
:	O
MSCs	B-Cell
from	O
Sprague	B-Organism
-	I-Organism
Dawley	I-Organism
(	I-Organism
SD	I-Organism
)	I-Organism
rats	I-Organism
were	O
isolated	O
and	O
cultured	O
in	O
vitro	O
.	O

The	O
apoptosis	O
characteristics	O
of	O
MSCs	B-Cell
were	O
observed	O
under	O
conditions	O
of	O
ischemia	O
and	O
anoxia	O
.	O

SD	B-Organism
rats	I-Organism
underwent	O
MI	O
received	O
intramyocardial	B-Immaterial_anatomical_entity
injection	O
of	O
MSCs	B-Cell
from	O
young	O
donor	O
rats	B-Organism
(	O
n	O
=	O
8	O
)	O
,	O
old	O
donor	O
rats	B-Organism
(	O
n	O
=	O
8	O
)	O
,	O
respectively	O
.	O

AMI	O
control	O
group	O
received	O
equal	O
volume	O
physiological	O
saline	O
.	O

Immunofluorescence	O
was	O
used	O
to	O
observe	O
the	O
differentiation	O
of	O
the	O
grafted	O
cells	B-Cell
into	O
cardiomyocytes	B-Cell
.	O

Four	O
weeks	O
after	O
cell	B-Cell
transplantation	O
,	O
reverse	O
transcriptase	O
-	O
polymerase	O
chain	O
reaction	O
(	O
RT	O
-	O
PCR	O
)	O
and	O
immunohistochemistry	O
for	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
,	O
VIII	B-Gene_or_gene_product
-	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
immunohistochemistry	O
for	O
vessel	B-Multi-tissue_structure
density	O
,	O
TUNEL	O
,	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
for	O
cardiomyocyte	B-Cell
apoptosis	O
,	O
echocardiography	O
and	O
hemodynamic	O
detection	O
for	O
heart	B-Organ
function	O
were	O
performed	O
.	O

RESULTS	O
:	O
The	O
apoptosis	O
rate	O
of	O
the	O
old	O
donor	O
-	O
derived	O
MSCs	B-Cell
group	O
was	O
significantly	O
higher	O
than	O
that	O
of	O
the	O
young	O
donor	O
-	O
derived	O
MSCs	B-Cell
group	O
under	O
conditions	O
of	O
ischemia	O
and	O
anoxia	O
(	O
P	O
less	O
than	O
0	O
.	O
05	O
)	O
.	O

Engrafted	O
MSCs	B-Cell
survived	O
,	O
proliferated	O
and	O
differentiated	O
into	O
myocardium	B-Cell
-	I-Cell
like	I-Cell
cells	I-Cell
.	O

VEGF	B-Gene_or_gene_product
gene	O
expression	O
and	O
capillary	B-Tissue
density	O
in	O
the	O
old	O
donor	O
-	O
derived	O
group	O
were	O
lower	O
than	O
those	O
in	O
the	O
young	O
donor	O
-	O
derived	O
group	O
but	O
higher	O
than	O
those	O
in	O
the	O
control	O
group	O
(	O
P	O
less	O
than	O
0	O
.	O
05	O
)	O
.	O

The	O
transplantation	O
of	O
old	O
donor	O
-	O
derived	O
MSCs	B-Cell
attenuated	O
apoptosis	O
of	O
cardiomyocytes	B-Cell
in	O
the	O
peri	O
-	O
infarct	O
region	O
compared	O
with	O
the	O
control	O
group	O
and	O
the	O
effect	O
was	O
elevated	O
in	O
young	O
donor	O
-	O
derived	O
MSCs	B-Cell
(	O
P	O
less	O
than	O
0	O
.	O
05	O
)	O
.	O

The	O
heart	B-Organ
functions	O
(	O
left	O
ventricle	B-Multi-tissue_structure
ejection	O
fraction	O
(	O
LVEF	O
)	O
,	O
left	O
ventricle	B-Multi-tissue_structure
fractional	O
shortening	O
(	O
LVFS	O
)	O
)	O
were	O
improved	O
more	O
significantly	O
in	O
the	O
old	O
donor	O
-	O
derived	O
MSCs	B-Cell
group	O
than	O
in	O
the	O
control	O
group	O
and	O
the	O
heart	B-Organ
function	O
in	O
the	O
young	O
donor	O
-	O
derived	O
MSCs	B-Cell
group	O
further	O
improved	O
(	O
P	O
less	O
than	O
0	O
.	O
05	O
)	O
.	O

CONCLUSIONS	O
:	O
Young	O
donor	O
-	O
derived	O
MSCs	B-Cell
can	O
improve	O
heart	B-Organ
function	O
significantly	O
through	O
angiogenesis	O
and	O
decreasing	O
cardiomyocyte	B-Cell
apoptosis	O
when	O
transplanted	O
to	O
the	O
infarcted	O
area	O
.	O

Clinical	O
significance	O
of	O
chicken	B-Gene_or_gene_product
ovalbumin	I-Gene_or_gene_product
upstream	I-Gene_or_gene_product
promoter	I-Gene_or_gene_product
-	I-Gene_or_gene_product
transcription	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
II	I-Gene_or_gene_product
expression	O
in	O
human	B-Organism
colorectal	B-Cancer
cancer	I-Cancer
.	O

Chicken	B-Gene_or_gene_product
ovalbumin	I-Gene_or_gene_product
upstream	I-Gene_or_gene_product
promoter	I-Gene_or_gene_product
-	I-Gene_or_gene_product
transcription	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
II	I-Gene_or_gene_product
(	O
COUP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
TFII	I-Gene_or_gene_product
)	O
plays	O
an	O
essential	O
role	O
in	O
angiogenesis	O
and	O
development	O
.	O

A	O
previous	O
study	O
showed	O
that	O
the	O
expression	O
of	O
COUP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
TFII	I-Gene_or_gene_product
enhanced	O
invasiveness	O
of	O
human	B-Organism
lung	B-Cell
carcinoma	I-Cell
cells	I-Cell
.	O

However	O
,	O
no	O
published	O
data	O
are	O
available	O
concerning	O
the	O
biological	O
and	O
clinical	O
significance	O
of	O
COUP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
TFII	I-Gene_or_gene_product
expression	O
in	O
colorectal	B-Cancer
cancer	I-Cancer
.	O

Thus	O
,	O
our	O
objective	O
was	O
to	O
explore	O
the	O
expression	O
of	O
COUP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
TFII	I-Gene_or_gene_product
in	O
colorectal	B-Cancer
cancer	I-Cancer
as	O
well	O
as	O
its	O
association	O
with	O
clinicopathological	O
features	O
,	O
and	O
to	O
evaluate	O
the	O
role	O
of	O
COUP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
TFII	I-Gene_or_gene_product
as	O
a	O
prognostic	O
indicator	O
in	O
colorectal	B-Cancer
cancer	I-Cancer
.	O

We	O
investigated	O
the	O
presence	O
of	O
COUP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
TFII	I-Gene_or_gene_product
in	O
human	B-Organism
colorectal	B-Tissue
cancer	I-Tissue
tissues	I-Tissue
and	O
adjacent	O
normal	B-Tissue
tissues	I-Tissue
from	O
95	O
primary	O
colorectal	B-Cancer
cancer	I-Cancer
patients	B-Organism
by	O
immunohistochemistry	O
.	O

The	O
correlation	O
between	O
the	O
expression	O
of	O
COUP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
TFII	I-Gene_or_gene_product
and	O
clinicopathologic	O
features	O
was	O
investigated	O
.	O

The	O
3	O
-	O
year	O
disease	O
-	O
free	O
survival	O
(	O
DFS	O
)	O
and	O
overall	O
survival	O
(	O
OS	O
)	O
of	O
patients	B-Organism
with	O
tumors	B-Cancer
expressing	O
different	O
levels	O
of	O
COUP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
TFII	I-Gene_or_gene_product
were	O
evaluated	O
by	O
the	O
Kaplan	O
-	O
Meier	O
method	O
.	O

No	O
significant	O
correlation	O
was	O
found	O
between	O
COUP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
TFII	I-Gene_or_gene_product
expression	O
and	O
age	O
at	O
surgery	O
,	O
gender	O
,	O
histopathologic	O
differentiation	O
,	O
vessel	B-Multi-tissue_structure
invasion	O
,	O
carcinoembryonic	B-Gene_or_gene_product
antigen	I-Gene_or_gene_product
(	O
CEA	B-Gene_or_gene_product
)	O
,	O
or	O
nodal	O
involvement	O
.	O

However	O
,	O
survival	O
analysis	O
showed	O
that	O
the	O
COUP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
TFII	I-Gene_or_gene_product
-	O
positive	O
group	O
had	O
a	O
significantly	O
better	O
OS	O
compared	O
to	O
COUP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
TFII	I-Gene_or_gene_product
-	O
negative	O
group	O
(	O
80	O
.	O
4	O
%	O
vs	O
.	O
57	O
.	O
7	O
%	O
,	O
P	O
=	O
0	O
.	O
0491	O
)	O
.	O

Based	O
on	O
our	O
results	O
,	O
COUP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
TFII	I-Gene_or_gene_product
may	O
represent	O
a	O
biomarker	O
for	O
good	O
prognosis	O
in	O
colorectal	B-Cancer
cancer	I-Cancer
.	O

Homeostatic	O
functions	O
of	O
the	O
p53	B-Gene_or_gene_product
tumor	B-Cancer
suppressor	O
:	O
regulation	O
of	O
energy	O
metabolism	O
and	O
antioxidant	O
defense	O
.	O

The	O
p53	B-Gene_or_gene_product
tumor	B-Cancer
suppressor	O
plays	O
pivotal	O
role	O
in	O
the	O
organism	O
by	O
supervising	O
strict	O
compliance	O
of	O
individual	O
cells	B-Cell
to	O
needs	O
of	O
the	O
whole	O
organisms	O
.	O

It	O
has	O
been	O
widely	O
accepted	O
that	O
p53	B-Gene_or_gene_product
acts	O
in	O
response	O
to	O
stresses	O
and	O
abnormalities	O
in	O
cell	B-Cell
physiology	O
by	O
mobilizing	O
the	O
repair	O
processes	O
or	O
by	O
removing	O
the	O
diseased	O
cells	B-Cell
through	O
initiating	O
the	O
cell	B-Cell
death	O
programs	O
.	O

Recent	O
studies	O
,	O
however	O
,	O
indicate	O
that	O
even	O
under	O
normal	O
physiological	O
conditions	O
certain	O
activities	O
of	O
p53	B-Gene_or_gene_product
participate	O
in	O
homeostatic	O
regulation	O
of	O
metabolic	O
processes	O
and	O
that	O
these	O
activities	O
are	O
important	O
for	O
prevention	O
of	O
cancer	B-Cancer
.	O

These	O
novel	O
functions	O
of	O
p53	B-Gene_or_gene_product
help	O
to	O
align	O
metabolic	O
processes	O
with	O
the	O
proliferation	O
and	O
energy	O
status	O
,	O
to	O
maintain	O
optimal	O
mode	O
of	O
glucose	B-Simple_chemical
metabolism	O
and	O
to	O
boost	O
the	O
energy	O
efficient	O
mitochondrial	B-Cellular_component
respiration	O
in	O
response	O
to	O
ATP	B-Simple_chemical
deficiency	O
.	O

Additional	O
activities	O
of	O
p53	B-Gene_or_gene_product
in	O
non	O
-	O
stressed	O
cells	B-Cell
tune	O
up	O
the	O
antioxidant	O
defense	O
mechanisms	O
reducing	O
the	O
probability	O
of	O
mutations	O
caused	O
by	O
DNA	B-Cellular_component
oxidation	O
under	O
conditions	O
of	O
daily	O
stresses	O
.	O

The	O
deficiency	O
in	O
the	O
p53	B-Gene_or_gene_product
-	O
mediated	O
regulation	O
of	O
glycolysis	O
and	O
mitochondrial	B-Cellular_component
respiration	O
greatly	O
accounts	O
for	O
the	O
deficient	O
respiration	O
of	O
the	O
predominance	O
of	O
aerobic	O
glycolysis	O
in	O
cancer	B-Cell
cells	I-Cell
(	O
the	O
Warburg	O
effect	O
)	O
,	O
while	O
the	O
deficiency	O
in	O
the	O
p53	B-Gene_or_gene_product
-	O
modulated	O
antioxidant	O
defense	O
mechanisms	O
contributes	O
to	O
mutagenesis	O
and	O
additionally	O
boosts	O
the	O
carcinogenesis	O
process	O
.	O

Involvement	O
of	O
PTEN	B-Gene_or_gene_product
promoter	O
methylation	O
in	O
cerebral	B-Pathological_formation
cavernous	I-Pathological_formation
malformations	I-Pathological_formation
.	O

BACKGROUND	O
AND	O
PURPOSE	O
:	O
Cerebral	B-Pathological_formation
cavernous	I-Pathological_formation
malformations	I-Pathological_formation
(	O
CCMs	B-Pathological_formation
)	O
are	O
prevalent	O
cerebral	B-Pathological_formation
vascular	I-Pathological_formation
lesions	I-Pathological_formation
involving	O
aberrant	O
angiogenesis	O
.	O

However	O
,	O
the	O
underlying	O
mechanism	O
is	O
poorly	O
understood	O
.	O

Phosphatase	B-Gene_or_gene_product
and	I-Gene_or_gene_product
tension	I-Gene_or_gene_product
homolog	I-Gene_or_gene_product
deleted	I-Gene_or_gene_product
on	I-Gene_or_gene_product
chromosome	I-Gene_or_gene_product
10	I-Gene_or_gene_product
(	O
PTEN	B-Gene_or_gene_product
)	O
,	O
a	O
tumor	B-Cancer
suppressor	O
,	O
is	O
frequently	O
deficient	O
in	O
various	O
pathologies	O
due	O
to	O
mutation	O
or	O
epigenetic	O
alterations	O
.	O

PTEN	B-Gene_or_gene_product
promoter	O
hypermethylation	O
is	O
a	O
major	O
epigenetic	O
silencing	O
mechanism	O
leading	O
to	O
activation	O
of	O
angiogenesis	O
in	O
tumors	B-Cancer
.	O

The	O
present	O
study	O
aimed	O
to	O
investigate	O
whether	O
PTEN	B-Gene_or_gene_product
promoter	O
methylation	O
was	O
involved	O
in	O
CCMs	B-Pathological_formation
.	O

METHODS	O
:	O
PTEN	B-Gene_or_gene_product
promoter	O
methylation	O
was	O
detected	O
in	O
surgical	O
specimens	O
of	O
CCMs	B-Pathological_formation
(	O
n	O
=	O
69	O
)	O
by	O
methylation	O
-	O
specific	O
polymerase	O
chain	O
reaction	O
.	O

The	O
methylation	O
status	O
was	O
correlated	O
to	O
the	O
clinical	O
manifestations	O
and	O
to	O
PTEN	B-Gene_or_gene_product
expression	O
,	O
which	O
was	O
analyzed	O
by	O
both	O
Western	O
blot	O
and	O
immunohistochemistry	O
.	O

To	O
investigate	O
the	O
endothelial	B-Cell
proliferation	O
and	O
the	O
potential	O
signaling	O
pathways	O
affected	O
by	O
PTEN	B-Gene_or_gene_product
methylation	O
,	O
proliferating	B-Gene_or_gene_product
cell	I-Gene_or_gene_product
nuclear	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
as	O
well	O
as	O
phosphor	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Akt	I-Gene_or_gene_product
and	O
phosphor	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Erk1	I-Gene_or_gene_product
,	I-Gene_or_gene_product
2	I-Gene_or_gene_product
were	O
detected	O
by	O
immunofluorescence	O
and	O
Western	O
blot	O
,	O
respectively	O
,	O
in	O
CCM	B-Pathological_formation
specimens	O
.	O

RESULTS	O
:	O
Methylation	O
-	O
specific	O
polymerase	O
chain	O
reaction	O
revealed	O
PTEN	B-Gene_or_gene_product
promoter	O
methylation	O
in	O
15	O
.	O
9	O
%	O
CCMs	B-Pathological_formation
.	O

Strikingly	O
,	O
5	O
of	O
6	O
familial	O
CCMs	B-Pathological_formation
showed	O
PTEN	B-Gene_or_gene_product
promoter	O
methylation	O
(	O
83	O
.	O
3	O
%	O
)	O
,	O
which	O
was	O
significantly	O
higher	O
than	O
in	O
sporadic	O
cases	O
(	O
9	O
.	O
4	O
%	O
;	O
P	O
<	O
0	O
.	O
001	O
)	O
.	O

In	O
addition	O
,	O
PTEN	B-Gene_or_gene_product
promoter	O
methylation	O
appeared	O
more	O
frequently	O
in	O
multiple	O
CCMs	B-Pathological_formation
,	O
including	O
familial	O
cases	O
(	O
46	O
.	O
7	O
%	O
)	O
,	O
than	O
that	O
in	O
single	O
-	O
lesioned	O
CCMs	B-Pathological_formation
(	O
11	O
.	O
8	O
%	O
;	O
P	O
<	O
0	O
.	O
05	O
)	O
.	O

Immunostaining	O
and	O
Western	O
blot	O
revealed	O
a	O
more	O
significant	O
PTEN	B-Gene_or_gene_product
downregulation	O
in	O
PTEN	B-Gene_or_gene_product
-	O
methylated	O
CCMs	B-Pathological_formation
in	O
comparison	O
to	O
PTEN	B-Gene_or_gene_product
-	O
unmethylated	O
CCMs	B-Pathological_formation
.	O

Reduced	O
PTEN	B-Gene_or_gene_product
expression	O
was	O
inversely	O
correlated	O
to	O
the	O
expression	O
of	O
proliferating	B-Gene_or_gene_product
cell	I-Gene_or_gene_product
nuclear	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
and	O
to	O
the	O
activation	O
of	O
Erk1	B-Gene_or_gene_product
,	O
2	B-Gene_or_gene_product
,	O
but	O
not	O
of	O
Akt	B-Gene_or_gene_product
.	O

CONCLUSIONS	O
:	O
We	O
reported	O
here	O
for	O
the	O
first	O
time	O
the	O
involvement	O
of	O
PTEN	B-Gene_or_gene_product
promoter	O
methylation	O
in	O
CCMs	B-Pathological_formation
,	O
particularly	O
in	O
familial	O
CCMs	B-Pathological_formation
,	O
suggesting	O
this	O
epigenetic	O
alteration	O
as	O
a	O
potential	O
pathomechanism	O
of	O
CCMs	B-Pathological_formation
.	O

The	O
identification	O
of	O
Erk1	B-Gene_or_gene_product
,	O
2	B-Gene_or_gene_product
as	O
triggered	O
signaling	O
in	O
the	O
lesions	B-Pathological_formation
may	O
be	O
valuable	O
for	O
the	O
development	O
of	O
effective	O
therapy	O
for	O
this	O
disease	O
.	O

Aflibercept	B-Simple_chemical
(	O
AVE0005	B-Simple_chemical
)	O
:	O
an	O
alternative	O
strategy	O
for	O
inhibiting	O
tumour	B-Cancer
angiogenesis	O
by	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factors	I-Gene_or_gene_product
.	O

BACKGROUND	O
:	O
Aberrant	O
angiogenesis	O
is	O
a	O
landmark	O
feature	O
in	O
cancer	B-Cancer
,	O
which	O
is	O
important	O
for	O
proliferation	O
,	O
growth	O
and	O
metastasis	O
,	O
and	O
is	O
mediated	O
by	O
various	O
pro	O
-	O
angiogenic	O
factors	O
.	O

The	O
VEGF	B-Gene_or_gene_product
pathway	O
is	O
one	O
of	O
the	O
most	O
important	O
and	O
best	O
-	O
studied	O
angiogenic	O
pathways	O
.	O

Inhibition	O
of	O
this	O
pathway	O
may	O
provide	O
clinical	O
benefits	O
to	O
cancer	B-Cancer
patients	B-Organism
.	O

OBJECTIVES	O
:	O
Strategies	O
to	O
inhibit	O
the	O
VEGF	B-Gene_or_gene_product
pathway	O
,	O
including	O
antibodies	B-Gene_or_gene_product
to	I-Gene_or_gene_product
VEGF	I-Gene_or_gene_product
,	O
antibodies	B-Gene_or_gene_product
to	I-Gene_or_gene_product
the	I-Gene_or_gene_product
extracellular	I-Gene_or_gene_product
domain	I-Gene_or_gene_product
of	I-Gene_or_gene_product
VEGFR	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
or	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
decoy	O
receptors	O
for	O
VEGF	B-Gene_or_gene_product
and	O
tyrosine	B-Simple_chemical
kinase	I-Simple_chemical
inhibitors	I-Simple_chemical
of	O
VEGFRs	B-Gene_or_gene_product
,	O
are	O
summarized	O
.	O

METHODS	O
:	O
This	O
review	O
outlines	O
and	O
compares	O
the	O
latest	O
development	O
of	O
these	O
strategies	O
,	O
with	O
emphasis	O
on	O
aflibercept	B-Simple_chemical
,	O
a	O
novel	O
decoy	O
fusion	O
protein	O
of	O
domain	O
2	O
of	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
domain	O
3	O
of	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
with	O
the	O
Fc	O
fragment	O
of	O
IgG1	B-Gene_or_gene_product
.	O

RESULTS	O
:	O
Aflibercept	B-Simple_chemical
was	O
shown	O
to	O
have	O
early	O
clinical	O
activity	O
.	O

Multiple	O
studies	O
are	O
ongoing	O
to	O
determine	O
the	O
clinical	O
benefits	O
of	O
aflibercept	B-Simple_chemical
in	O
cancer	B-Cancer
patients	B-Organism
.	O

The	O
effect	O
of	O
perfluorocarbon	B-Simple_chemical
-	O
based	O
artificial	O
oxygen	B-Simple_chemical
carriers	O
on	O
tissue	B-Multi-tissue_structure
-	I-Multi-tissue_structure
engineered	I-Multi-tissue_structure
trachea	I-Multi-tissue_structure
.	O

The	O
biological	O
effect	O
of	O
the	O
perfluorocarbon	B-Simple_chemical
-	O
based	O
artificial	O
oxygen	B-Simple_chemical
carrier	O
(	O
Oxygent	B-Simple_chemical
)	O
was	O
investigated	O
in	O
tissue	B-Multi-tissue_structure
-	I-Multi-tissue_structure
engineered	I-Multi-tissue_structure
trachea	I-Multi-tissue_structure
(	O
TET	B-Multi-tissue_structure
)	O
construction	O
.	O

Media	O
supplemented	O
with	O
and	O
without	O
10	O
%	O
Oxygent	B-Simple_chemical
were	O
compared	O
in	O
all	O
assessments	O
.	O

Partial	O
tissue	B-Tissue
oxygen	B-Simple_chemical
tension	O
(	O
PtO	O
(	O
2	O
)	O
)	O
was	O
measured	O
with	O
polarographic	O
microprobes	O
;	O
epithelial	B-Cell
metabolism	O
was	O
monitored	O
by	O
microdialysis	O
inside	O
the	O
TET	B-Tissue
epithelium	I-Tissue
perfused	O
with	O
the	O
medium	O
underneath	O
.	O

Chondrocyte	B-Cell
-	O
DegraPol	O
constructs	O
were	O
cultured	O
for	O
1	O
month	O
with	O
the	O
medium	O
before	O
glycosaminoglycan	B-Simple_chemical
assessment	O
and	O
histology	O
.	O

Tissue	B-Tissue
reaction	O
of	O
TET	B-Tissue
epithelial	I-Tissue
scaffolds	I-Tissue
immersed	O
with	O
the	O
medium	O
was	O
evaluated	O
on	O
the	O
chick	B-Organism
embryo	B-Multi-tissue_structure
chorioallantoic	I-Multi-tissue_structure
membrane	I-Multi-tissue_structure
.	O

Oxygent	B-Simple_chemical
perfusion	O
medium	O
increased	O
the	O
TET	B-Tissue
epithelial	I-Tissue
PtO	O
(	O
2	O
)	O
(	O
51	O
.	O
2	O
+	O
/	O
-	O
0	O
.	O
3	O
mm	O
Hg	O
vs	O
.	O
33	O
.	O
4	O
+	O
/	O
-	O
0	O
.	O
3	O
mm	O
Hg	O
at	O
200	O
microm	O
thickness	O
;	O
12	O
.	O
5	O
+	O
/	O
-	O
0	O
.	O
1	O
mm	O
Hg	O
vs	O
.	O
3	O
.	O
1	O
+	O
/	O
-	O
0	O
.	O
1	O
mm	O
Hg	O
at	O
400	O
microm	O
thickness	O
,	O
p	O
less	O
than	O
0	O
.	O
01	O
)	O
and	O
decreased	O
the	O
lactate	B-Simple_chemical
concentration	O
(	O
0	O
.	O
63	O
+	O
/	O
-	O
0	O
.	O
08	O
vs	O
.	O
0	O
.	O
80	O
+	O
/	O
-	O
0	O
.	O
06	O
mmol	O
/	O
L	O
,	O
p	O
less	O
than	O
0	O
.	O
05	O
)	O
,	O
lactate	B-Simple_chemical
/	O
pyruvate	B-Simple_chemical
(	O
1	O
.	O
87	O
+	O
/	O
-	O
0	O
.	O
26	O
vs	O
.	O
3	O
.	O
36	O
+	O
/	O
-	O
10	O
.	O
13	O
,	O
p	O
less	O
than	O
0	O
.	O
05	O
)	O
,	O
and	O
lactate	B-Simple_chemical
/	O
glucose	B-Simple_chemical
ratios	O
(	O
0	O
.	O
10	O
+	O
/	O
-	O
0	O
.	O
00	O
vs	O
.	O
0	O
.	O
29	O
+	O
/	O
-	O
0	O
.	O
14	O
,	O
p	O
less	O
than	O
0	O
.	O
05	O
)	O
.	O

Chondrocyte	B-Cell
-	O
DegraPol	O
in	O
Oxygent	B-Simple_chemical
group	O
presented	O
lower	O
glycosaminoglycan	B-Simple_chemical
value	O
(	O
0	O
.	O
03	O
+	O
/	O
-	O
0	O
.	O
00	O
vs	O
.	O
0	O
.	O
13	O
+	O
/	O
-	O
0	O
.	O
00	O
,	O
p	O
less	O
than	O
0	O
.	O
05	O
)	O
;	O
histology	O
slides	O
showed	O
poor	O
acid	O
mucopolysaccharides	B-Simple_chemical
formation	O
.	O

Orthogonal	O
polarization	O
spectral	O
imaging	O
showed	O
no	O
difference	O
in	O
functional	O
capillary	B-Tissue
density	O
between	O
the	O
scaffolds	O
cultured	O
on	O
chorioallantoic	B-Multi-tissue_structure
membranes	I-Multi-tissue_structure
.	O

The	O
foreign	O
body	O
reaction	O
was	O
similar	O
in	O
both	O
groups	O
.	O

We	O
conclude	O
that	O
Oxygent	B-Simple_chemical
increases	O
TET	B-Tissue
epithelial	I-Tissue
PtO	O
(	O
2	O
)	O
,	O
improves	O
epithelial	B-Tissue
metabolism	O
,	O
does	O
not	O
impair	O
angiogenesis	O
,	O
and	O
tends	O
to	O
slow	O
cartilage	B-Tissue
tissue	I-Tissue
formation	O
.	O

The	O
semaphorin	B-Gene_or_gene_product
7A	I-Gene_or_gene_product
receptor	O
Plexin	B-Gene_or_gene_product
C1	I-Gene_or_gene_product
is	O
lost	O
during	O
melanoma	B-Cancer
metastasis	O
.	O

The	O
transformation	O
of	O
normal	O
melanocytes	B-Cell
,	O
or	O
melanocyte	B-Cell
stem	I-Cell
cells	I-Cell
,	O
to	O
melanoma	B-Cancer
,	O
is	O
a	O
complex	O
process	O
involving	O
multiple	O
mechanisms	O
.	O

Loss	O
of	O
tumor	B-Cancer
suppressor	O
proteins	O
,	O
which	O
function	O
as	O
brakes	O
on	O
cell	B-Cell
growth	O
,	O
migration	O
,	O
or	O
cell	B-Cell
survival	O
,	O
was	O
recognized	O
early	O
on	O
as	O
an	O
important	O
mechanism	O
for	O
initiation	O
and	O
progression	O
of	O
melanoma	B-Cancer
.	O

Semaphorins	B-Gene_or_gene_product
and	O
their	O
cognate	O
receptors	O
,	O
Plexins	B-Gene_or_gene_product
and	O
neuropilins	B-Gene_or_gene_product
,	O
are	O
involved	O
in	O
neuronal	B-Cell
pathfinding	O
,	O
immune	O
function	O
,	O
and	O
tumor	B-Cancer
progression	O
through	O
effects	O
on	O
blood	B-Multi-tissue_structure
vessel	I-Multi-tissue_structure
growth	O
and	O
cell	B-Cell
migration	O
.	O

Semaphorin	B-Gene_or_gene_product
7A	I-Gene_or_gene_product
(	O
Sema7A	B-Gene_or_gene_product
)	O
is	O
a	O
membrane	B-Cellular_component
-	O
linked	O
semaphorin	B-Gene_or_gene_product
that	O
is	O
expressed	O
by	O
human	B-Organism
keratinocytes	B-Cell
,	O
and	O
we	O
have	O
shown	O
that	O
Sema7A	B-Gene_or_gene_product
binds	O
to	O
human	B-Organism
melanocytes	B-Cell
through	O
beta1	B-Gene_or_gene_product
-	I-Gene_or_gene_product
integrins	I-Gene_or_gene_product
and	O
the	O
Plexin	B-Gene_or_gene_product
C1	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
.	O

Functional	O
studies	O
showed	O
that	O
Sema7A	B-Gene_or_gene_product
stimulates	O
cytoskeletal	B-Cellular_component
reorganization	O
in	O
human	B-Organism
melanocytes	B-Cell
,	O
resulting	O
in	O
adhesion	O
and	O
dendrite	B-Cellular_component
formation	O
.	O

Downstream	O
targets	O
of	O
Plexin	B-Gene_or_gene_product
C1	I-Gene_or_gene_product
signaling	O
in	O
human	B-Organism
melanocytes	B-Cell
include	O
cofilin	B-Gene_or_gene_product
and	O
LIM	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
II	I-Gene_or_gene_product
,	O
both	O
of	O
which	O
are	O
critical	O
mediators	O
of	O
cell	B-Cell
adhesion	O
and	O
migration	O
.	O

In	O
this	O
report	O
,	O
we	O
analyzed	O
the	O
expression	O
of	O
Plexin	B-Gene_or_gene_product
C1	I-Gene_or_gene_product
using	O
immunohistochemistry	O
on	O
sections	O
of	O
primary	O
and	O
matched	O
metastatic	B-Cancer
lesions	I-Cancer
from	O
19	O
subjects	O
and	O
in	O
a	O
large	O
melanoma	B-Cancer
tumor	I-Cancer
microarray	O
.	O

Our	O
data	O
show	O
a	O
significant	O
loss	O
of	O
Plexin	B-Gene_or_gene_product
C1	I-Gene_or_gene_product
in	O
metastatic	B-Cancer
melanoma	I-Cancer
compared	O
with	O
primary	B-Cancer
melanoma	I-Cancer
,	O
suggesting	O
the	O
possibility	O
that	O
the	O
Plexin	B-Gene_or_gene_product
C1	I-Gene_or_gene_product
receptor	O
is	O
a	O
tumor	B-Cancer
suppressor	O
protein	O
for	O
melanoma	B-Cancer
.	O

Coagulation	O
function	O
in	O
patients	B-Organism
with	O
pancreatic	B-Cancer
carcinoma	I-Cancer
.	O

BACKGROUND	O
:	O
The	O
coagulation	O
function	O
in	O
patients	B-Organism
with	O
pancreatic	B-Cancer
carcinoma	I-Cancer
is	O
abnormal	O
and	O
the	O
reason	O
is	O
not	O
very	O
clear	O
.	O

In	O
this	O
study	O
,	O
we	O
retrospectively	O
analyzed	O
the	O
coagulation	O
function	O
in	O
patients	B-Organism
with	O
pancreatic	B-Cancer
carcinoma	I-Cancer
.	O

METHODS	O
:	O
From	O
June	O
2004	O
to	O
December	O
2007	O
,	O
132	O
patients	B-Organism
received	O
diagnosis	O
and	O
treatment	O
in	O
our	O
hospital	O
.	O

The	O
coagulative	O
parameters	O
including	O
the	O
prothrombin	B-Gene_or_gene_product
time	O
,	O
activated	O
partial	O
thromboplastin	B-Gene_or_gene_product
time	O
,	O
and	O
fibrinogen	B-Gene_or_gene_product
levels	O
were	O
collected	O
and	O
studied	O
retrospectively	O
.	O

RESULTS	O
:	O
The	O
average	O
fibrinogen	B-Gene_or_gene_product
levels	O
in	O
patients	B-Organism
with	O
pancreatic	B-Cancer
carcinoma	I-Cancer
,	O
(	O
476	O
.	O
21	O
+	O
/	O
-	O
142	O
.	O
05	O
)	O
mg	O
/	O
dl	O
,	O
were	O
significantly	O
higher	O
than	O
in	O
patients	B-Organism
with	O
cholangiolithiasis	O
,	O
(	O
403	O
.	O
28	O
+	O
/	O
-	O
126	O
.	O
41	O
)	O
mg	O
/	O
dl	O
(	O
P	O
<	O
0	O
.	O
05	O
)	O
.	O

In	O
patients	B-Organism
with	O
pancreatic	B-Cancer
carcinoma	I-Cancer
,	O
the	O
levels	O
of	O
fibrinogen	B-Gene_or_gene_product
in	O
the	O
group	O
with	O
jaundice	O
were	O
significantly	O
higher	O
than	O
in	O
patients	B-Organism
without	O
jaundice	O
(	O
P	O
<	O
0	O
.	O
05	O
)	O
.	O

In	O
patients	B-Organism
who	O
received	O
Pancreaticoduodenectomy	O
,	O
Whipple	O
'	O
s	O
operation	O
,	O
the	O
level	O
of	O
fibrinogen	B-Gene_or_gene_product
in	O
the	O
group	O
with	O
local	O
invasiveness	O
was	O
significantly	O
higher	O
than	O
in	O
the	O
group	O
without	O
invasiveness	O
.	O

The	O
group	O
with	O
lymphatic	B-Organ
metastasis	O
had	O
higher	O
levels	O
than	O
the	O
group	O
without	O
lymphatic	B-Organ
metastasis	O
(	O
P	O
<	O
0	O
.	O
05	O
)	O
.	O

There	O
was	O
no	O
significant	O
difference	O
of	O
intraoperative	O
blood	B-Organism_substance
loss	O
between	O
patients	B-Organism
with	O
vitamin	B-Simple_chemical
K	I-Simple_chemical
,	O
(	O
748	O
.	O
27	O
+	O
/	O
-	O
448	O
.	O
51	O
)	O
ml	O
,	O
and	O
those	O
without	O
vitamin	B-Simple_chemical
K	I-Simple_chemical
,	O
(	O
767	O
.	O
31	O
+	O
/	O
-	O
547	O
.	O
89	O
)	O
ml	O
(	O
P	O
>	O
0	O
.	O
05	O
)	O
.	O

CONCLUSIONS	O
:	O
The	O
level	O
of	O
fibrinogen	B-Gene_or_gene_product
in	O
patients	B-Organism
with	O
pancreatic	B-Cancer
carcinoma	I-Cancer
was	O
elevated	O
.	O

The	O
elevated	O
fibrinogen	B-Gene_or_gene_product
level	O
may	O
be	O
associated	O
with	O
invasiveness	O
and	O
lymphatic	B-Organ
metastasis	O
.	O

Using	O
vitamin	B-Simple_chemical
K	I-Simple_chemical
in	O
perioperation	O
management	O
did	O
not	O
reduce	O
intraoperative	O
blood	B-Organism_substance
loss	O
.	O

Molecular	O
analysis	O
of	O
genetic	O
instability	O
caused	O
by	O
chronic	O
inflammation	O
.	O

Genetic	O
instability	O
is	O
a	O
hallmark	O
of	O
human	B-Organism
cancers	B-Cancer
.	O

It	O
is	O
the	O
driving	O
force	O
for	O
tumor	B-Cancer
development	O
as	O
it	O
facilitates	O
the	O
accumulation	O
of	O
mutations	O
in	O
genes	O
that	O
regulate	O
cell	B-Cell
death	O
and	O
proliferation	O
and	O
therefore	O
promotes	O
malignant	O
transformation	O
.	O

Chronic	O
inflammation	O
is	O
a	O
common	O
underlying	O
condition	O
for	O
human	B-Organism
tumor	B-Cancer
development	O
,	O
accounting	O
for	O
approximately	O
20	O
%	O
of	O
human	B-Organism
cancers	B-Cancer
.	O

TNFalpha	B-Gene_or_gene_product
is	O
an	O
important	O
inflammation	O
cytokine	O
and	O
is	O
crucial	O
to	O
the	O
development	O
of	O
inflammation	O
-	O
associated	O
cancers	B-Cancer
.	O

We	O
have	O
shown	O
that	O
TNFalpha	B-Gene_or_gene_product
can	O
cause	O
DNA	B-Cellular_component
damages	O
through	O
reactive	B-Simple_chemical
oxygen	I-Simple_chemical
species	I-Simple_chemical
(	O
ROS	B-Simple_chemical
)	O
.	O

TNFalpha	B-Gene_or_gene_product
treatment	O
in	O
cultured	O
cells	B-Cell
resulted	O
in	O
increased	O
gene	O
mutations	O
,	O
gene	O
amplification	O
,	O
micronuclei	B-Cellular_component
formation	O
and	O
chromosomal	B-Cellular_component
instability	O
.	O

Antioxidants	O
significantly	O
reduced	O
TNFalpha	B-Gene_or_gene_product
-	O
induced	O
genetic	O
damage	O
.	O

In	O
addition	O
,	O
TNFalpha	B-Gene_or_gene_product
treatment	O
alone	O
led	O
to	O
increased	O
malignant	O
transformation	O
of	O
mouse	B-Organism
embryo	B-Cell
fibroblasts	I-Cell
,	O
which	O
could	O
be	O
partially	O
suppressed	O
by	O
antioxidants	O
.	O

Therefore	O
,	O
genetic	O
instability	O
plays	O
an	O
important	O
role	O
in	O
inflammation	O
-	O
associated	O
cancers	B-Cancer
.	O

PTTG	B-Gene_or_gene_product
overexpression	O
promotes	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
in	O
human	B-Organism
esophageal	B-Cancer
squamous	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
.	O

Human	B-Organism
pituitary	B-Gene_or_gene_product
tumor	I-Gene_or_gene_product
transforming	I-Gene_or_gene_product
gene	I-Gene_or_gene_product
(	O
PTTG	B-Gene_or_gene_product
)	O
overexpression	O
correlates	O
with	O
metastasis	O
in	O
multiple	O
tumors	B-Cancer
,	O
and	O
yet	O
its	O
molecular	O
mechanisms	O
of	O
action	O
remain	O
elusive	O
.	O

We	O
detected	O
PTTG	B-Gene_or_gene_product
overexpression	O
in	O
66	O
%	O
(	O
111	O
of	O
169	O
)	O
of	O
primary	O
esophageal	B-Cancer
squamous	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
(	O
ESCC	B-Cancer
)	O
tumor	B-Tissue
tissues	I-Tissue
by	O
in	O
situ	O
hybridization	O
.	O

PTTG	B-Gene_or_gene_product
overexpression	O
correlated	O
with	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
(	O
P	O
<	O
0	O
.	O
05	O
)	O
.	O

Ectopic	O
PTTG	B-Gene_or_gene_product
overexpression	O
in	O
a	O
representative	O
ESCC	B-Cell
cell	I-Cell
line	I-Cell
,	O
EC9706	B-Cell
,	O
increased	O
in	O
vitro	O
cell	B-Cell
migration	O
and	O
invasion	O
and	O
promoted	O
in	O
vivo	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
.	O

Suppressing	O
PTTG	B-Gene_or_gene_product
expression	O
by	O
siRNA	O
decreased	O
cell	B-Cell
motility	O
in	O
both	O
PTTG	B-Gene_or_gene_product
-	O
HA	O
/	O
EC9706	O
and	O
KYSE150	B-Cell
cells	I-Cell
.	O

By	O
using	O
mass	O
spectrometric	O
analysis	O
,	O
we	O
identified	O
that	O
PTTG	B-Gene_or_gene_product
up	O
-	O
regulated	O
S100A4	B-Gene_or_gene_product
and	O
galectin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
secretion	O
and	O
down	O
-	O
regulated	O
tissue	B-Tissue
inhibitor	O
of	O
metalloproteinase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
secretion	O
to	O
the	O
culture	O
media	O
.	O

PTTG	B-Gene_or_gene_product
induced	O
S100A4	B-Gene_or_gene_product
and	O
galectin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
mRNA	O
and	O
protein	O
expression	O
as	O
assessed	O
by	O
Western	O
blot	O
and	O
reverse	O
transcription	O
-	O
PCR	O
.	O

Attenuating	O
galectin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
expression	O
by	O
siRNA	O
constrained	O
PTTG	B-Gene_or_gene_product
-	O
HA	O
/	O
EC9706	O
cell	O
motility	O
(	O
P	O
<	O
0	O
.	O
05	O
)	O
.	O

PTTG	B-Gene_or_gene_product
activated	O
E	O
-	O
box	O
transcription	O
and	O
induced	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Myc	I-Gene_or_gene_product
protein	O
expression	O
in	O
EC9706	B-Cell
cells	I-Cell
,	O
which	O
in	O
turn	O
may	O
act	O
on	O
an	O
E	O
-	O
box	O
motif	O
within	O
the	O
galectin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
promoter	O
.	O

Chromatin	B-Cellular_component
immunoprecipitation	O
assays	O
further	O
confirmed	O
specific	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Myc	I-Gene_or_gene_product
binding	O
to	O
galectin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
promoter	O
.	O

PTTG	B-Gene_or_gene_product
-	O
induced	O
galectin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
transactivation	O
and	O
expression	O
were	O
mediated	O
by	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Myc	I-Gene_or_gene_product
,	O
and	O
both	O
inductions	O
were	O
suppressed	O
by	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Myc	I-Gene_or_gene_product
RNAi	O
cotranfection	O
.	O

These	O
findings	O
elucidate	O
the	O
molecular	O
mechanisms	O
of	O
PTTG	B-Gene_or_gene_product
overexpression	O
in	O
promoting	O
tumor	B-Cancer
metastasis	O
,	O
whereby	O
up	O
-	O
regulated	O
PTTG	B-Gene_or_gene_product
modulates	O
expression	O
and	O
secretion	O
of	O
metastasis	O
-	O
related	O
factors	O
to	O
facilitate	O
cell	B-Cell
motility	O
.	O

Characterization	O
of	O
the	O
metabolic	O
changes	O
underlying	O
growth	O
factor	O
angiogenic	O
activation	O
:	O
identification	O
of	O
new	O
potential	O
therapeutic	O
targets	O
.	O

Angiogenesis	O
is	O
a	O
fundamental	O
process	O
to	O
normal	O
and	O
abnormal	O
tissue	B-Tissue
growth	O
and	O
repair	O
,	O
which	O
consists	O
of	O
recruiting	O
endothelial	B-Cell
cells	I-Cell
toward	O
an	O
angiogenic	O
stimulus	O
.	O

The	O
cells	B-Cell
subsequently	O
proliferate	O
and	O
differentiate	O
to	O
form	O
endothelial	B-Tissue
tubes	I-Tissue
and	O
capillary	B-Tissue
-	I-Tissue
like	I-Tissue
structures	I-Tissue
.	O

Little	O
is	O
known	O
about	O
the	O
metabolic	O
adaptation	O
of	O
endothelial	B-Cell
cells	I-Cell
through	O
such	O
a	O
transformation	O
.	O

We	O
studied	O
the	O
metabolic	O
changes	O
of	O
endothelial	B-Cell
cell	I-Cell
activation	O
by	O
growth	O
factors	O
using	O
human	B-Cell
umbilical	I-Cell
vein	I-Cell
endothelial	I-Cell
cells	I-Cell
(	O
HUVECs	B-Cell
)	O
,	O
[	B-Simple_chemical
1	I-Simple_chemical
,	I-Simple_chemical
2	I-Simple_chemical
-	I-Simple_chemical
(	I-Simple_chemical
13	I-Simple_chemical
)	I-Simple_chemical
C	I-Simple_chemical
(	I-Simple_chemical
2	I-Simple_chemical
)	I-Simple_chemical
]	I-Simple_chemical
-	I-Simple_chemical
glucose	I-Simple_chemical
and	O
mass	O
isotopomer	O
distribution	O
analysis	O
.	O

The	O
metabolism	O
of	O
[	B-Simple_chemical
1	I-Simple_chemical
,	I-Simple_chemical
2	I-Simple_chemical
-	I-Simple_chemical
(	I-Simple_chemical
13	I-Simple_chemical
)	I-Simple_chemical
C	I-Simple_chemical
(	I-Simple_chemical
2	I-Simple_chemical
)	I-Simple_chemical
]	I-Simple_chemical
-	I-Simple_chemical
glucose	I-Simple_chemical
by	O
HUVEC	B-Cell
allows	O
us	O
to	O
trace	O
many	O
of	O
the	O
main	O
glucose	B-Simple_chemical
metabolic	O
pathways	O
,	O
including	O
glycogen	B-Simple_chemical
synthesis	O
,	O
the	O
pentose	B-Simple_chemical
cycle	O
and	O
the	O
glycolytic	O
pathways	O
.	O

So	O
we	O
established	O
that	O
these	O
pathways	O
were	O
crucial	O
to	O
endothelial	B-Cell
cell	I-Cell
proliferation	O
under	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
and	O
fibroblast	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
FGF	B-Gene_or_gene_product
)	O
stimulation	O
.	O

A	O
specific	O
VEGF	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
inhibitor	O
demonstrated	O
the	O
importance	O
of	O
glycogen	B-Simple_chemical
metabolism	O
and	O
pentose	B-Simple_chemical
cycle	O
pathway	O
.	O

Furthermore	O
,	O
we	O
showed	O
that	O
glycogen	B-Simple_chemical
was	O
depleted	O
in	O
a	O
low	O
glucose	B-Simple_chemical
medium	O
,	O
but	O
conserved	O
under	O
hypoxic	O
conditions	O
.	O

Finally	O
,	O
we	O
demonstrated	O
that	O
direct	O
inhibition	O
of	O
key	O
enzymes	O
to	O
glycogen	B-Simple_chemical
metabolism	O
and	O
pentose	B-Simple_chemical
phosphate	O
pathways	O
reduced	O
HUVEC	B-Cell
viability	O
and	O
migration	O
.	O

In	O
this	O
regard	O
,	O
inhibitors	O
of	O
these	O
pathways	O
have	O
been	O
shown	O
to	O
be	O
effective	O
antitumoral	B-Cancer
agents	O
.	O

To	O
sum	O
up	O
,	O
our	O
data	O
suggest	O
that	O
the	O
inhibition	O
of	O
metabolic	O
pathways	O
offers	O
a	O
novel	O
and	O
powerful	O
therapeutic	O
approach	O
,	O
which	O
simultaneously	O
inhibits	O
tumor	B-Cell
cell	I-Cell
proliferation	O
and	O
tumor	B-Cancer
-	O
induced	O
angiogenesis	O
.	O

Rab	B-Gene_or_gene_product
GTPase	O
regulation	O
of	O
VEGFR2	B-Gene_or_gene_product
trafficking	O
and	O
signaling	O
in	O
endothelial	B-Cell
cells	I-Cell
.	O

OBJECTIVE	O
:	O
Vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
2	I-Gene_or_gene_product
(	O
VEGFR2	B-Gene_or_gene_product
)	O
is	O
a	O
receptor	O
tyrosine	O
kinase	O
that	O
regulates	O
vascular	B-Multi-tissue_structure
physiology	O
.	O

However	O
,	O
mechanism	O
(	O
s	O
)	O
by	O
which	O
VEGFR2	B-Gene_or_gene_product
signaling	O
and	O
trafficking	O
is	O
coordinated	O
are	O
not	O
clear	O
.	O

Here	O
,	O
we	O
have	O
tested	O
endocytic	O
Rab	B-Gene_or_gene_product
GTPases	O
for	O
regulation	O
of	O
VEGFR2	B-Gene_or_gene_product
trafficking	O
and	O
signaling	O
linked	O
to	O
endothelial	B-Cell
cell	I-Cell
migration	O
.	O

METHODS	O
AND	O
RESULTS	O
:	O
Quiescent	O
VEGFR2	B-Gene_or_gene_product
displays	O
endosomal	B-Cellular_component
localization	O
and	O
colocalization	O
with	O
the	O
Rab5a	B-Gene_or_gene_product
GTPase	O
,	O
an	O
early	O
endosome	B-Cellular_component
fusion	O
regulator	O
.	O

Expression	O
of	O
GTP	B-Simple_chemical
or	O
GDP	B-Simple_chemical
-	O
bound	O
Rab5a	B-Gene_or_gene_product
mutants	O
block	O
activated	O
VEGFR2	B-Gene_or_gene_product
trafficking	O
and	O
degradation	O
.	O

Manipulation	O
of	O
Rab7a	B-Gene_or_gene_product
GTPase	O
activity	O
associated	O
with	O
late	B-Cellular_component
endosomes	I-Cellular_component
using	O
overexpression	O
of	O
wild	O
-	O
type	O
or	O
mutant	O
proteins	O
blocks	O
activated	O
VEGFR2	B-Gene_or_gene_product
trafficking	O
and	O
degradation	O
.	O

Depletion	O
of	O
Rab7a	B-Gene_or_gene_product
decreased	O
VEGFR2	B-Gene_or_gene_product
Y1175	O
phosphorylation	O
but	O
increased	O
p42	B-Gene_or_gene_product
/	I-Gene_or_gene_product
44	I-Gene_or_gene_product
(	O
pERK1	B-Gene_or_gene_product
/	I-Gene_or_gene_product
2	I-Gene_or_gene_product
)	O
MAPK	B-Gene_or_gene_product
phosphorylation	O
.	O

Endothelial	B-Cell
cell	I-Cell
migration	O
was	O
increased	O
by	O
Rab5a	B-Gene_or_gene_product
depletion	O
but	O
decreased	O
by	O
Rab7a	B-Gene_or_gene_product
depletion	O
.	O

CONCLUSIONS	O
:	O
Rab5a	B-Gene_or_gene_product
and	O
Rab7a	B-Gene_or_gene_product
regulate	O
VEGFR2	B-Gene_or_gene_product
trafficking	O
toward	O
early	B-Cellular_component
and	O
late	B-Cellular_component
endosomes	I-Cellular_component
.	O

Our	O
data	O
suggest	O
that	O
VEGFR2	B-Gene_or_gene_product
-	O
mediated	O
regulation	O
of	O
endothelial	B-Cell
function	O
is	O
dependent	O
on	O
different	O
but	O
specific	O
Rab	B-Gene_or_gene_product
-	O
mediated	O
GTP	B-Simple_chemical
hydrolysis	O
activity	O
required	O
for	O
endosomal	B-Cellular_component
trafficking	O
.	O

FOXO3a	B-Gene_or_gene_product
elicits	O
a	O
pro	O
-	O
apoptotic	O
transcription	O
program	O
and	O
cellular	B-Cell
response	O
to	O
human	B-Organism
lung	B-Organ
carcinogen	O
nicotine	B-Simple_chemical
-	I-Simple_chemical
derived	I-Simple_chemical
nitrosaminoketone	I-Simple_chemical
(	O
NNK	B-Simple_chemical
)	O
.	O

Long	O
-	O
term	O
carcinogen	O
exposure	O
exerts	O
continuous	O
pressure	O
on	O
key	O
mechanisms	O
that	O
repair	O
or	O
eliminate	O
carcinogen	O
-	O
damaged	O
cells	B-Cell
giving	O
rise	O
to	O
selective	O
failures	O
that	O
contribute	O
to	O
lung	B-Cancer
cancer	I-Cancer
.	O

FOXO3a	B-Gene_or_gene_product
is	O
a	O
transcription	O
factor	O
that	O
elicits	O
a	O
protective	O
response	O
to	O
diverse	O
cellular	B-Cell
stresses	O
.	O

Although	O
implicated	O
as	O
a	O
tumor	B-Cancer
suppressor	O
,	O
its	O
role	O
in	O
sporadic	B-Cancer
cancer	I-Cancer
is	O
uncertain	O
.	O

We	O
recently	O
observed	O
that	O
FOXO3a	B-Gene_or_gene_product
gene	O
inactivation	O
occurs	O
frequently	O
in	O
carcinogen	O
-	O
induced	O
lung	B-Cancer
adenocarcinoma	I-Cancer
(	O
LAC	B-Cancer
)	O
.	O

This	O
suggests	O
that	O
FOXO3a	B-Gene_or_gene_product
may	O
play	O
a	O
role	O
in	O
LAC	B-Cancer
suppression	O
by	O
eliciting	O
a	O
protective	O
response	O
to	O
carcinogenic	O
stress	O
.	O

Here	O
we	O
investigated	O
this	O
possibility	O
by	O
examining	O
the	O
role	O
of	O
FOXO3a	B-Gene_or_gene_product
in	O
the	O
cellular	B-Cell
response	O
to	O
nicotine	B-Simple_chemical
-	I-Simple_chemical
derived	I-Simple_chemical
nitrosaminoketone	I-Simple_chemical
(	O
NNK	B-Simple_chemical
)	O
,	O
a	O
lung	B-Organ
carcinogen	O
implicated	O
as	O
a	O
cause	O
of	O
human	B-Organism
LAC	B-Cancer
.	O

We	O
show	O
that	O
restoration	O
of	O
FOXO3a	B-Gene_or_gene_product
in	O
FOXO3a	B-Gene_or_gene_product
-	O
deficient	O
LAC	B-Cell
cells	I-Cell
increases	O
sensitivity	O
to	O
apoptosis	O
caused	O
by	O
a	O
DNA	B-Cellular_component
-	O
damaging	O
intermediate	O
of	O
NNK	B-Simple_chemical
.	O

Prior	O
to	O
this	O
cellular	B-Cell
outcome	O
,	O
FOXO3a	B-Gene_or_gene_product
is	O
functionally	O
activated	O
and	O
mediates	O
a	O
large	O
-	O
scale	O
transcription	O
program	O
in	O
response	O
to	O
this	O
damage	O
involving	O
a	O
significant	O
modulation	O
of	O
440	O
genes	O
.	O

Genes	O
most	O
significantly	O
represented	O
in	O
this	O
program	O
are	O
those	O
with	O
roles	O
in	O
cell	B-Cell
growth	O
and	O
proliferation	O
>	O
protein	O
synthesis	O
>	O
gene	O
expression	O
>	O
cell	B-Cell
death	O
>	O
cell	B-Cell
cycle	O
.	O

The	O
results	O
of	O
this	O
study	O
show	O
that	O
FOXO3a	B-Gene_or_gene_product
directs	O
an	O
anti	O
-	O
carcinogenic	O
transcription	O
program	O
that	O
culminates	O
in	O
the	O
elimination	O
of	O
carcinogen	O
-	O
damaged	O
cells	B-Cell
.	O

This	O
suggests	O
that	O
FOXO3a	B-Gene_or_gene_product
is	O
a	O
potential	O
suppressor	O
of	O
carcinogenic	O
damage	O
in	O
LAC	B-Cancer
.	O

Bmi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
over	O
-	O
expression	O
in	O
neural	B-Cell
stem	I-Cell
/	I-Cell
progenitor	I-Cell
cells	I-Cell
increases	O
proliferation	O
and	O
neurogenesis	O
in	O
culture	O
but	O
has	O
little	O
effect	O
on	O
these	O
functions	O
in	O
vivo	O
.	O

The	O
polycomb	O
gene	O
Bmi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
is	O
required	O
for	O
the	O
self	O
-	O
renewal	O
of	O
stem	B-Cell
cells	I-Cell
from	O
diverse	O
tissues	B-Tissue
,	O
including	O
the	O
central	B-Anatomical_system
nervous	I-Anatomical_system
system	I-Anatomical_system
(	O
CNS	B-Anatomical_system
)	O
.	O

Bmi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
expression	O
is	O
elevated	O
in	O
most	O
human	B-Organism
gliomas	B-Cancer
,	O
irrespective	O
of	O
grade	O
,	O
raising	O
the	O
question	O
of	O
whether	O
Bmi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
over	O
-	O
expression	O
is	O
sufficient	O
to	O
promote	O
self	O
-	O
renewal	O
or	O
tumorigenesis	O
by	O
CNS	B-Cell
stem	I-Cell
/	I-Cell
progenitor	I-Cell
cells	I-Cell
.	O

To	O
test	O
this	O
we	O
generated	O
Nestin	B-Gene_or_gene_product
-	O
Bmi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
GFP	B-Gene_or_gene_product
transgenic	O
mice	B-Organism
.	O

Analysis	O
of	O
two	O
independent	O
lines	O
with	O
expression	O
in	O
the	O
fetal	B-Anatomical_system
and	O
adult	B-Anatomical_system
CNS	I-Anatomical_system
demonstrated	O
that	O
transgenic	O
neural	B-Cell
stem	I-Cell
cells	I-Cell
formed	O
larger	O
colonies	O
,	O
more	O
self	O
-	O
renewing	O
divisions	O
,	O
and	O
more	O
neurons	B-Cell
in	O
culture	O
.	O

However	O
,	O
in	O
vivo	O
,	O
Bmi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
over	O
-	O
expression	O
had	O
little	O
effect	O
on	O
CNS	B-Cell
stem	I-Cell
cell	I-Cell
frequency	O
,	O
subventricular	B-Immaterial_anatomical_entity
zone	I-Immaterial_anatomical_entity
proliferation	O
,	O
olfactory	B-Multi-tissue_structure
bulb	I-Multi-tissue_structure
neurogenesis	O
,	O
or	O
neurogenesis	O
/	O
gliogenesis	O
during	O
development	O
.	O

Bmi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
transgenic	O
mice	B-Organism
were	O
born	O
with	O
enlarged	O
lateral	B-Multi-tissue_structure
ventricles	I-Multi-tissue_structure
and	O
a	O
minority	O
developed	O
idiopathic	O
hydrocephalus	O
as	O
adults	O
,	O
but	O
none	O
of	O
the	O
transgenic	O
mice	B-Organism
formed	O
detectable	O
CNS	B-Cancer
tumors	I-Cancer
,	O
even	O
when	O
aged	O
.	O

The	O
more	O
pronounced	O
effects	O
of	O
Bmi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
over	O
-	O
expression	O
in	O
culture	O
were	O
largely	O
attributable	O
to	O
the	O
attenuated	O
induction	O
of	O
p16	B-Gene_or_gene_product
(	O
Ink4a	B-Gene_or_gene_product
)	O
and	O
p19	B-Gene_or_gene_product
(	O
Arf	B-Gene_or_gene_product
)	O
in	O
culture	O
,	O
proteins	O
that	O
are	O
generally	O
not	O
expressed	O
by	O
neural	B-Cell
stem	I-Cell
/	I-Cell
progenitor	I-Cell
cells	I-Cell
in	O
young	O
mice	B-Organism
in	O
vivo	O
.	O

Bmi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
over	O
-	O
expression	O
therefore	O
has	O
more	O
pronounced	O
effects	O
in	O
culture	O
and	O
does	O
not	O
appear	O
to	O
be	O
sufficient	O
to	O
induce	O
tumorigenesis	O
in	O
vivo	O
.	O

Erbb2	B-Gene_or_gene_product
suppresses	O
DNA	B-Cellular_component
damage	O
-	O
induced	O
checkpoint	O
activation	O
and	O
UV	O
-	O
induced	O
mouse	B-Organism
skin	B-Organ
tumorigenesis	O
.	O

The	O
Erbb2	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
is	O
activated	O
by	O
UV	O
irradiation	O
,	O
the	O
primary	O
cause	O
of	O
non	B-Cancer
-	I-Cancer
melanoma	I-Cancer
skin	I-Cancer
cancer	I-Cancer
.	O

We	O
hypothesized	O
that	O
Erbb2	B-Gene_or_gene_product
activation	O
contributes	O
to	O
UV	O
-	O
induced	O
skin	B-Organ
tumorigenesis	O
by	O
suppressing	O
cell	B-Cell
cycle	O
arrest	O
.	O

Consistent	O
with	O
this	O
hypothesis	O
,	O
inhibition	O
of	O
Erbb2	B-Gene_or_gene_product
in	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
(	O
Ha	B-Gene_or_gene_product
)	O
transgenic	O
mice	O
before	O
UV	O
exposure	O
resulted	O
in	O
both	O
56	O
%	O
fewer	O
skin	B-Cancer
tumors	I-Cancer
and	O
tumors	B-Cancer
that	O
were	O
70	O
%	O
smaller	O
.	O

Inhibition	O
of	O
the	O
UV	O
-	O
induced	O
activation	O
of	O
Erbb2	B-Gene_or_gene_product
also	O
resulted	O
in	O
milder	O
epidermal	B-Pathological_formation
hyperplasia	I-Pathological_formation
,	O
S	O
-	O
phase	O
accumulation	O
,	O
and	O
decreased	O
levels	O
of	O
the	O
cell	B-Cell
cycle	O
regulator	O
Cdc25a	B-Gene_or_gene_product
,	O
suggesting	O
altered	O
cell	B-Cell
cycle	O
regulation	O
on	O
inhibition	O
of	O
Erbb2	B-Gene_or_gene_product
.	O

Further	O
investigation	O
using	O
inhibition	O
or	O
genetic	O
deletion	O
of	O
Erbb2	B-Gene_or_gene_product
in	O
vitro	O
revealed	O
reduced	O
Cdc25a	B-Gene_or_gene_product
levels	O
and	O
increased	O
S	O
-	O
phase	O
arrest	O
in	O
UV	O
-	O
irradiated	O
cells	B-Cell
lacking	O
Erbb2	B-Gene_or_gene_product
activity	O
.	O

Ectopic	O
expression	O
of	O
Cdc25a	B-Gene_or_gene_product
prevented	O
UV	O
-	O
induced	O
S	O
-	O
phase	O
arrest	O
in	O
keratinocytes	B-Cell
lacking	O
Erbb2	B-Gene_or_gene_product
activity	O
,	O
demonstrating	O
that	O
maintenance	O
of	O
Cdc25a	B-Gene_or_gene_product
by	O
Erbb2	B-Gene_or_gene_product
suppresses	O
cell	B-Cell
cycle	O
arrest	O
.	O

Examination	O
of	O
checkpoint	O
pathway	O
activation	O
upstream	O
of	O
Cdc25a	B-Gene_or_gene_product
revealed	O
Erbb2	B-Gene_or_gene_product
activation	O
did	O
not	O
alter	O
Ataxia	O
Telangiectasia	O
and	O
Rad3	B-Gene_or_gene_product
-	O
related	O
/	O
Ataxia	O
Telangiectasia	O
Mutated	O
activity	O
but	O
increased	O
inhibitory	O
phosphorylation	O
of	O
Chk1	B-Gene_or_gene_product
-	O
Ser	B-Amino_acid
(	I-Amino_acid
280	I-Amino_acid
)	I-Amino_acid
.	O

Since	O
Akt	B-Gene_or_gene_product
phosphorylates	O
Chk1	B-Gene_or_gene_product
-	O
Ser	B-Amino_acid
(	I-Amino_acid
280	I-Amino_acid
)	I-Amino_acid
,	O
the	O
effect	O
of	O
Erbb2	B-Gene_or_gene_product
on	O
phosphatidyl	B-Gene_or_gene_product
inositol	I-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
-	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
(	O
PI3K	B-Gene_or_gene_product
)	O
/	O
Akt	B-Gene_or_gene_product
signaling	O
during	O
UV	O
-	O
induced	O
cell	B-Cell
cycle	O
arrest	O
was	O
determined	O
.	O

Erbb2	B-Gene_or_gene_product
ablation	O
reduced	O
the	O
UV	O
-	O
induced	O
activation	O
of	O
PI3K	B-Gene_or_gene_product
while	O
inhibition	O
of	O
PI3K	B-Gene_or_gene_product
/	O
Akt	B-Gene_or_gene_product
increased	O
UV	O
-	O
induced	O
S	O
-	O
phase	O
arrest	O
.	O

Thus	O
,	O
UV	O
-	O
induced	O
Erbb2	B-Gene_or_gene_product
activation	O
increases	O
skin	B-Organ
tumorigenesis	O
through	O
inhibitory	O
phosphorylation	O
of	O
Chk1	B-Gene_or_gene_product
,	O
Cdc25a	B-Gene_or_gene_product
maintenance	O
,	O
and	O
suppression	O
of	O
S	O
-	O
phase	O
arrest	O
via	O
a	O
PI3K	B-Gene_or_gene_product
/	O
Akt	B-Gene_or_gene_product
-	O
dependent	O
mechanism	O
.	O

The	O
EGFR	B-Gene_or_gene_product
-	O
GEP100	B-Gene_or_gene_product
-	O
Arf6	B-Gene_or_gene_product
-	O
AMAP1	B-Gene_or_gene_product
signaling	O
pathway	O
specific	O
to	O
breast	B-Cancer
cancer	I-Cancer
invasion	O
and	O
metastasis	O
.	O

Tumors	B-Cancer
are	O
tissue	B-Tissue
-	O
specific	O
diseases	O
,	O
and	O
their	O
mechanisms	O
of	O
invasion	O
and	O
metastasis	O
are	O
highly	O
diverse	O
.	O

In	O
breast	B-Cancer
cancer	I-Cancer
,	O
biomarkers	O
that	O
specifically	O
correlate	O
with	O
the	O
invasive	O
phenotypes	O
have	O
not	O
been	O
clearly	O
identified	O
.	O

A	O
small	O
GTPase	O
Arf6	B-Gene_or_gene_product
primarily	O
regulates	O
recycling	O
of	O
plasma	B-Organism_substance
membrane	B-Cellular_component
components	I-Cellular_component
.	O

We	O
have	O
shown	O
that	O
Arf6	B-Gene_or_gene_product
and	O
its	O
effector	O
AMAP1	B-Gene_or_gene_product
(	O
DDEF1	B-Gene_or_gene_product
,	O
DEF1	B-Gene_or_gene_product
,	O
ASAP1	B-Gene_or_gene_product
and	O
centaurin	B-Gene_or_gene_product
beta4	I-Gene_or_gene_product
)	O
are	O
abnormally	O
overexpressed	O
in	O
some	O
breast	B-Cancer
cancers	I-Cancer
and	O
used	O
for	O
their	O
invasion	O
and	O
metastasis	O
.	O

Overexpression	O
of	O
these	O
proteins	O
is	O
independent	O
of	O
the	O
transcriptional	O
upregulation	O
of	O
their	O
genes	O
,	O
and	O
occurs	O
only	O
in	O
highly	O
malignant	B-Cell
breast	I-Cell
cancer	I-Cell
cells	I-Cell
.	O

We	O
recently	O
identified	O
GEP100	B-Gene_or_gene_product
(	O
BRAG2	B-Gene_or_gene_product
)	O
to	O
be	O
responsible	O
for	O
the	O
Arf6	B-Gene_or_gene_product
activation	O
to	O
induce	O
invasion	O
and	O
metastasis	O
,	O
by	O
directly	O
binding	O
to	O
ligand	O
-	O
activated	O
epidermal	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
(	O
EGFR	B-Gene_or_gene_product
)	O
.	O

A	O
series	O
of	O
our	O
studies	O
revealed	O
that	O
for	O
activation	O
of	O
the	O
invasion	O
pathway	O
of	O
EGFR	B-Gene_or_gene_product
,	O
it	O
is	O
prerequisite	O
that	O
Arf6	B-Gene_or_gene_product
and	O
AMAP1	B-Gene_or_gene_product
both	O
are	O
highly	O
overexpressed	O
,	O
and	O
that	O
EGFR	B-Gene_or_gene_product
is	O
activated	O
by	O
ligands	O
.	O

Pathological	O
analyses	O
indicate	O
that	O
a	O
significant	O
large	O
population	O
of	O
human	B-Organism
ductal	B-Cancer
cancers	I-Cancer
may	O
utilize	O
the	O
EGFR	B-Gene_or_gene_product
-	O
GEP100	B-Gene_or_gene_product
-	O
Arf6	B-Gene_or_gene_product
-	O
AMAP1	B-Gene_or_gene_product
pathway	O
for	O
their	O
malignancy	O
.	O

Microenvironments	O
have	O
been	O
highly	O
implicated	O
in	O
the	O
malignancy	O
of	O
mammary	B-Cancer
tumors	I-Cancer
.	O

Our	O
results	O
reveal	O
an	O
aspect	O
of	O
the	O
precise	O
molecular	O
mechanisms	O
of	O
some	O
breast	B-Cancer
cancers	I-Cancer
,	O
in	O
which	O
full	O
invasiveness	O
is	O
not	O
acquired	O
just	O
by	O
intracellular	B-Immaterial_anatomical_entity
alterations	O
of	O
cancer	B-Cell
cells	I-Cell
,	O
but	O
extracellular	B-Immaterial_anatomical_entity
factors	O
from	O
microenvironments	O
may	O
also	O
be	O
necessary	O
.	O

Possible	O
translation	O
of	O
our	O
knowledge	O
to	O
cancer	B-Cancer
therapeutics	O
will	O
also	O
be	O
discussed	O
.	O

Met	B-Gene_or_gene_product
amplification	O
and	O
tumor	B-Cancer
progression	O
in	O
Cdkn2a	B-Gene_or_gene_product
-	O
deficient	O
melanocytes	O
.	O

While	O
many	O
genetic	O
alterations	O
have	O
been	O
identified	O
in	O
melanoma	B-Cancer
,	O
the	O
relevant	O
molecular	O
events	O
that	O
contribute	O
to	O
disease	O
progression	O
are	O
poorly	O
understood	O
.	O

Most	O
primary	O
human	B-Organism
melanomas	B-Cancer
exhibit	O
loss	O
of	O
expression	O
of	O
the	O
CDKN2A	B-Gene_or_gene_product
locus	O
in	O
addition	O
to	O
activation	O
of	O
the	O
canonical	O
mitogen	B-Gene_or_gene_product
-	I-Gene_or_gene_product
activated	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
signaling	O
pathway	O
.	O

In	O
this	O
study	O
,	O
we	O
used	O
a	O
Cdkn2a	B-Gene_or_gene_product
-	O
deficient	O
mouse	B-Organism
melanocyte	B-Cell
cell	I-Cell
line	I-Cell
to	O
screen	O
for	O
secondary	O
genetic	O
events	O
in	O
melanoma	B-Cancer
tumor	I-Cancer
progression	O
.	O

Upon	O
investigation	O
,	O
intrachromosomal	B-Cellular_component
gene	O
amplification	O
of	O
Met	B-Gene_or_gene_product
,	O
a	O
receptor	O
tyrosine	B-Amino_acid
kinase	O
implicated	O
in	O
melanoma	B-Cancer
progression	O
,	O
was	O
identified	O
in	O
Cdkn2a	B-Gene_or_gene_product
-	O
deficient	O
tumors	O
.	O

RNA	O
interference	O
targeting	O
Met	B-Gene_or_gene_product
in	O
these	O
tumor	B-Cell
cells	I-Cell
resulted	O
in	O
a	O
significant	O
delay	O
in	O
tumor	B-Cancer
growth	O
in	O
vivo	O
compared	O
with	O
the	O
control	O
cells	B-Cell
.	O

MET	B-Gene_or_gene_product
expression	O
is	O
rarely	O
detected	O
in	O
primary	O
human	B-Organism
melanoma	B-Cancer
but	O
is	O
frequently	O
observed	O
in	O
metastatic	B-Cancer
disease	I-Cancer
.	O

This	O
study	O
validates	O
a	O
role	O
for	O
Met	B-Gene_or_gene_product
activation	O
in	O
melanoma	B-Cancer
tumor	I-Cancer
progression	O
in	O
the	O
context	O
of	O
Cdkn2a	B-Gene_or_gene_product
deficiency	O
.	O

Activated	O
platelets	B-Cell
enhance	O
ovarian	B-Cell
cancer	I-Cell
cell	I-Cell
invasion	O
in	O
a	O
cellular	B-Cell
model	O
of	O
metastasis	O
.	O

Increased	O
platelet	B-Cell
counts	O
and	O
systemic	O
coagulation	O
activation	O
are	O
associated	O
with	O
ovarian	B-Cancer
cancer	I-Cancer
progression	O
.	O

Platelet	B-Cell
activation	O
occurs	O
in	O
the	O
tumor	B-Cancer
microenvironment	O
and	O
may	O
influence	O
local	O
invasion	O
and	O
metastasis	O
.	O

We	O
used	O
a	O
cellular	B-Cell
model	O
of	O
tumor	B-Cancer
invasion	O
to	O
investigate	O
the	O
effect	O
of	O
activated	O
platelets	B-Cell
on	O
the	O
human	B-Organism
ovarian	B-Cell
cancer	I-Cell
cell	I-Cell
line	I-Cell
,	O
SKOV3	B-Cell
.	O

SKOV3	B-Cell
cells	I-Cell
were	O
exposed	O
to	O
washed	O
,	O
thrombin	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
activating	I-Gene_or_gene_product
peptide	I-Gene_or_gene_product
(	O
TRAP	B-Gene_or_gene_product
)	O
-	O
activated	O
or	O
TRAP	B-Gene_or_gene_product
-	O
naive	O
platelets	B-Cell
under	O
various	O
experimental	O
conditions	O
,	O
and	O
tumor	B-Cell
cell	I-Cell
invasion	O
was	O
assayed	O
in	O
Matrigel	O
chambers	O
.	O

The	O
effect	O
of	O
platelets	B-Cell
on	O
the	O
content	O
of	O
urokinase	B-Gene_or_gene_product
plasminogen	I-Gene_or_gene_product
activator	I-Gene_or_gene_product
(	O
uPA	B-Gene_or_gene_product
)	O
and	O
VEGF	B-Gene_or_gene_product
in	O
SKOV3	B-Cell
cell	I-Cell
conditioned	O
medium	O
was	O
measured	O
using	O
an	O
ELISA	O
assay	O
.	O

TRAP	B-Gene_or_gene_product
-	O
activated	O
platelets	B-Cell
stimulated	O
a	O
dose	O
-	O
dependent	O
increase	O
in	O
SKOV3	B-Cell
cell	I-Cell
invasion	O
.	O

Exposure	O
to	O
activated	O
platelet	B-Cell
membranes	B-Cellular_component
and	O
to	O
soluble	O
proteins	O
contained	O
in	O
activated	O
platelet	B-Cell
releasate	O
both	O
contributed	O
to	O
the	O
observed	O
increase	O
in	O
invasion	O
.	O

The	O
inhibition	O
of	O
platelet	B-Cell
activation	O
with	O
prostaglandin	B-Simple_chemical
E1	I-Simple_chemical
(	O
PGE	B-Simple_chemical
(	I-Simple_chemical
1	I-Simple_chemical
)	I-Simple_chemical
)	O
attenuated	O
the	O
invasive	O
capacity	O
of	O
SKOV3	B-Cell
cells	I-Cell
.	O

Exposure	O
to	O
platelets	B-Cell
resulted	O
in	O
significantly	O
increased	O
uPA	B-Gene_or_gene_product
and	O
VEGF	B-Gene_or_gene_product
content	O
of	O
SKOV3	B-Cell
cell	I-Cell
conditioned	O
medium	O
.	O

Activated	O
platelets	B-Cell
enhance	O
SKOV3	B-Cell
human	B-Organism
ovarian	B-Cell
cancer	I-Cell
cell	I-Cell
invasion	O
through	O
Matrigel	O
and	O
increase	O
the	O
amount	O
of	O
uPA	B-Gene_or_gene_product
and	O
VEGF	B-Gene_or_gene_product
secreted	O
into	O
SKOV3	B-Cell
cell	I-Cell
conditioned	O
medium	O
.	O

If	O
generalizable	O
to	O
additional	O
cell	B-Cell
lines	I-Cell
and	O
human	B-Organism
disease	O
,	O
this	O
observation	O
may	O
partially	O
explain	O
the	O
adverse	O
prognosis	O
associated	O
with	O
thrombocytosis	O
in	O
ovarian	B-Cancer
cancer	I-Cancer
.	O

Platelets	B-Cell
,	O
therefore	O
,	O
may	O
represent	O
a	O
potential	O
target	O
for	O
therapeutic	O
intervention	O
in	O
human	B-Organism
ovarian	B-Cancer
cancer	I-Cancer
.	O

AngiomiRs	B-Gene_or_gene_product
-	O
-	O
key	O
regulators	O
of	O
angiogenesis	O
.	O

The	O
formation	O
of	O
new	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
through	O
the	O
process	O
of	O
angiogenesis	O
is	O
critical	O
in	O
vascular	B-Multi-tissue_structure
development	O
and	O
homeostasis	O
.	O

Aberrant	O
angiogenesis	O
leads	O
to	O
a	O
variety	O
of	O
diseases	O
,	O
such	O
as	O
ischemia	O
and	O
cancer	B-Cancer
.	O

Recent	O
studies	O
have	O
revealed	O
important	O
roles	O
for	O
miRNAs	O
in	O
regulating	O
endothelial	B-Cell
cell	I-Cell
(	O
EC	B-Cell
)	O
function	O
,	O
especially	O
angiogenesis	O
.	O

Mice	B-Organism
with	O
EC	B-Cell
-	O
specific	O
deletion	O
of	O
Dicer	B-Gene_or_gene_product
,	O
a	O
key	O
enzyme	O
for	O
generating	O
miRNAs	O
,	O
display	O
defective	O
postnatal	O
angiogenesis	O
.	O

Specific	O
miRNAs	O
(	O
angiomiRs	B-Gene_or_gene_product
)	O
have	O
recently	O
been	O
shown	O
to	O
regulate	O
angiogenesis	O
in	O
vivo	O
.	O

miRNA	B-Gene_or_gene_product
-	I-Gene_or_gene_product
126	I-Gene_or_gene_product
,	O
an	O
EC	B-Cell
-	O
restricted	O
miRNA	O
,	O
regulates	O
vascular	B-Multi-tissue_structure
integrity	O
and	O
developmental	O
angiogenesis	O
.	O

miR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
378	I-Gene_or_gene_product
,	O
miR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
296	I-Gene_or_gene_product
,	O
and	O
the	O
miR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
17	I-Gene_or_gene_product
-	I-Gene_or_gene_product
92	B-Gene_or_gene_product
cluster	O
contribute	O
to	O
tumor	B-Cancer
angiogenesis	O
.	O

Manipulating	O
angiomiRs	B-Gene_or_gene_product
in	O
the	O
settings	O
of	O
pathological	O
vascularization	O
represents	O
a	O
new	O
therapeutic	O
approach	O
.	O

Promoting	O
angiogenesis	O
via	O
manipulation	O
of	O
VEGF	B-Gene_or_gene_product
responsiveness	O
with	O
notch	B-Gene_or_gene_product
signaling	O
.	O

Promoting	O
angiogenesis	O
via	O
delivery	O
of	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
and	O
other	O
angiogenic	O
factors	O
is	O
both	O
a	O
potential	O
therapy	O
for	O
cardiovascular	B-Anatomical_system
diseases	O
and	O
a	O
critical	O
aspect	O
for	O
tissue	B-Tissue
regeneration	O
.	O

The	O
recent	O
demonstration	O
that	O
VEGF	B-Gene_or_gene_product
signaling	O
is	O
modulated	O
by	O
the	O
Notch	B-Gene_or_gene_product
signaling	O
pathway	O
,	O
however	O
,	O
suggests	O
that	O
inhibiting	O
Notch	B-Gene_or_gene_product
signaling	O
may	O
enhance	O
regional	O
neovascularization	O
,	O
by	O
altering	O
the	O
responsiveness	O
of	O
local	O
endothelial	B-Cell
cells	I-Cell
to	O
angiogenic	O
stimuli	O
.	O

We	O
tested	O
this	O
possibility	O
with	O
in	O
vitro	O
assays	O
using	O
human	B-Organism
endothelial	B-Cell
cells	I-Cell
,	O
as	O
well	O
as	O
in	O
a	O
rodent	O
hindlimb	B-Organism_subdivision
ischemia	O
model	O
.	O

Treatment	O
of	O
cultured	O
human	B-Organism
endothelial	B-Cell
cells	I-Cell
with	O
DAPT	B-Simple_chemical
,	O
a	O
gamma	B-Gene_or_gene_product
secretase	I-Gene_or_gene_product
inhibitor	O
,	O
increased	O
cell	B-Cell
migration	O
and	O
sprout	B-Tissue
formation	O
in	O
response	O
to	O
VEGF	B-Gene_or_gene_product
stimulation	O
with	O
a	O
biphasic	O
dependence	O
on	O
DAPT	B-Simple_chemical
concentration	O
.	O

Further	O
,	O
delivery	O
of	O
an	O
appropriate	O
combination	O
of	O
DAPT	B-Simple_chemical
and	O
VEGF	B-Gene_or_gene_product
from	O
an	O
injectable	O
alginate	O
hydrogel	O
system	O
into	O
ischemic	O
hindlimbs	B-Organism_subdivision
led	O
to	O
a	O
faster	O
recovery	O
of	O
blood	B-Organism_substance
flow	O
than	O
VEGF	B-Gene_or_gene_product
or	O
DAPT	B-Simple_chemical
alone	O
;	O
perfusion	O
levels	O
reached	O
80	O
%	O
of	O
the	O
normal	O
level	O
by	O
week	O
4	O
with	O
combined	O
DAPT	B-Simple_chemical
and	O
VEGF	B-Gene_or_gene_product
delivery	O
.	O

Direct	O
intramuscular	B-Immaterial_anatomical_entity
or	O
intraperitoneal	B-Immaterial_anatomical_entity
injection	O
of	O
DAPT	B-Simple_chemical
did	O
not	O
result	O
in	O
the	O
same	O
level	O
of	O
improvement	O
,	O
suggesting	O
that	O
appropriate	O
presentation	O
of	O
DAPT	B-Simple_chemical
(	O
gel	O
delivery	O
)	O
is	O
important	O
for	O
its	O
activity	O
.	O

DAPT	B-Simple_chemical
delivery	O
from	O
the	O
hydrogels	O
also	O
did	O
not	O
lead	O
to	O
any	O
adverse	O
side	O
effects	O
,	O
in	O
contrast	O
to	O
systemic	O
introduction	O
of	O
DAPT	B-Simple_chemical
.	O

Altogether	O
,	O
these	O
results	O
suggest	O
a	O
new	O
approach	O
to	O
promote	O
angiogenesis	O
by	O
controlling	O
Notch	B-Gene_or_gene_product
signaling	O
,	O
and	O
may	O
provide	O
new	O
options	O
to	O
treat	O
patients	B-Organism
with	O
diseases	O
that	O
diminish	O
angiogenic	O
responsiveness	O
.	O

[	O
Activation	O
of	O
sterol	B-Gene_or_gene_product
regulatory	I-Gene_or_gene_product
element	I-Gene_or_gene_product
binding	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
and	O
its	O
involvement	O
in	O
endothelial	B-Cell
cell	I-Cell
migration	O
]	O

OBJECTIVE	O
:	O
To	O
study	O
the	O
activation	O
of	O
sterol	B-Gene_or_gene_product
regulatory	I-Gene_or_gene_product
element	I-Gene_or_gene_product
binding	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
(	O
SREBP	B-Gene_or_gene_product
)	O
and	O
its	O
critical	O
role	O
in	O
endothelial	B-Cell
cell	I-Cell
migration	O
.	O

METHODS	O
:	O
Bovine	B-Organism
aortic	B-Cell
endothelial	I-Cell
cells	I-Cell
(	O
ECs	B-Cell
)	O
were	O
cultured	O
.	O

The	O
expression	O
of	O
SREBP	B-Gene_or_gene_product
and	O
Cdc42	B-Gene_or_gene_product
were	O
determined	O
by	O
Western	O
blot	O
and	O
quantitative	O
real	O
-	O
time	O
PCR	O
.	O

Moreover	O
,	O
outward	O
growth	O
migration	O
model	O
and	O
transwell	O
chamber	O
assay	O
were	O
used	O
to	O
detect	O
ECs	B-Cell
migration	O
.	O

RESULTS	O
:	O
(	O
1	O
)	O
SREBP	B-Gene_or_gene_product
was	O
activated	O
during	O
ECs	B-Cell
migration	O
.	O

Western	O
blot	O
analysis	O
demonstrated	O
increased	O
active	O
form	O
SREBP	B-Gene_or_gene_product
in	O
migrating	O
as	O
compared	O
to	O
non	O
-	O
migrating	O
ECs	B-Cell
population	O
.	O

SREBP	B-Gene_or_gene_product
activation	O
decreased	O
as	O
ECs	B-Cell
migration	O
slowed	O
;	O
(	O
2	O
)	O
Coincidental	O
with	O
SREBP	B-Gene_or_gene_product
activation	O
,	O
mRNA	O
expression	O
of	O
its	O
target	O
genes	O
such	O
as	O
low	B-Gene_or_gene_product
density	I-Gene_or_gene_product
lipoprotein	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
,	O
HMG	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CoA	I-Gene_or_gene_product
reductase	I-Gene_or_gene_product
,	O
and	O
fatty	B-Gene_or_gene_product
acid	I-Gene_or_gene_product
synthase	I-Gene_or_gene_product
also	O
increased	O
in	O
migrating	O
ECs	B-Cell
population	O
as	O
detected	O
by	O
real	O
-	O
time	O
PCR	O
;	O
(	O
3	O
)	O
Migration	O
induced	O
SREBP	B-Gene_or_gene_product
activation	O
in	O
ECs	B-Cell
was	O
inhibited	O
by	O
SREBP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
acting	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
RNAi	O
and	O
pharmacologically	O
by	O
25	B-Simple_chemical
-	I-Simple_chemical
hydroxycholesterol	I-Simple_chemical
;	O
(	O
4	O
)	O
Inhibition	O
of	O
SREBP	B-Gene_or_gene_product
led	O
to	O
decreased	O
ECs	B-Cell
migration	O
in	O
various	O
models	O
;	O
(	O
5	O
)	O
Cells	B-Cell
genetically	O
deficient	O
in	O
SREBP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
acting	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
,	O
S1P	B-Gene_or_gene_product
,	O
or	O
S2P	B-Gene_or_gene_product
,	O
phenotypically	O
exhibited	O
impaired	O
migration	O
;	O
(	O
6	O
)	O
SREBP	B-Gene_or_gene_product
inhibition	O
in	O
ECs	B-Cell
suppressed	O
the	O
activity	O
of	O
small	O
GTPase	O
Cdc42	B-Gene_or_gene_product
,	O
a	O
key	O
molecule	O
for	O
ECs	B-Cell
motility	O
.	O

CONCLUSIONS	O
:	O
SREBP	B-Gene_or_gene_product
is	O
activated	O
during	O
and	O
plays	O
a	O
critical	O
role	O
in	O
ECs	B-Cell
migration	O
.	O

Targeting	O
SREBP	B-Gene_or_gene_product
could	O
become	O
a	O
novel	O
approach	O
in	O
fighting	O
diseases	O
involving	O
abnormal	O
ECs	B-Cell
migration	O
.	O

Fibroblast	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
2	I-Gene_or_gene_product
-	O
positive	O
fibroblasts	O
provide	O
a	O
suitable	O
microenvironment	O
for	O
tumor	B-Cancer
development	O
and	O
progression	O
in	O
esophageal	B-Cancer
carcinoma	I-Cancer
.	O

PURPOSE	O
:	O
Tumor	B-Cell
fibroblasts	I-Cell
(	O
TF	B-Cell
)	O
have	O
been	O
suggested	O
to	O
play	O
an	O
essential	O
role	O
in	O
the	O
complex	O
process	O
of	O
tumor	B-Cancer
-	O
stroma	B-Tissue
interactions	O
and	O
tumorigenesis	O
.	O

The	O
aim	O
of	O
the	O
present	O
study	O
was	O
to	O
investigate	O
the	O
specific	O
role	O
of	O
TF	B-Cell
in	O
the	O
esophageal	B-Cancer
cancer	I-Cancer
microenvironment	O
.	O

EXPERIMENTAL	O
DESIGN	O
:	O
An	O
Affymetrix	O
expression	O
microarray	O
was	O
used	O
to	O
compare	O
gene	O
expression	O
profiles	O
between	O
six	O
pairs	O
of	O
TFs	B-Cell
and	O
normal	O
fibroblasts	B-Cell
from	O
esophageal	B-Cancer
squamous	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
(	O
ESCC	B-Cancer
)	O
.	O

Differentially	O
expressed	O
genes	O
were	O
identified	O
,	O
and	O
a	O
subset	O
was	O
evaluated	O
by	O
quantitative	O
real	O
-	O
time	O
PCR	O
and	O
immunohistochemistry	O
.	O

RESULTS	O
:	O
About	O
43	O
%	O
(	O
126	O
of	O
292	O
)	O
of	O
known	O
deregulated	O
genes	O
in	O
TFs	B-Cell
were	O
associated	O
with	O
cell	B-Cell
proliferation	O
,	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
remodeling	O
,	O
and	O
immune	O
response	O
.	O

Up	O
-	O
regulation	O
of	O
fibroblast	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
2	I-Gene_or_gene_product
(	O
FGFR2	B-Gene_or_gene_product
)	O
,	O
which	O
showed	O
the	O
most	O
significant	O
change	O
,	O
was	O
detected	O
in	O
all	O
six	O
tested	O
TFs	B-Cell
compared	O
with	O
their	O
paired	O
normal	O
fibroblasts	B-Cell
.	O

A	O
further	O
study	O
found	O
that	O
FGFR2	B-Gene_or_gene_product
-	O
positive	O
fibroblasts	O
were	O
only	O
observed	O
inside	O
the	O
tumor	B-Tissue
tissues	I-Tissue
and	O
not	O
in	O
tumor	B-Tissue
-	I-Tissue
surrounding	I-Tissue
stromal	I-Tissue
tissues	I-Tissue
,	O
suggesting	O
that	O
FGFR2	B-Gene_or_gene_product
could	O
be	O
used	O
as	O
a	O
TF	B-Cell
-	O
specific	O
marker	O
in	O
ESCC	B-Cancer
.	O

Moreover	O
,	O
the	O
conditioned	O
medium	O
from	O
TFs	B-Cell
was	O
found	O
to	O
be	O
able	O
to	O
promote	O
ESCC	B-Cell
tumor	I-Cell
cell	I-Cell
growth	O
,	O
migration	O
,	O
and	O
invasion	O
in	O
vitro	O
.	O

CONCLUSIONS	O
:	O
Our	O
study	O
provides	O
new	O
candidate	O
genes	O
for	O
the	O
esophageal	B-Cancer
cancer	I-Cancer
microenvironment	O
.	O

Based	O
on	O
our	O
results	O
,	O
we	O
hypothesize	O
that	O
FGFR2	B-Gene_or_gene_product
(	O
+	O
)	O
-	O
TFs	O
might	O
provide	O
cancer	B-Cell
cells	I-Cell
with	O
a	O
suitable	O
microenvironment	O
via	O
secretion	O
of	O
proteins	O
that	O
could	O
promote	O
cancer	B-Cancer
development	O
and	O
progression	O
through	O
stimulation	O
of	O
cancer	B-Cell
cell	I-Cell
proliferation	O
,	O
induction	O
of	O
angiogenesis	O
,	O
inhibition	O
of	O
cell	B-Cell
adhesion	O
,	O
enhancement	O
of	O
cell	B-Cell
mobility	O
,	O
and	O
promotion	O
of	O
the	O
epithelial	B-Cell
-	O
mesenchymal	B-Cell
transition	O
.	O

[	O
Role	O
of	O
the	O
tumor	B-Cancer
suppressor	O
ARF	B-Gene_or_gene_product
in	O
oncogenesis	O
]	O

The	O
paper	O
reviews	O
the	O
data	O
available	O
in	O
the	O
literature	O
on	O
a	O
role	O
of	O
the	O
tumor	B-Cancer
suppressor	O
ARF	B-Gene_or_gene_product
in	O
oncogenesis	O
and	O
considers	O
the	O
structure	O
of	O
a	O
gene	O
encoding	O
ARF	B-Gene_or_gene_product
protein	O
.	O

The	O
p53	B-Gene_or_gene_product
-	O
dependent	O
and	O
p	B-Gene_or_gene_product
-	I-Gene_or_gene_product
53	I-Gene_or_gene_product
-	O
independent	O
functions	O
of	O
this	O
protein	O
are	O
under	O
many	O
studies	O
.	O

There	O
is	O
evidence	O
for	O
the	O
implication	O
of	O
ARF	B-Gene_or_gene_product
in	O
angiogenesis	O
.	O

There	O
is	O
more	O
and	O
more	O
information	O
on	O
the	O
role	O
of	O
ARF	B-Gene_or_gene_product
in	O
the	O
regulation	O
of	O
a	O
cell	B-Cell
cycle	O
,	O
apoptosis	O
,	O
and	O
autophagy	O
.	O

The	O
importance	O
of	O
this	O
tumor	B-Cancer
suppressor	O
in	O
the	O
mechanisms	O
of	O
carcinogenesis	O
is	O
beyond	O
question	O
as	O
the	O
inactivation	O
of	O
ARF	B-Gene_or_gene_product
suppressor	O
activity	O
leads	O
to	O
the	O
rapid	O
growth	O
of	O
neoplasia	B-Cancer
.	O

However	O
,	O
the	O
exact	O
mechanisms	O
of	O
ARF	B-Gene_or_gene_product
action	O
yet	O
remain	O
unclear	O
and	O
require	O
further	O
studies	O
by	O
different	O
specialists	O
at	O
both	O
the	O
molecular	O
genetic	O
and	O
other	O
levels	O
of	O
investigation	O
.	O

Mechanical	O
regulation	O
of	O
the	O
proangiogenic	O
factor	O
CCN1	B-Gene_or_gene_product
/	O
CYR61	B-Gene_or_gene_product
gene	O
requires	O
the	O
combined	O
activities	O
of	O
MRTF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
A	I-Gene_or_gene_product
and	O
CREB	B-Gene_or_gene_product
-	I-Gene_or_gene_product
binding	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
histone	I-Gene_or_gene_product
acetyltransferase	I-Gene_or_gene_product
.	O

Smooth	B-Tissue
muscle	I-Tissue
-	I-Tissue
rich	I-Tissue
tissues	I-Tissue
respond	O
to	O
mechanical	O
overload	O
by	O
an	O
adaptive	O
hypertrophic	O
growth	O
combined	O
with	O
activation	O
of	O
angiogenesis	O
,	O
which	O
potentiates	O
their	O
mechanical	O
overload	O
-	O
bearing	O
capabilities	O
.	O

Neovascularization	O
is	O
associated	O
with	O
mechanical	O
strain	O
-	O
dependent	O
induction	O
of	O
angiogenic	O
factors	O
such	O
as	O
CCN1	B-Gene_or_gene_product
,	O
an	O
immediate	O
-	O
early	O
gene	O
-	O
encoded	O
matricellular	O
molecule	O
critical	O
for	O
vascular	B-Multi-tissue_structure
development	O
and	O
repair	O
.	O

Here	O
we	O
have	O
demonstrated	O
that	O
mechanical	O
strain	O
-	O
dependent	O
induction	O
of	O
the	O
CCN1	B-Gene_or_gene_product
gene	O
involves	O
signaling	O
cascades	O
through	O
RhoA	B-Gene_or_gene_product
-	O
mediated	O
actin	B-Cellular_component
remodeling	O
and	O
the	O
p38	B-Gene_or_gene_product
stress	I-Gene_or_gene_product
-	I-Gene_or_gene_product
activated	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
(	O
SAPK	B-Gene_or_gene_product
)	O
.	O

Actin	B-Gene_or_gene_product
signaling	O
controls	O
serum	B-Gene_or_gene_product
response	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
SRF	B-Gene_or_gene_product
)	O
activity	O
via	O
SRF	B-Gene_or_gene_product
interaction	O
with	O
the	O
myocardin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
related	I-Gene_or_gene_product
transcriptional	I-Gene_or_gene_product
activator	I-Gene_or_gene_product
(	I-Gene_or_gene_product
MRTF	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	I-Gene_or_gene_product
A	I-Gene_or_gene_product
and	O
tethering	O
to	O
a	O
single	O
CArG	O
box	O
sequence	O
within	O
the	O
CCN1	B-Gene_or_gene_product
promoter	O
.	O

Such	O
activity	O
was	O
abolished	O
in	O
mechanically	O
stimulated	O
mouse	B-Organism
MRTF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
A	I-Gene_or_gene_product
(	O
-	O
/	O
-	O
)	O
cells	O
or	O
upon	O
inhibition	O
of	O
CREB	B-Gene_or_gene_product
-	I-Gene_or_gene_product
binding	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
(	I-Gene_or_gene_product
CBP	I-Gene_or_gene_product
)	I-Gene_or_gene_product
histone	I-Gene_or_gene_product
acetyltransferase	I-Gene_or_gene_product
(	I-Gene_or_gene_product
HAT	I-Gene_or_gene_product
)	I-Gene_or_gene_product
either	O
pharmacologically	O
or	O
by	O
siRNAs	O
.	O

Mechanical	O
strain	O
induced	O
CBP	B-Gene_or_gene_product
-	O
mediated	O
acetylation	O
of	O
histones	B-Gene_or_gene_product
3	I-Gene_or_gene_product
and	O
4	B-Gene_or_gene_product
at	O
the	O
SRF	B-Gene_or_gene_product
-	O
binding	O
site	O
and	O
within	O
the	O
CCN1	B-Gene_or_gene_product
gene	O
coding	O
region	O
.	O

Inhibition	O
of	O
p38	B-Gene_or_gene_product
SAPK	I-Gene_or_gene_product
reduced	O
CBP	B-Gene_or_gene_product
HAT	I-Gene_or_gene_product
activity	O
and	O
its	O
recruitment	O
to	O
the	O
SRF	B-Gene_or_gene_product
.	O
MRTF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
A	I-Gene_or_gene_product
complex	O
,	O
whereas	O
enforced	O
induction	O
of	O
p38	B-Gene_or_gene_product
by	O
upstream	O
activators	O
(	O
e	O
.	O
g	O
.	O
MKK3	B-Gene_or_gene_product
and	O
MKK6	B-Gene_or_gene_product
)	O
enhanced	O
both	O
CBP	B-Gene_or_gene_product
HAT	I-Gene_or_gene_product
and	O
CCN1	B-Gene_or_gene_product
promoter	O
activities	O
.	O

Similarly	O
,	O
mechanical	O
overload	O
-	O
induced	O
CCN1	B-Gene_or_gene_product
gene	O
expression	O
in	O
vivo	O
was	O
associated	O
with	O
nuclear	O
localization	O
of	O
MRTF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
A	I-Gene_or_gene_product
and	O
enrichment	O
of	O
the	O
CCN1	B-Gene_or_gene_product
promoter	O
with	O
both	O
MRTF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
A	I-Gene_or_gene_product
and	O
acetylated	O
histone	B-Gene_or_gene_product
H3	I-Gene_or_gene_product
.	O

Taken	O
together	O
,	O
these	O
data	O
suggest	O
that	O
signal	O
-	O
controlled	O
activation	O
of	O
SRF	B-Gene_or_gene_product
,	O
MRTF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
A	I-Gene_or_gene_product
,	O
and	O
CBP	B-Gene_or_gene_product
provides	O
a	O
novel	O
connection	O
between	O
mechanical	O
stimuli	O
and	O
angiogenic	O
gene	O
expression	O
.	O

Angiogenesis	O
in	O
platelet	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
cell	I-Gene_or_gene_product
adhesion	I-Gene_or_gene_product
molecule	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
null	O
mice	O
.	O

Platelet	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
cell	I-Gene_or_gene_product
adhesion	I-Gene_or_gene_product
molecule	I-Gene_or_gene_product
(	I-Gene_or_gene_product
PECAM	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
has	O
been	O
previously	O
implicated	O
in	O
endothelial	B-Cell
cell	I-Cell
migration	O
;	O
additionally	O
,	O
anti	B-Gene_or_gene_product
-	I-Gene_or_gene_product
PECAM	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
antibodies	I-Gene_or_gene_product
have	O
been	O
shown	O
to	O
inhibit	O
in	O
vivo	O
angiogenesis	O
.	O

Studies	O
were	O
therefore	O
performed	O
with	O
PECAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
null	O
mice	O
to	O
further	O
define	O
the	O
involvement	O
of	O
PECAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
in	O
blood	B-Multi-tissue_structure
vessel	I-Multi-tissue_structure
formation	O
.	O

Vascularization	O
of	O
subcutaneous	O
Matrigel	O
implants	O
as	O
well	O
as	O
tumor	B-Cancer
angiogenesis	O
were	O
both	O
inhibited	O
in	O
PECAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
null	O
mice	O
.	O

Reciprocal	O
bone	B-Multi-tissue_structure
marrow	I-Multi-tissue_structure
transplants	O
that	O
involved	O
both	O
wild	O
-	O
type	O
and	O
PECAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
deficient	O
mice	O
revealed	O
that	O
the	O
impaired	O
angiogenic	O
response	O
resulted	O
from	O
a	O
loss	O
of	O
endothelial	B-Cell
,	O
but	O
not	O
leukocyte	B-Cell
,	O
PECAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

In	O
vitro	O
wound	B-Pathological_formation
migration	O
and	O
single	O
-	O
cell	B-Cell
motility	O
by	O
PECAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
null	O
endothelial	O
cells	O
were	O
also	O
compromised	O
.	O

In	O
addition	O
,	O
filopodia	B-Cellular_component
formation	O
,	O
a	O
feature	O
of	O
motile	O
cells	B-Cell
,	O
was	O
inhibited	O
in	O
PECAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
null	O
endothelial	O
cells	O
as	O
well	O
as	O
in	O
human	B-Organism
endothelial	B-Cell
cells	I-Cell
treated	O
with	O
either	O
anti	B-Gene_or_gene_product
-	I-Gene_or_gene_product
PECAM	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
antibody	I-Gene_or_gene_product
or	O
PECAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
siRNA	O
.	O

Furthermore	O
,	O
the	O
expression	O
of	O
PECAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
promoted	O
filopodia	B-Cellular_component
formation	O
and	O
increased	O
the	O
protein	O
expression	O
levels	O
of	O
Cdc42	B-Gene_or_gene_product
,	O
a	O
Rho	B-Gene_or_gene_product
GTPase	O
that	O
is	O
known	O
to	O
promote	O
the	O
formation	O
of	O
filopodia	B-Cellular_component
.	O

In	O
the	O
developing	O
retinal	B-Multi-tissue_structure
vasculature	I-Multi-tissue_structure
,	O
numerous	O
,	O
long	B-Cellular_component
filamentous	I-Cellular_component
filopodia	I-Cellular_component
,	O
emanating	O
from	O
endothelial	B-Cell
cells	I-Cell
at	O
the	O
tips	O
of	O
angiogenic	O
sprouts	B-Tissue
,	O
were	O
observed	O
in	O
wild	O
-	O
type	O
animals	O
,	O
but	O
to	O
a	O
lesser	O
extent	O
in	O
the	O
PECAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
null	O
mice	O
.	O

Together	O
,	O
these	O
data	O
further	O
establish	O
the	O
involvement	O
of	O
endothelial	B-Cell
PECAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
in	O
angiogenesis	O
and	O
suggest	O
that	O
,	O
in	O
vivo	O
,	O
PECAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
may	O
stimulate	O
endothelial	B-Cell
cell	I-Cell
motility	O
by	O
promoting	O
the	O
formation	O
of	O
filopodia	B-Cellular_component
.	O

Desipramine	B-Simple_chemical
inhibits	O
the	O
growth	O
of	O
a	O
mouse	B-Organism
skin	B-Cell
squamous	I-Cell
cell	I-Cell
carcinoma	I-Cell
cell	I-Cell
line	I-Cell
and	O
affects	O
glucocorticoid	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
-	O
mediated	O
transcription	O
.	O

The	O
purpose	O
of	O
this	O
study	O
was	O
to	O
examine	O
the	O
effect	O
of	O
tricyclic	O
antidepressant	O
desipramine	B-Simple_chemical
(	O
DMI	B-Simple_chemical
)	O
on	O
the	O
growth	O
inhibition	O
and	O
translocation	O
of	O
the	O
glucocorticoid	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
(	O
GR	B-Gene_or_gene_product
)	O
from	O
the	O
cytoplasm	B-Organism_substance
to	O
the	O
nucleus	B-Cellular_component
in	O
cancerous	B-Cell
and	O
noncancerous	B-Cell
cell	I-Cell
lines	I-Cell
and	O
the	O
effect	O
of	O
DMI	B-Simple_chemical
on	O
GR	B-Gene_or_gene_product
-	O
mediated	O
transcription	O
.	O

Nontumorigenic	O
,	O
immortalized	O
keratinocytes	B-Cell
cell	I-Cell
line	I-Cell
(	O
3PC	B-Cell
)	O
,	O
papilloma	B-Cell
(	O
MT1	B-Cell
/	I-Cell
2	I-Cell
)	O
,	O
and	O
squamous	B-Cell
cell	I-Cell
carcinoma	I-Cell
(	O
Ca3	B-Cell
/	I-Cell
7	I-Cell
)	O
cell	B-Cell
lines	I-Cell
were	O
initially	O
used	O
to	O
study	O
the	O
cell	B-Cell
growth	O
inhibition	O
by	O
DMI	B-Simple_chemical
.	O

Although	O
,	O
the	O
growth	O
of	O
all	O
three	O
cell	B-Cell
lines	I-Cell
was	O
suppressed	O
by	O
DMI	B-Simple_chemical
,	O
it	O
was	O
more	O
effective	O
in	O
Ca3	B-Cell
/	I-Cell
7	I-Cell
cells	I-Cell
.	O

Therefore	O
,	O
we	O
next	O
examined	O
the	O
effect	O
of	O
DMI	B-Simple_chemical
on	O
Ca3	B-Cell
/	I-Cell
7	I-Cell
cells	I-Cell
,	O
resistant	O
to	O
growth	O
inhibition	O
by	O
the	O
synthetic	O
glucocorticoid	B-Simple_chemical
fluocinolone	I-Simple_chemical
acetonide	I-Simple_chemical
(	O
FA	B-Simple_chemical
)	O
.	O

DMI	B-Simple_chemical
inhibited	O
cell	B-Cell
proliferation	O
in	O
a	O
time	O
-	O
dependent	O
manner	O
.	O

The	O
translocation	O
of	O
GR	B-Gene_or_gene_product
was	O
induced	O
by	O
FA	B-Simple_chemical
alone	O
,	O
DMI	B-Simple_chemical
alone	O
,	O
and	O
combination	O
of	O
both	O
agents	O
.	O

FA	B-Simple_chemical
induced	O
GR	B-Gene_or_gene_product
-	O
mediated	O
transcription	O
in	O
Ca3	B-Cell
/	I-Cell
7	I-Cell
cells	I-Cell
transfected	O
with	O
a	O
luciferase	B-Gene_or_gene_product
reporter	I-Gene_or_gene_product
gene	I-Gene_or_gene_product
under	O
the	O
control	O
of	O
glucocorticoid	O
response	O
element	O
(	O
GRE	O
)	O
,	O
but	O
DMI	B-Simple_chemical
alone	O
did	O
not	O
affect	O
GR	B-Gene_or_gene_product
-	O
mediated	O
transcription	O
.	O

However	O
,	O
DMI	B-Simple_chemical
inhibited	O
FA	B-Simple_chemical
-	O
induced	O
,	O
GR	B-Gene_or_gene_product
-	O
mediated	O
transcription	O
when	O
both	O
agents	O
were	O
given	O
together	O
.	O

Pretreatment	O
with	O
DMI	B-Simple_chemical
followed	O
by	O
combination	O
of	O
DMI	B-Simple_chemical
and	O
FA	B-Simple_chemical
decreased	O
GR	B-Gene_or_gene_product
-	O
mediated	O
transcription	O
more	O
than	O
pretreatment	O
with	O
FA	B-Simple_chemical
.	O

The	O
expression	O
of	O
metallothionein	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
Mt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
gene	O
,	O
which	O
is	O
regulated	O
by	O
GR	B-Gene_or_gene_product
,	O
was	O
induced	O
significantly	O
by	O
the	O
combination	O
of	O
DMI	B-Simple_chemical
and	O
FA	B-Simple_chemical
,	O
and	O
enhanced	O
significantly	O
by	O
pretreatment	O
with	O
FA	B-Simple_chemical
but	O
not	O
DMI	B-Simple_chemical
.	O

DMI	B-Simple_chemical
is	O
suggested	O
to	O
inhibit	O
the	O
growth	O
of	O
Ca3	B-Cell
/	I-Cell
7	I-Cell
cells	I-Cell
and	O
to	O
affect	O
GR	B-Gene_or_gene_product
-	O
mediated	O
transcription	O
.	O

MEK5	B-Gene_or_gene_product
/	O
ERK5	B-Gene_or_gene_product
signaling	O
modulates	O
endothelial	B-Cell
cell	I-Cell
migration	O
and	O
focal	B-Cellular_component
contact	I-Cellular_component
turnover	O
.	O

The	O
formation	O
of	O
new	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
from	O
pre	O
-	O
existing	O
ones	O
requires	O
highly	O
coordinated	O
restructuring	O
of	O
endothelial	B-Cell
cells	I-Cell
(	O
EC	B-Cell
)	O
and	O
the	O
surrounding	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
.	O

Directed	O
EC	B-Cell
migration	O
is	O
a	O
central	O
step	O
in	O
this	O
process	O
and	O
depends	O
on	O
cellular	B-Cell
signaling	O
cascades	O
that	O
initiate	O
and	O
control	O
the	O
structural	O
rearrangements	O
.	O

On	O
the	O
basis	O
of	O
earlier	O
findings	O
that	O
ERK5	B-Gene_or_gene_product
deficiency	O
in	O
mouse	B-Organism
EC	B-Cell
results	O
in	O
massive	O
defects	O
in	O
vessel	B-Multi-tissue_structure
architecture	I-Multi-tissue_structure
,	O
we	O
focused	O
on	O
the	O
impact	O
of	O
the	O
MEK5	B-Gene_or_gene_product
/	O
ERK5	B-Gene_or_gene_product
signaling	O
pathway	O
on	O
EC	B-Cell
migration	O
.	O

Using	O
a	O
retroviral	O
gene	O
transfer	O
approach	O
,	O
we	O
found	O
that	O
constitutive	O
activation	O
of	O
MEK5	B-Gene_or_gene_product
/	O
ERK5	B-Gene_or_gene_product
signaling	O
strongly	O
inhibits	O
EC	B-Cell
migration	O
and	O
results	O
in	O
massive	O
morphological	O
changes	O
.	O

The	O
area	O
covered	O
by	O
spread	O
EC	B-Cell
was	O
dramatically	O
enlarged	O
,	O
accompanied	O
by	O
an	O
increase	O
in	O
focal	B-Cellular_component
contacts	I-Cellular_component
and	O
altered	O
organization	O
of	O
actin	B-Gene_or_gene_product
filaments	B-Cellular_component
.	O

Consequently	O
,	O
cells	B-Cell
were	O
more	O
rigid	O
and	O
show	O
reduced	O
motility	O
.	O

This	O
phenotype	O
was	O
most	O
likely	O
based	O
on	O
decreased	O
focal	B-Cellular_component
contact	I-Cellular_component
turnover	O
caused	O
by	O
reduced	O
expression	O
of	O
p130Cas	B-Gene_or_gene_product
,	O
a	O
key	O
player	O
in	O
directed	O
cell	B-Cell
migration	O
.	O

We	O
demonstrate	O
for	O
the	O
first	O
time	O
that	O
ERK5	B-Gene_or_gene_product
signaling	O
not	O
only	O
is	O
involved	O
in	O
EC	B-Cell
survival	O
and	O
stress	O
response	O
but	O
also	O
controls	O
migration	O
and	O
morphology	O
of	O
EC	B-Cell
.	O

EPOX	B-Simple_chemical
inhibits	O
angiogenesis	O
by	O
degradation	O
of	O
Mcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
through	O
ERK	B-Gene_or_gene_product
inactivation	O
.	O

PURPOSE	O
:	O
Antiangiogenic	O
therapy	O
is	O
considered	O
as	O
an	O
effective	O
strategy	O
for	O
controlling	O
the	O
growth	O
and	O
metastasis	O
of	O
tumors	B-Cancer
.	O

Among	O
a	O
myriad	O
of	O
biological	O
activities	O
described	O
for	O
xanthone	B-Simple_chemical
derivatives	I-Simple_chemical
,	O
the	O
anticancer	B-Cancer
activity	O
is	O
quite	O
remarkable	O
,	O
but	O
the	O
molecular	O
mechanism	O
is	O
not	O
clearly	O
resolved	O
.	O

In	O
the	O
present	O
study	O
,	O
we	O
investigated	O
the	O
antiangiogenic	O
mechanism	O
of	O
3	B-Simple_chemical
,	I-Simple_chemical
6	I-Simple_chemical
-	I-Simple_chemical
di	I-Simple_chemical
(	I-Simple_chemical
2	I-Simple_chemical
,	I-Simple_chemical
3	I-Simple_chemical
-	I-Simple_chemical
epoxypropoxy	I-Simple_chemical
)	I-Simple_chemical
xanthone	I-Simple_chemical
(	O
EPOX	B-Simple_chemical
)	O
,	O
a	O
novel	O
Mcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
targeting	O
drug	O
.	O

EXPERIMENTAL	O
DESIGN	O
:	O
To	O
evaluate	O
the	O
antiangiogenic	O
activity	O
of	O
EPOX	B-Simple_chemical
,	O
we	O
did	O
cell	B-Cell
viability	O
,	O
cell	B-Cell
cycle	O
,	O
tube	B-Tissue
formation	O
assay	O
in	O
vitro	O
,	O
and	O
Matrigel	O
plug	O
assay	O
in	O
vivo	O
.	O

To	O
evaluate	O
the	O
effect	O
of	O
EPOX	B-Simple_chemical
on	O
the	O
endothelial	B-Cell
signaling	O
pathway	O
,	O
we	O
did	O
immunoblotting	O
,	O
immunoprecipitation	O
,	O
and	O
immunofluorescence	O
analysis	O
.	O

Intracellular	B-Immaterial_anatomical_entity
glutathione	B-Simple_chemical
levels	O
were	O
determined	O
with	O
the	O
use	O
of	O
monochlorobimane	B-Simple_chemical
,	O
a	O
glutathione	B-Simple_chemical
-	O
specific	O
probe	O
.	O

RESULTS	O
:	O
EPOX	B-Simple_chemical
induced	O
endothelial	B-Cell
cell	I-Cell
apoptosis	O
in	O
association	O
with	O
proteasome	O
-	O
dependent	O
Mcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
degradation	O
.	O

Down	O
-	O
regulation	O
of	O
Mcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
resulted	O
in	O
an	O
increase	O
in	O
Mcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
free	O
Bim	B-Gene_or_gene_product
,	O
activation	O
of	O
Bax	B-Gene_or_gene_product
,	O
and	O
then	O
signaling	O
of	O
mitochondria	B-Cellular_component
-	O
mediated	O
apoptosis	O
.	O

Additionally	O
,	O
glutathione	B-Simple_chemical
depletion	O
and	O
extracellular	B-Gene_or_gene_product
signal	I-Gene_or_gene_product
-	I-Gene_or_gene_product
regulated	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
(	O
ERK	B-Gene_or_gene_product
)	O
inactivation	O
was	O
observed	O
in	O
EPOX	B-Simple_chemical
-	O
treated	O
cells	B-Cell
.	O

Glutathione	B-Simple_chemical
supplementation	O
reversed	O
the	O
inhibitory	O
effects	O
of	O
EPOX	B-Simple_chemical
on	O
ERK	B-Gene_or_gene_product
,	O
which	O
increases	O
the	O
phosphorylation	O
of	O
Mcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
at	O
T	O
(	O
163	O
.	O
)	O
Overexpression	O
of	O
mitogen	B-Gene_or_gene_product
-	I-Gene_or_gene_product
activated	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
/	I-Gene_or_gene_product
ERK	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
(	O
MEK	B-Gene_or_gene_product
)	O
partially	O
reversed	O
the	O
effect	O
of	O
EPOX	B-Simple_chemical
on	O
Mcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
dephosphorylation	O
,	O
ubiquitination	O
,	O
and	O
degradation	O
,	O
further	O
implicating	O
ERK	B-Gene_or_gene_product
in	O
the	O
regulation	O
of	O
Mcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
stability	O
.	O

CONCLUSIONS	O
:	O
This	O
study	O
provides	O
evidence	O
that	O
EPOX	B-Simple_chemical
induces	O
glutathione	B-Simple_chemical
depletion	O
,	O
ERK	B-Gene_or_gene_product
inactivation	O
,	O
and	O
Mcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
degradation	O
on	O
endothelial	B-Cell
cells	I-Cell
,	O
which	O
leads	O
to	O
inhibition	O
of	O
angiogenesis	O
.	O

Our	O
results	O
suggest	O
that	O
EPOX	B-Simple_chemical
is	O
a	O
novel	O
antiangiogenic	O
agent	O
,	O
making	O
it	O
a	O
promising	O
lead	O
compound	O
for	O
further	O
development	O
in	O
the	O
treatment	O
of	O
angiogenesis	O
-	O
related	O
pathologies	O
.	O

Targeting	O
angiogenesis	O
:	O
progress	O
with	O
anti	B-Gene_or_gene_product
-	I-Gene_or_gene_product
VEGF	I-Gene_or_gene_product
treatment	O
with	O
large	O
molecules	O
.	O

Angiogenesis	O
-	O
-	O
one	O
of	O
the	O
hallmarks	O
of	O
cancer	B-Cancer
-	O
-	O
has	O
emerged	O
as	O
a	O
valid	O
therapeutic	O
target	O
in	O
oncology	O
.	O

The	O
VEGF	B-Gene_or_gene_product
system	O
represents	O
a	O
key	O
mediator	O
of	O
tumor	B-Cancer
-	O
initiated	O
angiogenesis	O
and	O
the	O
first	O
target	O
of	O
antiangiogenesis	O
agents	O
introduced	O
in	O
clinical	O
practice	O
.	O

Although	O
anti	B-Gene_or_gene_product
-	I-Gene_or_gene_product
VEGF	I-Gene_or_gene_product
therapies	O
have	O
clearly	O
demonstrated	O
antitumor	B-Cancer
efficacy	O
in	O
various	O
malignancies	B-Cancer
,	O
especially	O
when	O
combined	O
with	O
conventional	O
cytotoxic	O
chemotherapy	O
,	O
their	O
mechanism	O
of	O
action	O
is	O
not	O
fully	O
understood	O
.	O

This	O
Review	O
will	O
discuss	O
the	O
rationale	O
for	O
using	O
antiangiogenic	O
compounds	O
and	O
will	O
focus	O
on	O
large	O
molecules	O
,	O
such	O
as	O
antibodies	O
,	O
that	O
target	O
the	O
VEGF	B-Gene_or_gene_product
system	O
.	O

Clinical	O
data	O
on	O
bevacizumab	B-Simple_chemical
is	O
discussed	O
in	O
detail	O
.	O

Predictive	O
markers	O
for	O
anti	B-Gene_or_gene_product
-	I-Gene_or_gene_product
VEGF	I-Gene_or_gene_product
agents	O
have	O
not	O
yet	O
been	O
identified	O
and	O
questions	O
regarding	O
the	O
usefulness	O
of	O
bevacizumab	B-Simple_chemical
in	O
the	O
adjuvant	O
setting	O
as	O
well	O
as	O
its	O
continued	O
use	O
beyond	O
progression	O
remain	O
unanswered	O
,	O
in	O
spite	O
of	O
negative	O
data	O
on	O
bevacizumab	B-Simple_chemical
in	O
treating	O
patients	B-Organism
with	O
adjuvant	O
colon	B-Cancer
cancer	I-Cancer
.	O

Nonetheless	O
,	O
anti	B-Gene_or_gene_product
-	I-Gene_or_gene_product
VEGF	I-Gene_or_gene_product
therapy	O
has	O
enhanced	O
the	O
arsenal	O
of	O
anticancer	B-Cancer
therapies	O
and	O
has	O
provided	O
new	O
insights	O
into	O
the	O
biology	O
of	O
malignancy	B-Cancer
.	O

[	O
Ambiguity	O
role	O
of	O
neutrophils	B-Cell
in	O
oncogenesis	O
]	O

The	O
review	O
is	O
focused	O
on	O
the	O
participation	O
of	O
polymorphonuclear	B-Cell
granulocytes	I-Cell
(	O
neutrophils	B-Cell
)	O
in	O
development	O
and	O
spreading	O
of	O
a	O
tumor	B-Cancer
.	O

We	O
consider	O
both	O
the	O
well	O
known	O
functions	O
of	O
neutrophils	B-Cell
(	O
degranulation	O
,	O
production	O
of	O
reactive	B-Simple_chemical
oxygen	I-Simple_chemical
species	I-Simple_chemical
(	O
ROS	B-Simple_chemical
)	O
)	O
and	O
the	O
recently	O
shown	O
one	O
(	O
presentation	O
of	O
an	O
antigene	O
)	O
.	O

The	O
special	O
attention	O
is	O
focused	O
on	O
the	O
ambiguity	O
of	O
the	O
neutrophil	B-Cell
role	O
in	O
oncogenesis	O
.	O

The	O
dominant	O
view	O
is	O
that	O
neutrophils	B-Cell
display	O
exclusively	O
antitumor	B-Cancer
properties	O
.	O

The	O
update	O
information	O
testifies	O
about	O
protumoral	B-Cancer
activity	O
of	O
neutrophils	B-Cell
:	O
they	O
migrate	O
to	O
a	O
tumor	B-Cancer
and	O
promote	O
angiogenesis	O
and	O
metastasis	O
at	O
late	O
stages	O
of	O
the	O
tumor	B-Cancer
.	O

It	O
is	O
interesting	O
that	O
certain	O
components	O
of	O
neutrophil	B-Cell
cytotoxic	O
arsenal	O
(	O
ROS	B-Simple_chemical
,	O
cytokines	O
,	O
specific	O
enzymes	O
)	O
participate	O
both	O
in	O
antitumoral	B-Cancer
defenses	O
of	O
an	O
organism	O
and	O
protumoral	B-Cancer
activity	O
.	O

Endothelial	B-Cell
cell	I-Cell
migration	O
and	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
expression	O
are	O
the	O
result	O
of	O
loss	O
of	O
breast	B-Tissue
tissue	I-Tissue
polarity	O
.	O

Recruiting	O
a	O
new	O
blood	B-Organism_substance
supply	O
is	O
a	O
rate	O
-	O
limiting	O
step	O
in	O
tumor	B-Cancer
progression	O
.	O

In	O
a	O
three	O
-	O
dimensional	O
model	O
of	O
breast	B-Organism_subdivision
carcinogenesis	O
,	O
disorganized	O
,	O
proliferative	O
transformed	O
breast	B-Cell
epithelial	I-Cell
cells	I-Cell
express	O
significantly	O
higher	O
expression	O
of	O
angiogenic	O
genes	O
compared	O
with	O
their	O
polarized	O
,	O
growth	O
-	O
arrested	O
nonmalignant	B-Cell
counterparts	I-Cell
.	O

Elevated	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
secretion	O
by	O
malignant	B-Cell
cells	I-Cell
enhanced	O
recruitment	O
of	O
endothelial	B-Cell
cells	I-Cell
(	O
EC	B-Cell
)	O
in	O
heterotypic	O
cocultures	O
.	O

Significantly	O
,	O
phenotypic	O
reversion	O
of	O
malignant	B-Cell
cells	I-Cell
via	O
reexpression	O
of	O
HoxD10	B-Gene_or_gene_product
,	O
which	O
is	O
lost	O
in	O
malignant	O
progression	O
,	O
significantly	O
attenuated	O
VEGF	B-Gene_or_gene_product
expression	O
in	O
a	O
hypoxia	B-Gene_or_gene_product
-	I-Gene_or_gene_product
inducible	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
-	O
independent	O
fashion	O
and	O
reduced	O
EC	B-Cell
migration	O
.	O

This	O
was	O
due	O
primarily	O
to	O
restoring	O
polarity	O
:	O
forced	O
proliferation	O
of	O
polarized	O
,	O
nonmalignant	B-Cell
cells	I-Cell
did	O
not	O
induce	O
VEGF	B-Gene_or_gene_product
expression	O
and	O
EC	B-Cell
recruitment	O
,	O
whereas	O
disrupting	O
the	O
architecture	O
of	O
growth	O
-	O
arrested	O
,	O
reverted	O
cells	B-Cell
did	O
.	O

These	O
data	O
show	O
that	O
disrupting	O
cytostructure	B-Cellular_component
activates	O
the	O
angiogenic	O
switch	O
even	O
in	O
the	O
absence	O
of	O
proliferation	O
and	O
/	O
or	O
hypoxia	O
and	O
restoring	O
organization	O
of	O
malignant	B-Cancer
clusters	I-Cancer
reduces	O
VEGF	B-Gene_or_gene_product
expression	O
and	O
EC	B-Cell
activation	O
to	O
levels	O
found	O
in	O
quiescent	O
nonmalignant	B-Tissue
epithelium	I-Tissue
.	O

These	O
data	O
confirm	O
the	O
importance	O
of	O
tissue	B-Tissue
architecture	O
and	O
polarity	O
in	O
malignant	O
progression	O
.	O

Metronomic	O
5	B-Simple_chemical
-	I-Simple_chemical
fluorouracil	I-Simple_chemical
,	O
oxaliplatin	B-Simple_chemical
and	O
irinotecan	B-Simple_chemical
in	O
colorectal	B-Cancer
cancer	I-Cancer
.	O

Metronomic	O
chemotherapy	O
(	O
the	O
frequent	O
,	O
long	O
term	O
,	O
low	O
dose	O
administration	O
of	O
chemotherapeutic	O
drugs	O
)	O
is	O
a	O
promising	O
therapy	O
because	O
it	O
enhances	O
the	O
anti	O
-	O
endothelial	B-Cell
activity	O
of	O
conventional	O
chemotherapeutics	O
,	O
but	O
with	O
lower	O
or	O
no	O
toxic	O
effects	O
compared	O
to	O
maximum	O
tolerated	O
dose	O
administration	O
.	O

The	O
aims	O
of	O
the	O
present	O
study	O
were	O
to	O
compare	O
,	O
in	O
vitro	O
and	O
in	O
vivo	O
,	O
the	O
antiangiogenic	O
and	O
antitumor	B-Cancer
activities	O
of	O
metronomic	O
irinotecan	B-Simple_chemical
(	O
CPT	B-Simple_chemical
-	I-Simple_chemical
11	I-Simple_chemical
)	O
,	O
oxaliplatin	B-Simple_chemical
(	O
L	B-Simple_chemical
-	I-Simple_chemical
OHP	I-Simple_chemical
)	O
and	O
5	B-Simple_chemical
-	I-Simple_chemical
fluorouracil	I-Simple_chemical
(	O
5	B-Simple_chemical
-	I-Simple_chemical
FU	I-Simple_chemical
)	O
in	O
colorectal	B-Cancer
cancer	I-Cancer
and	O
to	O
investigate	O
the	O
metronomic	O
combination	O
of	O
these	O
drugs	O
.	O

In	O
vitro	O
cell	B-Cell
proliferation	O
,	O
combination	O
studies	O
and	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
secretion	O
analyses	O
were	O
performed	O
on	O
endothelial	B-Cell
(	I-Cell
HMVEC	I-Cell
-	I-Cell
d	I-Cell
)	I-Cell
and	O
colorectal	B-Cell
cancer	I-Cell
(	I-Cell
HT	I-Cell
-	I-Cell
29	I-Cell
)	I-Cell
cells	I-Cell
exposed	O
for	O
144	O
h	O
to	O
metronomic	O
concentrations	O
of	O
SN	B-Simple_chemical
-	I-Simple_chemical
38	I-Simple_chemical
,	O
the	O
active	O
metabolite	O
of	O
CPT	B-Simple_chemical
-	I-Simple_chemical
11	I-Simple_chemical
,	O
L	B-Simple_chemical
-	I-Simple_chemical
OHP	I-Simple_chemical
and	O
5	B-Simple_chemical
-	I-Simple_chemical
FU	I-Simple_chemical
.	O

HT	B-Cell
-	I-Cell
29	I-Cell
human	B-Organism
colorectal	B-Cancer
cancer	I-Cancer
xenograft	I-Cancer
model	O
was	O
used	O
and	O
tumour	B-Cancer
growth	O
,	O
microvessel	B-Tissue
density	O
and	O
VEGF	B-Gene_or_gene_product
quantification	O
were	O
performed	O
in	O
tumours	B-Cancer
after	O
the	O
administration	O
of	O
metronomic	O
CPT	B-Simple_chemical
-	I-Simple_chemical
11	I-Simple_chemical
,	O
L	B-Simple_chemical
-	I-Simple_chemical
OHP	I-Simple_chemical
,	O
5	B-Simple_chemical
-	I-Simple_chemical
FU	I-Simple_chemical
and	O
their	O
simultaneous	O
combination	O
.	O

Low	O
concentrations	O
of	O
SN	B-Simple_chemical
-	I-Simple_chemical
38	I-Simple_chemical
,	O
but	O
not	O
5	B-Simple_chemical
-	I-Simple_chemical
FU	I-Simple_chemical
and	O
L	B-Simple_chemical
-	I-Simple_chemical
OHP	I-Simple_chemical
,	O
preferentially	O
inhibited	O
endothelial	B-Cell
cell	I-Cell
proliferation	O
.	O

Simultaneous	O
and	O
continuous	O
exposure	O
of	O
HT	B-Cell
-	I-Cell
29	I-Cell
and	O
HMVEC	B-Cell
-	I-Cell
d	I-Cell
cells	I-Cell
to	O
low	O
concentrations	O
SN	B-Simple_chemical
-	I-Simple_chemical
38	I-Simple_chemical
+	O
L	B-Simple_chemical
-	I-Simple_chemical
OHP	I-Simple_chemical
+	O
5	B-Simple_chemical
-	I-Simple_chemical
FU	I-Simple_chemical
for	O
144	O
h	O
showed	O
a	O
strong	O
antagonism	O
and	O
an	O
unfavorable	O
dose	O
-	O
reduction	O
index	O
.	O

Moreover	O
,	O
the	O
ternary	O
combination	O
resulted	O
in	O
a	O
significant	O
increase	O
of	O
VEGF	B-Gene_or_gene_product
secretion	O
in	O
HT	B-Cell
-	I-Cell
29	I-Cell
cancer	I-Cell
cells	I-Cell
.	O

In	O
a	O
xenograft	B-Cancer
model	O
metronomic	O
CPT	B-Simple_chemical
-	I-Simple_chemical
11	I-Simple_chemical
,	O
but	O
not	O
5	B-Simple_chemical
-	I-Simple_chemical
FU	I-Simple_chemical
and	O
L	B-Simple_chemical
-	I-Simple_chemical
OHP	I-Simple_chemical
,	O
significantly	O
inhibits	O
HT	B-Cancer
-	I-Cancer
29	I-Cancer
tumor	I-Cancer
growth	O
and	O
microvessel	B-Tissue
density	O
in	O
the	O
absence	O
of	O
toxicity	O
.	O

On	O
the	O
contrary	O
,	O
metronomic	O
5	B-Simple_chemical
-	I-Simple_chemical
FU	I-Simple_chemical
+	O
L	B-Simple_chemical
-	I-Simple_chemical
OHP	I-Simple_chemical
+	O
CPT	B-Simple_chemical
-	I-Simple_chemical
11	I-Simple_chemical
therapy	O
did	O
not	O
affect	O
the	O
microvascular	B-Tissue
count	O
.	O

The	O
metronomic	O
concept	O
might	O
not	O
universally	O
apply	O
to	O
every	O
cytotoxic	O
drug	O
in	O
colorectal	B-Cancer
cancer	I-Cancer
and	O
metronomic	O
combination	O
regimens	O
should	O
be	O
used	O
with	O
caution	O
.	O

A	O
key	O
role	O
for	O
the	O
integrin	B-Gene_or_gene_product
alpha2beta1	I-Gene_or_gene_product
in	O
experimental	O
and	O
developmental	O
angiogenesis	O
.	O

The	O
alpha2beta1	B-Gene_or_gene_product
integrin	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
plays	O
a	O
key	O
role	O
in	O
angiogenesis	O
.	O

Here	O
we	O
investigated	O
the	O
effects	O
of	O
small	O
molecule	O
inhibitors	O
(	O
SMIs	O
)	O
designed	O
to	O
disrupt	O
integrin	B-Gene_or_gene_product
alpha2	I-Gene_or_gene_product
I	O
or	O
beta1	B-Gene_or_gene_product
I	O
-	O
like	O
domain	O
function	O
on	O
angiogenesis	O
.	O

In	O
unchallenged	O
endothelial	B-Cell
cells	I-Cell
,	O
fibrillar	B-Gene_or_gene_product
collagen	I-Gene_or_gene_product
induced	O
robust	O
capillary	B-Tissue
morphogenesis	O
.	O

In	O
contrast	O
,	O
tube	B-Tissue
formation	O
was	O
significantly	O
reduced	O
by	O
SMI496	B-Simple_chemical
,	O
a	O
beta1	B-Gene_or_gene_product
I	O
-	O
like	O
domain	O
inhibitor	O
and	O
by	O
function	O
-	O
blocking	O
anti	B-Simple_chemical
-	I-Simple_chemical
alpha2beta1	I-Simple_chemical
but	O
not	O
-	B-Simple_chemical
alpha1beta1	I-Simple_chemical
antibodies	I-Simple_chemical
.	O

Endothelial	B-Cell
cells	I-Cell
bound	O
fluorescein	B-Simple_chemical
-	O
labeled	O
collagen	B-Gene_or_gene_product
I	I-Gene_or_gene_product
fibrils	I-Gene_or_gene_product
,	O
an	O
interaction	O
specifically	O
inhibited	O
by	O
SMI496	B-Simple_chemical
.	O

Moreover	O
,	O
SMI496	B-Simple_chemical
caused	O
cell	B-Cell
retraction	O
and	O
cytoskeletal	O
collapse	O
of	O
endothelial	B-Cell
cells	I-Cell
as	O
well	O
as	O
delayed	O
endothelial	B-Cell
cell	I-Cell
wound	B-Pathological_formation
healing	O
.	O

SMI	O
activities	O
were	O
examined	O
in	O
vivo	O
by	O
supplementing	O
the	O
growth	O
medium	O
of	O
zebrafish	B-Organism
embryos	B-Developing_anatomical_structure
expressing	O
green	B-Gene_or_gene_product
fluorescent	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
under	O
the	O
control	O
of	O
the	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
promoter	O
.	O

SMI496	B-Simple_chemical
,	O
but	O
not	O
a	O
control	O
compound	O
,	O
interfered	O
with	O
angiogenesis	O
in	O
vivo	O
by	O
reversibly	O
inhibiting	O
sprouting	O
from	O
the	O
axial	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
.	O

We	O
further	O
characterized	O
zebrafish	B-Organism
alpha2	B-Gene_or_gene_product
integrin	I-Gene_or_gene_product
and	O
discovered	O
that	O
this	O
integrin	B-Gene_or_gene_product
is	O
highly	O
conserved	O
,	O
especially	O
the	O
I	O
domain	O
.	O

Notably	O
,	O
a	O
similar	O
vascular	B-Multi-tissue_structure
phenotype	O
was	O
induced	O
by	O
morpholino	O
-	O
mediated	O
knockdown	O
of	O
the	O
integrin	B-Gene_or_gene_product
alpha2	I-Gene_or_gene_product
subunit	O
.	O

By	O
live	O
videomicroscopy	O
,	O
we	O
confirmed	O
that	O
the	O
vessels	B-Multi-tissue_structure
were	O
largely	O
nonfunctional	O
in	O
the	O
absence	O
of	O
alpha2beta1	B-Gene_or_gene_product
integrin	I-Gene_or_gene_product
.	O

Collectively	O
,	O
our	O
results	O
provide	O
strong	O
biochemical	O
and	O
genetic	O
evidence	O
of	O
a	O
central	O
role	O
for	O
alpha2beta1	B-Gene_or_gene_product
integrin	I-Gene_or_gene_product
in	O
experimental	O
and	O
developmental	O
angiogenesis	O
.	O

Islet	B-Cell
endothelial	I-Cell
activation	O
and	O
oxidative	O
stress	O
gene	O
expression	O
is	O
reduced	O
by	O
IL	B-Simple_chemical
-	I-Simple_chemical
1Ra	I-Simple_chemical
treatment	O
in	O
the	O
type	O
2	O
diabetic	O
GK	B-Organism
rat	I-Organism
.	O

BACKGROUND	O
:	O
Inflammation	O
followed	O
by	O
fibrosis	O
is	O
a	O
component	O
of	O
islet	B-Multi-tissue_structure
dysfunction	O
in	O
both	O
rodent	O
and	O
human	B-Organism
type	O
2	O
diabetes	O
.	O

Because	O
islet	B-Multi-tissue_structure
inflammation	O
may	O
originate	O
from	O
endothelial	B-Cell
cells	I-Cell
,	O
we	O
assessed	O
the	O
expression	O
of	O
selected	O
genes	O
involved	O
in	O
endothelial	B-Cell
cell	I-Cell
activation	O
in	O
islets	B-Multi-tissue_structure
from	O
a	O
spontaneous	O
model	O
of	O
type	O
2	O
diabetes	O
,	O
the	O
Goto	B-Organism
-	I-Organism
Kakizaki	I-Organism
(	I-Organism
GK	I-Organism
)	I-Organism
rat	I-Organism
.	O

We	O
also	O
examined	O
islet	B-Tissue
endotheliuml	I-Tissue
/	O
oxidative	O
stress	O
(	O
OS	O
)	O
/	O
inflammation	O
-	O
related	O
gene	O
expression	O
,	O
islet	B-Multi-tissue_structure
vascularization	O
and	O
fibrosis	O
after	O
treatment	O
with	O
the	O
interleukin	B-Simple_chemical
-	I-Simple_chemical
1	I-Simple_chemical
(	I-Simple_chemical
IL	I-Simple_chemical
-	I-Simple_chemical
1	I-Simple_chemical
)	I-Simple_chemical
receptor	I-Simple_chemical
antagonist	I-Simple_chemical
(	O
IL	B-Simple_chemical
-	I-Simple_chemical
1Ra	I-Simple_chemical
)	O
.	O

METHODOLOGY	O
/	O
PRINCIPAL	O
FINDINGS	O
:	O
Gene	O
expression	O
was	O
analyzed	O
by	O
quantitative	O
RT	O
-	O
PCR	O
on	O
islets	B-Multi-tissue_structure
isolated	O
from	O
10	O
-	O
week	O
-	O
old	O
diabetic	O
GK	B-Organism
and	O
control	O
Wistar	B-Organism
rats	I-Organism
.	O

Furthermore	O
,	O
GK	B-Organism
rats	I-Organism
were	O
treated	O
s	O
.	O
c	O
twice	O
daily	O
with	O
IL	B-Simple_chemical
-	I-Simple_chemical
1Ra	I-Simple_chemical
(	O
Kineret	B-Simple_chemical
,	O
Amgen	O
,	O
100	O
mg	O
/	O
kg	O
/	O
day	O
)	O
or	O
saline	B-Simple_chemical
,	O
from	O
4	O
weeks	O
of	O
age	O
onwards	O
(	O
onset	O
of	O
diabetes	O
)	O
.	O

Four	O
weeks	O
later	O
,	O
islet	B-Multi-tissue_structure
gene	O
analysis	O
and	O
pancreas	B-Organ
immunochemistry	O
were	O
performed	O
.	O

Thirty	O
-	O
two	O
genes	O
were	O
selected	O
encoding	O
molecules	O
involved	O
in	O
endothelial	B-Cell
cell	I-Cell
activation	O
,	O
particularly	O
fibrinolysis	O
,	O
vascular	B-Multi-tissue_structure
tone	O
,	O
OS	O
,	O
angiogenesis	O
and	O
also	O
inflammation	O
.	O

All	O
genes	O
except	O
those	O
encoding	O
angiotensinogen	B-Gene_or_gene_product
and	O
epoxide	B-Gene_or_gene_product
hydrolase	I-Gene_or_gene_product
(	O
that	O
were	O
decreased	O
)	O
,	O
and	O
12	B-Gene_or_gene_product
-	I-Gene_or_gene_product
lipoxygenase	I-Gene_or_gene_product
and	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
that	O
showed	O
no	O
change	O
)	O
,	O
were	O
significantly	O
up	O
-	O
regulated	O
in	O
GK	B-Organism
islets	B-Multi-tissue_structure
.	O

After	O
IL	B-Simple_chemical
-	I-Simple_chemical
1Ra	I-Simple_chemical
treatment	O
of	O
GK	B-Organism
rats	I-Organism
in	O
vivo	O
,	O
most	O
selected	O
genes	O
implied	O
in	O
endothelium	B-Tissue
/	O
OS	O
/	O
immune	B-Cell
cells	I-Cell
/	O
fibrosis	O
were	O
significantly	O
down	O
-	O
regulated	O
.	O

IL	B-Simple_chemical
-	I-Simple_chemical
1Ra	I-Simple_chemical
also	O
improved	O
islet	B-Multi-tissue_structure
vascularization	O
,	O
reduced	O
fibrosis	O
and	O
ameliorated	O
glycemia	O
.	O

CONCLUSIONS	O
/	O
SIGNIFICANCE	O
:	O
GK	B-Organism
rat	I-Organism
islets	B-Multi-tissue_structure
have	O
increased	O
mRNA	O
expression	O
of	O
markers	O
of	O
early	O
islet	B-Cell
endothelial	I-Cell
cell	I-Cell
activation	O
,	O
possibly	O
triggered	O
by	O
several	O
metabolic	O
factors	O
,	O
and	O
also	O
some	O
defense	O
mechanisms	O
.	O

The	O
beneficial	O
effect	O
of	O
IL	B-Simple_chemical
-	I-Simple_chemical
1Ra	I-Simple_chemical
on	O
most	O
islet	B-Cell
endothelial	I-Cell
/	O
OS	O
/	O
immune	B-Cell
cells	I-Cell
/	O
fibrosis	O
parameters	O
analyzed	O
highlights	O
a	O
major	O
endothelial	B-Cell
-	O
related	O
role	O
for	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
in	O
GK	B-Organism
islet	B-Multi-tissue_structure
alterations	O
.	O

Thus	O
,	O
metabolically	O
-	O
altered	O
islet	B-Tissue
endothelium	I-Tissue
might	O
affect	O
the	O
beta	B-Cell
-	I-Cell
cell	I-Cell
microenvironment	O
and	O
contribute	O
to	O
progressive	O
type	O
2	O
diabetic	O
beta	B-Cell
-	I-Cell
cell	I-Cell
dysfunction	O
in	O
GK	B-Organism
rats	I-Organism
.	O

Counteracting	O
islet	B-Cell
endothelial	I-Cell
cell	I-Cell
inflammation	O
might	O
be	O
one	O
way	O
to	O
ameliorate	O
/	O
prevent	O
beta	B-Cell
-	I-Cell
cell	I-Cell
dysfunction	O
in	O
type	O
2	O
diabetes	O
.	O

AVE8062	B-Simple_chemical
:	O
a	O
new	O
combretastatin	B-Simple_chemical
derivative	O
vascular	B-Multi-tissue_structure
disrupting	O
agent	O
.	O

Angiogenesis	O
has	O
an	O
essential	O
role	O
in	O
promoting	O
and	O
supporting	O
tumor	B-Cancer
growth	O
and	O
it	O
is	O
an	O
important	O
therapeutic	O
target	O
.	O

The	O
tumor	B-Multi-tissue_structure
vascular	I-Multi-tissue_structure
network	I-Multi-tissue_structure
is	O
the	O
result	O
of	O
pro	O
-	O
angiogenic	O
and	O
inhibitory	O
factors	O
as	O
well	O
as	O
of	O
the	O
interaction	O
between	O
endothelial	B-Cell
cells	I-Cell
and	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
.	O

Different	O
antiangiogenic	O
therapeutics	O
have	O
been	O
developed	O
to	O
improve	O
tumor	B-Cancer
control	O
through	O
vascular	B-Simple_chemical
-	I-Simple_chemical
targeting	I-Simple_chemical
agents	I-Simple_chemical
(	O
VTA	B-Simple_chemical
)	O
.	O

VTAs	B-Simple_chemical
can	O
be	O
divided	O
into	O
two	O
groups	O
:	O
antiangiogenic	B-Simple_chemical
agents	I-Simple_chemical
and	O
vascular	B-Simple_chemical
-	I-Simple_chemical
disrupting	I-Simple_chemical
agents	I-Simple_chemical
(	O
VDAs	B-Simple_chemical
)	O
.	O

VTAs	B-Simple_chemical
inhibit	O
specific	O
factors	O
required	O
to	O
induce	O
and	O
direct	O
the	O
angiogenic	O
process	O
,	O
with	O
major	O
activity	O
against	O
small	O
tumor	B-Cancer
masses	I-Cancer
and	O
at	O
the	O
tumor	B-Tissue
periphery	I-Tissue
,	O
encompassing	O
monoclonal	O
antibodies	O
and	O
small	O
molecules	O
inhibitors	O
of	O
the	O
tyrosine	O
kinase	O
domain	O
of	O
the	O
VEGF	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
.	O

VDAs	B-Simple_chemical
specifically	O
target	O
and	O
destroy	O
well	O
-	O
established	O
tumor	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
with	O
ischemia	O
and	O
destruction	O
of	O
large	O
masses	B-Cancer
with	O
central	O
hemorrhagic	O
necrosis	O
and	O
survival	O
of	O
a	O
thin	O
peripheral	B-Cancer
tumor	I-Cancer
layer	I-Cancer
.	O

VDAs	B-Simple_chemical
can	O
be	O
divided	O
into	O
biologics	O
,	O
such	O
as	O
ligand	O
-	O
based	O
,	O
and	O
small	O
-	O
molecule	O
agents	O
;	O
this	O
second	O
group	O
includes	O
small	B-Simple_chemical
-	I-Simple_chemical
molecule	I-Simple_chemical
VDAs	I-Simple_chemical
like	O
flavonoids	B-Simple_chemical
,	O
such	O
as	O
5	B-Simple_chemical
,	I-Simple_chemical
6	I-Simple_chemical
-	I-Simple_chemical
dimethylxanthenone	I-Simple_chemical
-	I-Simple_chemical
4	I-Simple_chemical
-	I-Simple_chemical
acetic	I-Simple_chemical
acid	I-Simple_chemical
(	O
DMXAA	B-Simple_chemical
)	O
,	O
and	O
microtubule	B-Simple_chemical
-	I-Simple_chemical
destabilizing	I-Simple_chemical
agents	I-Simple_chemical
.	O

In	O
this	O
review	O
we	O
will	O
discuss	O
the	O
mechanism	O
of	O
action	O
,	O
as	O
well	O
as	O
the	O
preclinical	O
and	O
clinical	O
results	O
,	O
of	O
one	O
of	O
the	O
most	O
promising	O
antitubulin	B-Simple_chemical
agents	I-Simple_chemical
:	O
the	O
combretastatin	B-Simple_chemical
A4	I-Simple_chemical
-	I-Simple_chemical
phosphate	I-Simple_chemical
derivative	I-Simple_chemical
,	O
AVE8062A	B-Simple_chemical
.	O

Targeting	O
TNF	B-Gene_or_gene_product
for	O
Treatment	O
of	O
Cancer	B-Cancer
and	O
Autoimmunity	O
.	O

Tumor	B-Gene_or_gene_product
necrosis	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
(	O
TNF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
)	O
was	O
first	O
isolated	O
two	O
decades	O
ago	O
as	O
a	O
macrophageproduced	O
protein	O
that	O
can	O
effectively	O
kill	O
tumor	B-Cell
cells	I-Cell
.	O

TNF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
is	O
also	O
an	O
essential	O
component	O
of	O
the	O
immune	O
system	O
and	O
is	O
required	O
for	O
hematopoiesis	O
,	O
for	O
protection	O
from	O
bacterial	O
infection	O
and	O
for	O
immune	B-Cell
cell	I-Cell
-	O
mediated	O
cytotoxicity	O
.	O

Extensive	O
research	O
,	O
however	O
,	O
has	O
revealed	O
that	O
TNF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
is	O
one	O
of	O
the	O
major	O
players	O
in	O
tumor	B-Cancer
initiation	O
,	O
proliferation	O
,	O
invasion	O
,	O
angiogenesis	O
and	O
metastasis	O
.	O

The	O
proinflammatory	O
activities	O
link	O
TNF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
with	O
a	O
wide	O
variety	O
of	O
autoimmune	O
diseases	O
,	O
including	O
psoriasis	O
,	O
inflammatory	O
bowel	B-Organ
disease	O
,	O
rheumatoid	O
arthritis	O
,	O
systemic	O
sclerosis	O
,	O
systemic	O
lupus	O
erythematosus	O
,	O
multiple	O
sclerosis	O
,	O
diabetes	O
and	O
ankylosing	O
spondylitis	O
.	O

Systemic	O
inhibitors	O
of	O
TNF	B-Gene_or_gene_product
such	O
as	O
etanercept	B-Simple_chemical
(	O
Enbrel	B-Simple_chemical
)	O
(	O
a	O
soluble	O
TNF	B-Gene_or_gene_product
receptor	O
)	O
and	O
infliximab	B-Simple_chemical
(	O
Remicade	B-Simple_chemical
)	O
and	O
adalimumab	B-Simple_chemical
(	O
Humira	B-Simple_chemical
)	O
(	O
anti	B-Gene_or_gene_product
-	I-Gene_or_gene_product
TNF	I-Gene_or_gene_product
antibodies	I-Gene_or_gene_product
)	O
have	O
been	O
approved	O
for	O
the	O
treatment	O
inflammatory	O
bowel	B-Organ
disease	O
,	O
psoriasis	O
and	O
rheumatoid	O
arthritis	O
.	O

These	O
drugs	O
,	O
however	O
,	O
exhibit	O
severe	O
side	O
effects	O
and	O
are	O
expensive	O
.	O

Hence	O
orally	O
active	O
blockers	O
of	O
TNF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
that	O
are	O
safe	O
,	O
efficacious	O
and	O
inexpensive	O
are	O
urgently	O
needed	O
.	O

Numerous	O
products	O
from	O
fruits	B-Organism_subdivision
,	O
vegetable	B-Organism_subdivision
and	O
traditional	O
medicinal	O
plants	O
have	O
been	O
described	O
which	O
can	O
suppress	O
TNF	B-Gene_or_gene_product
expression	O
and	O
TNF	B-Gene_or_gene_product
signaling	O
but	O
their	O
clinical	O
potential	O
is	O
yet	O
uncertain	O
.	O

[	O
Effect	O
of	O
osteopontin	B-Gene_or_gene_product
silencing	O
by	O
lentivirus	B-Organism
-	O
mediated	O
delivery	O
of	O
siRNA	O
on	O
glioma	B-Cell
cell	I-Cell
invasion	O
and	O
apoptosis	O
]	O
.	O

OBJECTIVE	O
:	O
To	O
investigate	O
the	O
effect	O
of	O
osteopontin	B-Gene_or_gene_product
silencing	O
on	O
the	O
invasion	O
and	O
apoptosis	O
of	O
U251	B-Cell
cells	I-Cell
.	O

METHODS	O
:	O
The	O
invasion	O
,	O
apoptosis	O
and	O
levels	O
of	O
uPA	B-Gene_or_gene_product
,	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
were	O
determined	O
by	O
invasion	O
assay	O
,	O
flow	O
cytometry	O
,	O
Western	O
blot	O
and	O
real	O
-	O
time	O
fluorescence	O
quantitative	O
PCR	O
respectively	O
.	O

RESULTS	O
:	O
Osteopontin	B-Gene_or_gene_product
small	O
interference	O
RNA	O
(	O
siRNA	O
)	O
inhibited	O
osteopontin	B-Gene_or_gene_product
expression	O
and	O
cell	B-Cell
invasion	O
,	O
promoted	O
apoptosis	O
in	O
U251	B-Cell
cells	I-Cell
.	O

In	O
addition	O
,	O
the	O
expression	O
of	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
uPA	B-Gene_or_gene_product
,	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
was	O
decreased	O
,	O
while	O
Bax	B-Gene_or_gene_product
level	O
was	O
elevated	O
.	O

CONCLUSION	O
:	O
Osteopontin	B-Gene_or_gene_product
siRNA	O
can	O
inhibit	O
U251	B-Cell
cells	I-Cell
invasion	O
via	O
the	O
down	O
-	O
regulation	O
of	O
uPA	B-Gene_or_gene_product
,	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
levels	O
,	O
and	O
promote	O
apoptosis	O
through	O
induction	O
of	O
Bax	B-Gene_or_gene_product
expression	O
and	O
inhibition	O
of	O
Bcl	B-Gene_or_gene_product
2	I-Gene_or_gene_product
level	O
.	O

It	O
suggests	O
that	O
osteopontin	B-Gene_or_gene_product
plays	O
an	O
important	O
role	O
in	O
human	B-Organism
glioma	B-Cancer
progression	O
.	O

Magnitude	O
of	O
malate	B-Simple_chemical
-	I-Simple_chemical
aspartate	I-Simple_chemical
reduced	I-Simple_chemical
nicotinamide	I-Simple_chemical
adenine	I-Simple_chemical
dinucleotide	I-Simple_chemical
shuttle	O
activity	O
in	O
intact	O
respiring	O
tumor	B-Cell
cells	I-Cell
.	O

Measurements	O
of	O
respiration	O
,	O
CO2	B-Simple_chemical
and	O
lactate	B-Simple_chemical
production	O
,	O
and	O
changes	O
in	O
the	O
levels	O
of	O
various	O
key	O
metabolites	O
of	O
the	O
glycolytic	O
sequence	O
and	O
tricarboxylic	B-Simple_chemical
acid	I-Simple_chemical
cycle	O
were	O
made	O
on	O
five	O
lines	O
of	O
rodent	B-Organism
ascites	B-Cell
tumor	I-Cell
cells	I-Cell
(	O
two	O
strains	O
of	O
Ehrlich	B-Cell
ascites	I-Cell
tumor	I-Cell
cells	I-Cell
,	O
Krebs	B-Cell
II	I-Cell
carcinoma	I-Cell
,	O
AS	B-Cell
-	I-Cell
30D	I-Cell
carcinoma	I-Cell
,	O
and	O
L1210	B-Cell
cells	I-Cell
)	O
incubated	O
aerobically	O
in	O
the	O
presence	O
of	O
uniformly	O
labeled	O
D	B-Simple_chemical
-	I-Simple_chemical
[	I-Simple_chemical
14C	I-Simple_chemical
]	I-Simple_chemical
glucose	I-Simple_chemical
.	O

From	O
these	O
data	O
,	O
as	O
well	O
as	O
earlier	O
evidence	O
demonstrating	O
that	O
the	O
reduced	O
nicotinamide	B-Simple_chemical
adenine	I-Simple_chemical
dinucleotide	I-Simple_chemical
(	O
NADH	B-Simple_chemical
)	O
shuttle	O
in	O
these	O
cells	B-Cell
requires	O
a	O
transaminase	O
step	O
and	O
is	O
thus	O
identified	O
as	O
the	O
malate	B-Simple_chemical
-	O
aspartate	B-Simple_chemical
shuttle	O
(	O
W	O
.	O
V	O
.	O
V	O
.	O
Greenhouse	O
and	O
A	O
.	O
L	O
.	O
Lehninger	O
,	O
Cancer	O
Res	O
.	O
,	O
36	O
:	O
1392	O
-	O
1396	O
,	O
1976	O
)	O
,	O
metabolic	O
flux	O
diagrams	O
were	O
constructed	O
for	O
the	O
five	O
cell	B-Cell
lines	I-Cell
.	O

These	O
diagrams	O
show	O
the	O
relative	O
rates	O
of	O
glycolysis	O
,	O
the	O
tricarboxylic	B-Simple_chemical
acid	I-Simple_chemical
cycle	O
,	O
electron	B-Simple_chemical
transport	O
,	O
and	O
the	O
malate	B-Simple_chemical
-	O
aspartate	B-Simple_chemical
shuttle	O
in	O
these	O
tumors	B-Cancer
.	O

Large	O
amounts	O
of	O
cytosolic	O
NADH	B-Simple_chemical
were	O
oxidized	O
by	O
the	O
mitochondrial	B-Cellular_component
respiratory	O
chain	O
via	O
the	O
NADH	B-Simple_chemical
shuttle	O
,	O
comprising	O
anywhere	O
from	O
about	O
20	O
to	O
80	O
%	O
of	O
the	O
total	O
flow	O
of	O
reducing	O
equivalents	O
to	O
oxygen	B-Simple_chemical
in	O
these	O
tumors	B-Cancer
.	O

Calculations	O
of	O
the	O
sources	O
of	O
energy	O
for	O
adenosine	B-Simple_chemical
triphosphate	I-Simple_chemical
synthesis	O
indicated	O
that	O
on	O
the	O
average	O
about	O
one	O
-	O
third	O
of	O
the	O
respiratory	O
adenosine	B-Simple_chemical
triphosphate	I-Simple_chemical
is	O
generated	O
by	O
electron	B-Simple_chemical
flow	O
originating	O
from	O
cytosolic	B-Cellular_component
NADH	B-Simple_chemical
via	O
the	O
malate	B-Simple_chemical
-	O
aspartate	B-Simple_chemical
shuttle	O
.	O

AKT2	B-Gene_or_gene_product
is	O
a	O
downstream	O
target	O
of	O
metabotropic	O
glutamate	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
Grm1	B-Gene_or_gene_product
)	O
.	O

We	O
reported	O
earlier	O
on	O
the	O
oncogenic	O
properties	O
of	O
Grm1	B-Gene_or_gene_product
by	O
demonstrating	O
that	O
stable	O
Grm1	B-Gene_or_gene_product
-	O
mouse	B-Organism
-	O
melanocytic	B-Cell
clones	I-Cell
proliferate	O
in	O
the	O
absence	O
of	O
growth	O
supplement	O
and	O
anchorage	O
in	O
vitro	O
.	O

In	O
addition	O
,	O
these	O
clones	B-Cell
also	O
exhibit	O
aggressive	O
tumorigenic	O
phenotypes	O
in	O
vivo	O
with	O
short	O
latency	O
in	O
tumor	B-Cancer
formation	O
in	O
both	O
immunodeficient	B-Organism
and	O
syngeneic	B-Organism
mice	I-Organism
.	O

We	O
also	O
detected	O
strong	O
activation	O
of	O
AKT	B-Gene_or_gene_product
in	O
allograft	B-Cancer
tumors	I-Cancer
specifically	O
AKT2	B-Gene_or_gene_product
as	O
the	O
predominant	O
isoform	O
involved	O
.	O

In	O
parallel	O
,	O
we	O
assessed	O
several	O
human	B-Organism
melanoma	B-Tissue
biopsy	I-Tissue
samples	I-Tissue
and	O
found	O
again	O
that	O
AKT2	B-Gene_or_gene_product
was	O
the	O
predominantly	O
activated	O
AKT	B-Gene_or_gene_product
in	O
these	O
human	B-Organism
melanoma	B-Tissue
biopsies	I-Tissue
.	O

In	O
cultured	O
stable	O
Grm1	B-Gene_or_gene_product
-	O
mouse	B-Organism
-	O
melanocytic	B-Cell
clones	I-Cell
,	O
as	O
well	O
as	O
an	O
metabotropic	O
glutamate	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
Grm1	B-Gene_or_gene_product
)	O
expressing	O
human	B-Organism
melanoma	B-Cell
cell	I-Cell
line	I-Cell
,	I-Cell
C8161	I-Cell
,	O
stimulation	O
of	O
Grm1	B-Gene_or_gene_product
by	O
its	O
agonist	O
led	O
to	O
the	O
activation	O
of	O
AKT	B-Gene_or_gene_product
,	O
while	O
preincubation	O
with	O
Grm1	B-Gene_or_gene_product
-	O
antagonist	O
abolished	O
Grm1	B-Gene_or_gene_product
-	O
agonist	O
-	O
induced	O
AKT	B-Gene_or_gene_product
activation	O
.	O

In	O
addition	O
,	O
a	O
reduction	O
in	O
tumor	B-Cancer
volume	O
of	O
Grm1	B-Gene_or_gene_product
-	O
mouse	B-Organism
-	O
melanocytic	B-Tissue
-	I-Tissue
allografts	I-Tissue
was	O
detected	O
in	O
the	O
presence	O
of	O
small	O
interfering	O
AKT2	B-Gene_or_gene_product
RNA	O
(	O
siAKT2	B-Gene_or_gene_product
)	O
.	O

Taken	O
together	O
,	O
these	O
results	O
showed	O
that	O
,	O
in	O
addition	O
to	O
the	O
MAPK	B-Gene_or_gene_product
pathway	O
previously	O
reported	O
being	O
a	O
downstream	O
target	O
of	O
stimulated	O
Grm1	B-Gene_or_gene_product
,	O
AKT2	B-Gene_or_gene_product
is	O
another	O
downstream	O
target	O
in	O
Grm1	B-Gene_or_gene_product
mediated	O
melanocyte	B-Cell
transformation	O
.	O

Hormonal	O
regulation	O
and	O
distinct	O
functions	O
of	O
semaphorin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3B	I-Gene_or_gene_product
and	O
semaphorin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3F	I-Gene_or_gene_product
in	O
ovarian	B-Cancer
cancer	I-Cancer
.	O

Semaphorins	B-Gene_or_gene_product
comprise	O
a	O
family	O
of	O
molecules	O
that	O
influence	O
neuronal	B-Cell
growth	O
and	O
guidance	O
.	O

Class	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
semaphorins	I-Gene_or_gene_product
,	O
semaphorin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3B	I-Gene_or_gene_product
(	O
SEMA3B	B-Gene_or_gene_product
)	O
and	O
semaphorin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3F	I-Gene_or_gene_product
(	O
SEMA3F	B-Gene_or_gene_product
)	O
,	O
illustrate	O
their	O
effects	O
by	O
forming	O
a	O
complex	O
with	O
neuropilins	B-Gene_or_gene_product
(	O
NP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
or	O
NP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
)	O
and	O
plexins	B-Gene_or_gene_product
.	O

We	O
examined	O
the	O
status	O
and	O
regulation	O
of	O
semaphorins	B-Gene_or_gene_product
and	O
their	O
receptors	O
in	O
human	B-Organism
ovarian	B-Cell
cancer	I-Cell
cells	I-Cell
.	O

A	O
significantly	O
reduced	O
expression	O
of	O
SEMA3B	B-Gene_or_gene_product
(	O
83	O
kDa	O
)	O
,	O
SEMA3F	B-Gene_or_gene_product
(	O
90	O
kDa	O
)	O
,	O
and	O
plexin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
A3	I-Gene_or_gene_product
was	O
observed	O
in	O
ovarian	B-Cell
cancer	I-Cell
cell	I-Cell
lines	I-Cell
when	O
compared	O
with	O
normal	O
human	B-Organism
ovarian	B-Cell
surface	I-Cell
epithelial	I-Cell
cells	I-Cell
.	O

The	O
expression	O
of	O
NP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
NP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
and	O
plexin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
A1	I-Gene_or_gene_product
was	O
not	O
altered	O
in	O
human	B-Organism
ovarian	B-Cell
surface	I-Cell
epithelial	I-Cell
and	O
ovarian	B-Cell
cancer	I-Cell
cells	I-Cell
.	O

The	O
decreased	O
expression	O
of	O
SEMA3B	B-Gene_or_gene_product
,	O
SEMA3F	B-Gene_or_gene_product
,	O
and	O
plexin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
A3	I-Gene_or_gene_product
was	O
confirmed	O
in	O
stage	O
3	O
ovarian	B-Cancer
tumors	I-Cancer
.	O

The	O
treatment	O
of	O
ovarian	B-Cell
cancer	I-Cell
cells	I-Cell
with	O
luteinizing	B-Gene_or_gene_product
hormone	I-Gene_or_gene_product
,	O
follicle	B-Gene_or_gene_product
-	I-Gene_or_gene_product
stimulating	I-Gene_or_gene_product
hormone	I-Gene_or_gene_product
,	O
and	O
estrogen	B-Simple_chemical
induced	O
a	O
significant	O
upregulation	O
of	O
SEMA3B	B-Gene_or_gene_product
,	O
whereas	O
SEMA3F	B-Gene_or_gene_product
was	O
upregulated	O
only	O
by	O
estrogen	B-Simple_chemical
.	O

Cotreatment	O
of	O
cell	B-Cell
lines	I-Cell
with	O
a	O
hormone	O
and	O
its	O
specific	O
antagonist	O
blocked	O
the	O
effect	O
of	O
the	O
hormone	O
.	O

Ectopic	O
expression	O
of	O
SEMA3B	B-Gene_or_gene_product
or	O
SEMA3F	B-Gene_or_gene_product
reduced	O
soft	O
-	O
agar	O
colony	O
formation	O
,	O
adhesion	O
,	O
and	O
cell	B-Cell
invasion	O
of	O
ovarian	B-Cell
cancer	I-Cell
cell	I-Cell
cultures	I-Cell
.	O

Forced	O
expression	O
of	O
SEMA3B	B-Gene_or_gene_product
,	O
but	O
not	O
SEMA3F	B-Gene_or_gene_product
,	O
inhibited	O
viability	O
of	O
ovarian	B-Cell
cancer	I-Cell
cells	I-Cell
.	O

Overexpression	O
of	O
SEMA3B	B-Gene_or_gene_product
and	O
SEMA3F	B-Gene_or_gene_product
reduced	O
focal	B-Gene_or_gene_product
adhesion	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
phosphorylation	O
and	O
matrix	B-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
matrix	B-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
expression	O
in	O
ovarian	B-Cell
cancer	I-Cell
cells	I-Cell
.	O

Forced	O
expression	O
of	O
SEMA3F	B-Gene_or_gene_product
,	O
but	O
not	O
SEMA3B	B-Gene_or_gene_product
in	O
ovarian	B-Cell
cancer	I-Cell
cells	I-Cell
,	O
significantly	O
inhibited	O
endothelial	B-Tissue
cell	I-Tissue
tube	I-Tissue
formation	O
.	O

Collectively	O
,	O
our	O
results	O
suggest	O
that	O
the	O
loss	O
of	O
SEMA3	B-Gene_or_gene_product
expression	O
could	O
be	O
a	O
hallmark	O
of	O
cancer	B-Cancer
progression	O
.	O

Furthermore	O
,	O
gonadotropin	B-Gene_or_gene_product
-	O
and	O
/	O
or	O
estrogen	B-Simple_chemical
-	O
mediated	O
maintenance	O
of	O
SEMA3	B-Gene_or_gene_product
expression	O
could	O
control	O
ovarian	B-Cancer
cancer	I-Cancer
angiogenesis	O
and	O
metastasis	O
.	O

Tobacco	B-Organism
smoke	O
promotes	O
lung	B-Organ
tumorigenesis	O
by	O
triggering	O
IKKbeta	B-Gene_or_gene_product
-	O
and	O
JNK1	B-Gene_or_gene_product
-	O
dependent	O
inflammation	B-Pathological_formation
.	O

Chronic	O
exposure	O
to	O
tobacco	B-Organism
smoke	O
,	O
which	O
contains	O
over	O
60	O
tumor	B-Cancer
-	O
initiating	O
carcinogens	O
,	O
is	O
the	O
major	O
risk	O
factor	O
for	O
development	O
of	O
lung	B-Cancer
cancer	I-Cancer
,	O
accounting	O
for	O
a	O
large	O
portion	O
of	O
cancer	B-Cancer
-	O
related	O
deaths	O
worldwide	O
.	O

It	O
is	O
well	O
established	O
that	O
tobacco	B-Organism
smoke	O
is	O
a	O
tumor	B-Cancer
initiator	O
,	O
but	O
we	O
asked	O
whether	O
it	O
also	O
acts	O
as	O
a	O
tumor	B-Cancer
promoter	O
once	O
malignant	O
initiation	O
,	O
such	O
as	O
caused	O
by	O
K	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
activation	O
,	O
has	O
taken	O
place	O
.	O

Here	O
we	O
demonstrate	O
that	O
repetitive	O
exposure	O
to	O
tobacco	B-Organism
smoke	O
promotes	O
tumor	B-Cancer
development	O
both	O
in	O
carcinogen	O
-	O
treated	O
mice	B-Organism
and	O
in	O
transgenic	O
mice	B-Organism
undergoing	O
sporadic	O
K	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
activation	O
in	O
lung	B-Cell
epithelial	I-Cell
cells	I-Cell
.	O

Tumor	B-Cancer
promotion	O
is	O
due	O
to	O
induction	O
of	O
inflammation	B-Pathological_formation
that	O
results	O
in	O
enhanced	O
pneumocyte	B-Cell
proliferation	O
and	O
is	O
abrogated	O
by	O
IKKbeta	B-Gene_or_gene_product
ablation	O
in	O
myeloid	B-Cell
cells	I-Cell
or	O
inactivation	O
of	O
JNK1	B-Gene_or_gene_product
.	O

Dose	O
-	O
and	O
time	O
-	O
dependent	O
effects	O
of	O
doxorubicin	B-Simple_chemical
on	O
cytotoxicity	O
,	O
cell	B-Cell
cycle	O
and	O
apoptotic	O
cell	B-Cell
death	O
in	O
human	B-Organism
colon	B-Cell
cancer	I-Cell
cells	I-Cell
.	O

The	O
cytostatic	O
drug	O
doxorubicin	B-Simple_chemical
is	O
a	O
well	O
-	O
known	O
chemotherapeutic	O
agent	O
which	O
is	O
used	O
in	O
treatment	O
of	O
a	O
wide	O
variety	O
of	O
cancers	B-Cancer
.	O

A	O
key	O
factor	O
in	O
the	O
response	O
of	O
cancer	B-Cell
cells	I-Cell
to	O
chemotherapeutic	O
drugs	O
is	O
the	O
activation	O
of	O
the	O
apoptotic	O
pathway	O
,	O
a	O
pathway	O
that	O
is	O
often	O
impaired	O
in	O
chemoresistant	O
colon	B-Cell
cancer	I-Cell
cells	I-Cell
.	O

The	O
aim	O
of	O
the	O
present	O
study	O
was	O
to	O
investigate	O
the	O
effects	O
of	O
doxorubicin	B-Simple_chemical
in	O
Hct	B-Cell
-	I-Cell
116	I-Cell
human	I-Cell
colon	I-Cell
carcinoma	I-Cell
cells	I-Cell
in	O
order	O
to	O
clarify	O
if	O
a	O
time	O
/	O
concentration	O
range	O
for	O
optimal	O
doxorubicin	B-Simple_chemical
-	O
induced	O
apoptosis	O
exists	O
.	O

We	O
compared	O
a	O
treatment	O
schedule	O
were	O
cells	B-Cell
were	O
bolus	O
incubated	O
for	O
3h	O
with	O
doxorubicin	B-Simple_chemical
followed	O
by	O
24h	O
in	O
drug	O
-	O
free	O
medium	O
,	O
with	O
a	O
continuous	O
doxorubicin	B-Simple_chemical
treatment	O
schedule	O
for	O
24h	O
.	O

Bolus	O
incubation	O
was	O
carried	O
out	O
to	O
determine	O
effects	O
of	O
doxorubicin	B-Simple_chemical
accumulated	O
during	O
the	O
first	O
3h	O
,	O
whereas	O
continuous	O
incubation	O
allowed	O
further	O
(	O
continuous	O
)	O
exposure	O
to	O
doxorubicin	B-Simple_chemical
.	O

We	O
found	O
that	O
bolus	O
(	O
3h	O
)	O
treatment	O
with	O
doxorubicin	B-Simple_chemical
resulted	O
in	O
a	O
dose	O
-	O
dependent	O
decrease	O
of	O
viable	O
cells	B-Cell
and	O
concomitant	O
increase	O
of	O
apoptosis	O
.	O

Additionally	O
,	O
bolus	O
(	O
3h	O
)	O
doxorubicin	B-Simple_chemical
incubation	O
led	O
to	O
phosphorylation	O
of	O
p53	B-Gene_or_gene_product
at	O
serine	B-Amino_acid
392	I-Amino_acid
,	O
induction	O
of	O
p21	B-Gene_or_gene_product
,	O
G2	O
arrest	O
and	O
increase	O
of	O
proapoptotic	O
protein	O
Bax	B-Gene_or_gene_product
.	O

In	O
contrast	O
,	O
continuous	O
(	O
24h	O
)	O
treatment	O
with	O
doxorubicin	B-Simple_chemical
reduced	O
the	O
number	O
of	O
living	O
cells	B-Cell
with	O
no	O
parallel	O
raise	O
in	O
the	O
amount	O
of	O
dead	O
cells	B-Cell
.	O

Continuous	O
(	O
24h	O
)	O
treatment	O
with	O
5	O
microM	O
doxorubicin	B-Simple_chemical
resulted	O
in	O
cell	B-Cell
cycle	O
arrest	O
in	O
G0	O
/	O
G1	O
phase	O
that	O
was	O
neither	O
accompanied	O
by	O
phosphorylation	O
and	O
activation	O
of	O
p53	B-Gene_or_gene_product
nor	O
enhanced	O
expression	O
of	O
p21	B-Gene_or_gene_product
.	O

These	O
results	O
suggest	O
that	O
doxorubicin	B-Simple_chemical
is	O
able	O
to	O
induce	O
cell	B-Cell
death	O
by	O
apoptosis	O
only	O
at	O
particular	O
dose	O
and	O
treatment	O
conditions	O
and	O
imply	O
a	O
completely	O
different	O
cellular	B-Cell
response	O
following	O
bolus	O
or	O
continuous	O
exposure	O
to	O
doxorubicin	B-Simple_chemical
.	O

Lactate	B-Gene_or_gene_product
-	I-Gene_or_gene_product
dehydrogenase	I-Gene_or_gene_product
5	I-Gene_or_gene_product
is	O
overexpressed	O
in	O
non	B-Cancer
-	I-Cancer
small	I-Cancer
cell	I-Cancer
lung	I-Cancer
cancer	I-Cancer
and	O
correlates	O
with	O
the	O
expression	O
of	O
the	O
transketolase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
like	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

AIMS	O
:	O
As	O
one	O
of	O
the	O
five	O
lactate	B-Gene_or_gene_product
dehydrogenase	I-Gene_or_gene_product
(	O
LDH	B-Gene_or_gene_product
)	O
isoenzymes	O
,	O
LDH5	B-Gene_or_gene_product
has	O
the	O
highest	O
efficiency	O
to	O
catalyze	O
pyruvate	B-Simple_chemical
transformation	O
to	O
lactate	B-Simple_chemical
.	O

LDH5	B-Gene_or_gene_product
overexpression	O
in	O
cancer	B-Cell
cells	I-Cell
induces	O
an	O
upregulated	O
glycolytic	O
metabolism	O
and	O
reduced	O
dependence	O
on	O
the	O
presence	O
of	O
oxygen	B-Simple_chemical
.	O

Here	O
we	O
analyzed	O
LDH5	B-Gene_or_gene_product
protein	O
expression	O
in	O
a	O
well	O
characterized	O
large	O
cohort	O
of	O
primary	B-Cancer
lung	I-Cancer
cancers	I-Cancer
in	O
correlation	O
to	O
clinico	O
-	O
pathological	O
data	O
and	O
its	O
possible	O
impact	O
on	O
patient	B-Organism
survival	O
.	O

METHODS	O
:	O
Primary	B-Cancer
lung	I-Cancer
cancers	I-Cancer
(	O
n	O
=	O
269	O
)	O
and	O
non	B-Tissue
neoplastic	I-Tissue
lung	I-Tissue
tissue	I-Tissue
(	O
n	O
=	O
35	O
)	O
were	O
tested	O
for	O
LDH5	B-Gene_or_gene_product
expression	O
by	O
immunohistochemistry	O
using	O
a	O
polyclonal	O
LDH5	B-Gene_or_gene_product
antibody	O
(	O
ab53010	O
)	O
.	O

The	O
results	O
of	O
LDH5	B-Gene_or_gene_product
expression	O
were	O
correlated	O
to	O
clinico	O
-	O
pathological	O
data	O
as	O
well	O
as	O
to	O
patient	B-Organism
'	O
s	O
survival	O
.	O

In	O
addition	O
,	O
the	O
results	O
of	O
the	O
previously	O
tested	O
transketolase	B-Gene_or_gene_product
like	I-Gene_or_gene_product
1	I-Gene_or_gene_product
protein	O
(	O
TKTL1	B-Gene_or_gene_product
)	O
expression	O
were	O
correlated	O
to	O
LDH5	B-Gene_or_gene_product
expression	O
.	O

RESULTS	O
:	O
89	O
.	O
5	O
%	O
(	O
n	O
=	O
238	O
)	O
of	O
NSCLC	B-Cancer
revealed	O
LDH5	B-Gene_or_gene_product
expression	O
whereas	O
LDH5	B-Gene_or_gene_product
expression	O
was	O
not	O
detected	O
in	O
non	B-Tissue
neoplastic	I-Tissue
lung	I-Tissue
tissues	I-Tissue
(	O
n	O
=	O
34	O
)	O
(	O
p	O
<	O
0	O
.	O
0001	O
)	O
.	O

LDH5	B-Gene_or_gene_product
overexpression	O
was	O
associated	O
with	O
histological	O
type	O
(	O
adenocarcinoma	B-Cancer
=	O
57	O
%	O
,	O
squamous	B-Cancer
cell	I-Cancer
carcinoma	I-Cancer
=	O
45	O
%	O
,	O
large	B-Cancer
cell	I-Cancer
carcinoma	I-Cancer
=	O
46	O
%	O
,	O
p	O
=	O
0	O
.	O
006	O
)	O
.	O

No	O
significant	O
correlation	O
could	O
be	O
detected	O
with	O
regard	O
to	O
TNM	O
-	O
stage	O
,	O
grading	O
or	O
survival	O
.	O

A	O
two	O
sided	O
correlation	O
between	O
the	O
expression	O
of	O
TKTL1	B-Gene_or_gene_product
and	O
LDH5	B-Gene_or_gene_product
could	O
be	O
shown	O
(	O
p	O
=	O
0	O
.	O
002	O
)	O
within	O
the	O
overall	O
cohort	O
as	O
well	O
as	O
for	O
each	O
grading	O
and	O
pN	O
group	O
.	O

A	O
significant	O
correlation	O
between	O
LDH5	B-Gene_or_gene_product
and	O
TKTL1	B-Gene_or_gene_product
within	O
each	O
histologic	O
tumortype	B-Cancer
could	O
not	O
be	O
revealed	O
.	O

CONCLUSIONS	O
:	O
LDH5	B-Gene_or_gene_product
is	O
overexpressed	O
in	O
NSCLC	B-Cancer
and	O
could	O
hence	O
serve	O
as	O
an	O
additional	O
marker	O
for	O
malignancy	O
.	O

Furthermore	O
,	O
LDH5	B-Gene_or_gene_product
correlates	O
positively	O
with	O
the	O
prognostic	O
marker	O
TKTL1	B-Gene_or_gene_product
.	O

Our	O
results	O
confirm	O
a	O
close	O
link	O
between	O
the	O
two	O
metabolic	O
enzymes	O
and	O
indicate	O
an	O
alteration	O
in	O
the	O
glucose	B-Simple_chemical
metabolism	O
in	O
the	O
process	O
of	O
malignant	O
transformation	O
.	O

Telomere	B-Cellular_component
/	O
telomerase	B-Gene_or_gene_product
interplay	O
in	O
virus	O
-	O
driven	O
and	O
virus	O
-	O
independent	O
lymphomagenesis	O
:	O
pathogenic	O
and	O
clinical	O
implications	O
.	O

Telomerase	B-Gene_or_gene_product
is	O
a	O
ribonucleoprotein	O
complex	O
critically	O
involved	O
in	O
extending	O
and	O
maintaining	O
telomeres	B-Cellular_component
.	O

Unlike	O
the	O
majority	O
of	O
somatic	B-Cell
cells	I-Cell
,	O
in	O
which	O
hTERT	B-Gene_or_gene_product
and	O
telomerase	B-Gene_or_gene_product
activity	O
are	O
generally	O
silent	O
,	O
normal	O
lymphocytes	B-Cell
show	O
transient	O
physiological	O
hTERT	B-Gene_or_gene_product
expression	O
and	O
telomerase	B-Gene_or_gene_product
activity	O
according	O
to	O
their	O
differentiation	O
/	O
activation	O
status	O
.	O

During	O
lymphomagenesis	O
,	O
induction	O
of	O
persistent	O
telomerase	B-Gene_or_gene_product
expression	O
and	O
activity	O
may	O
occur	O
before	O
or	O
after	O
telomere	B-Cellular_component
shortening	O
,	O
as	O
a	O
consequence	O
of	O
the	O
different	O
mechanisms	O
through	O
which	O
transforming	O
factors	O
/	O
agents	O
may	O
activate	O
telomerase	B-Gene_or_gene_product
.	O

Available	O
data	O
indicate	O
that	O
the	O
timing	O
of	O
telomerase	B-Gene_or_gene_product
activation	O
may	O
allow	O
the	O
distinction	O
of	O
two	O
different	O
lymphomagenetic	O
models	O
:	O
(	O
i	O
)	O
an	O
early	O
activation	O
of	O
telomerase	B-Gene_or_gene_product
via	O
exogenous	O
regulators	O
of	O
hTERT	B-Gene_or_gene_product
,	O
along	O
with	O
an	O
increased	O
lymphocyte	B-Cell
growth	O
and	O
a	O
subsequent	O
selection	O
of	O
cells	B-Cell
with	O
increased	O
transforming	O
potential	O
may	O
characterize	O
several	O
virus	O
-	O
related	O
lymphoid	B-Pathological_formation
malignancies	I-Pathological_formation
;	O
(	O
ii	O
)	O
a	O
progressive	O
shortening	O
of	O
telomeres	B-Cellular_component
,	O
leading	O
to	O
genetic	O
instability	O
which	O
favors	O
a	O
subsequent	O
activation	O
of	O
telomerase	B-Gene_or_gene_product
via	O
endogenous	O
regulators	O
may	O
occur	O
in	O
most	O
virus	O
-	O
unrelated	O
lymphoid	B-Cancer
tumors	I-Cancer
.	O

These	O
models	O
may	O
have	O
clinically	O
relevant	O
implications	O
,	O
particularly	O
for	O
the	O
tailoring	O
of	O
therapeutic	O
strategies	O
targeting	O
telomerase	B-Gene_or_gene_product
.	O

Betulin	B-Simple_chemical
induces	O
mitochondrial	B-Cellular_component
cytochrome	B-Gene_or_gene_product
c	I-Gene_or_gene_product
release	O
associated	O
apoptosis	O
in	O
human	B-Organism
cancer	B-Cell
cells	I-Cell
.	O

We	O
examined	O
whether	O
betulin	B-Simple_chemical
,	O
a	O
naturally	O
abundant	O
compound	O
,	O
has	O
anticancer	B-Cancer
functions	O
in	O
human	B-Organism
cancer	B-Cell
cells	I-Cell
.	O

The	O
results	O
showed	O
that	O
betulin	B-Simple_chemical
significantly	O
inhibited	O
cell	B-Cell
viability	O
in	O
cervix	B-Cell
carcinoma	I-Cell
HeLa	I-Cell
cells	I-Cell
,	O
hepatoma	B-Cell
HepG2	I-Cell
cells	I-Cell
,	O
lung	B-Cell
adenocarcinoma	I-Cell
A549	I-Cell
cells	I-Cell
,	O
and	O
breast	B-Cell
cancer	I-Cell
MCF	I-Cell
-	I-Cell
7	I-Cell
cells	I-Cell
with	O
IC	O
(	O
50	O
)	O
values	O
ranging	O
from	O
10	O
to	O
15	O
microg	O
/	O
mL	O
.	O

While	O
betulin	B-Simple_chemical
exhibited	O
only	O
moderate	O
anticancer	B-Cancer
activity	O
in	O
other	O
human	B-Organism
cancer	B-Cell
cells	I-Cell
such	O
as	O
hepatoma	B-Cell
SK	I-Cell
-	I-Cell
HEP	I-Cell
-	I-Cell
1	I-Cell
cells	I-Cell
,	O
prostate	B-Cell
carcinoma	I-Cell
PC	I-Cell
-	I-Cell
3	I-Cell
,	O
and	O
lung	B-Cell
carcinoma	I-Cell
NCI	I-Cell
-	I-Cell
H460	I-Cell
,	O
with	O
IC	O
(	O
50	O
)	O
values	O
ranging	O
from	O
20	O
to	O
60	O
microg	O
/	O
mL	O
,	O
it	O
showed	O
minor	O
growth	O
inhibition	O
in	O
human	B-Organism
erythroleukemia	B-Cell
K562	I-Cell
cells	I-Cell
(	O
IC	O
(	O
50	O
)	O
>	O
100	O
microg	O
/	O
mL	O
)	O
.	O

We	O
further	O
investigated	O
the	O
mechanism	O
of	O
anticancer	B-Cancer
activity	O
by	O
betulin	B-Simple_chemical
,	O
using	O
HeLa	B-Cell
cells	I-Cell
as	O
an	O
experimental	O
model	O
.	O

Betulin	B-Simple_chemical
(	O
10	O
microg	O
/	O
mL	O
)	O
induces	O
apoptotic	O
cell	B-Cell
death	O
,	O
as	O
evidenced	O
by	O
morphological	O
characteristics	O
such	O
as	O
membrane	B-Cellular_component
phosphatidylserine	B-Amino_acid
translocation	O
,	O
nuclear	B-Cellular_component
condensation	O
/	O
fragmentation	O
,	O
and	O
apoptotic	B-Cellular_component
body	I-Cellular_component
formation	O
.	O

A	O
kinetics	O
analysis	O
showed	O
that	O
the	O
depolarization	O
of	O
mitochondrial	B-Cellular_component
membrane	I-Cellular_component
potential	O
and	O
the	O
release	O
of	O
mitochondrial	B-Cellular_component
cytochrome	B-Gene_or_gene_product
c	I-Gene_or_gene_product
occurred	O
as	O
early	O
as	O
30	O
min	O
after	O
treatment	O
with	O
betulin	B-Simple_chemical
.	O

Betulin	B-Simple_chemical
,	O
unlike	O
its	O
chemical	O
derivative	O
betulinic	B-Simple_chemical
acid	I-Simple_chemical
,	O
did	O
not	O
directly	O
trigger	O
mitochondrial	B-Cellular_component
cytochrome	B-Gene_or_gene_product
c	I-Gene_or_gene_product
release	O
in	O
isolated	O
mitochondria	B-Cellular_component
.	O

Importantly	O
,	O
Bax	B-Gene_or_gene_product
and	O
Bak	B-Gene_or_gene_product
were	O
rapidly	O
translocated	O
to	O
the	O
mitochondria	B-Cellular_component
30	O
min	O
after	O
betulin	B-Simple_chemical
treatment	O
.	O

The	O
sequential	O
activation	O
of	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
and	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
/	O
-	B-Gene_or_gene_product
7	I-Gene_or_gene_product
and	O
the	O
cleavage	O
of	O
poly	B-Gene_or_gene_product
(	I-Gene_or_gene_product
ADP	I-Gene_or_gene_product
-	I-Gene_or_gene_product
ribose	I-Gene_or_gene_product
)	I-Gene_or_gene_product
polymerase	I-Gene_or_gene_product
(	O
PARP	B-Gene_or_gene_product
)	O
were	O
observed	O
behind	O
those	O
mitochondrial	B-Cellular_component
events	O
.	O

Furthermore	O
,	O
specific	O
downregulation	O
of	O
either	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
,	O
Bax	B-Gene_or_gene_product
,	O
or	O
Bak	B-Gene_or_gene_product
by	O
siRNA	O
effectively	O
reduced	O
PARP	B-Gene_or_gene_product
cleavage	O
and	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
activation	O
.	O

Taken	O
together	O
,	O
the	O
lines	O
of	O
evidence	O
demonstrate	O
that	O
betulin	B-Simple_chemical
triggers	O
apoptosis	O
of	O
human	B-Organism
cancer	B-Cell
cells	I-Cell
through	O
the	O
intrinsic	O
apoptotic	O
pathway	O
.	O

Adenovirus	B-Organism
5	I-Organism
E1A	B-Gene_or_gene_product
enhances	O
histone	B-Gene_or_gene_product
deacetylase	I-Gene_or_gene_product
inhibitors	O
-	O
induced	O
apoptosis	O
through	O
Egr	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
mediated	O
Bim	B-Gene_or_gene_product
upregulation	O
.	O

Histone	B-Gene_or_gene_product
deacetylase	I-Gene_or_gene_product
inhibitors	O
(	O
HDACi	O
)	O
are	O
potent	O
anti	O
-	O
cancer	B-Cancer
agents	O
for	O
variety	O
of	O
cancer	B-Cancer
types	O
.	O

Suberoylanilide	B-Simple_chemical
hydroxamic	I-Simple_chemical
acid	I-Simple_chemical
(	O
SAHA	B-Simple_chemical
)	O
has	O
been	O
approved	O
as	O
a	O
drug	O
to	O
treat	O
cutaneous	O
T	B-Cancer
cell	I-Cancer
lymphoma	I-Cancer
,	O
and	O
the	O
combination	O
of	O
HDACi	O
and	O
other	O
agents	O
have	O
been	O
actively	O
tested	O
in	O
many	O
clinical	O
trials	O
.	O

Adenovirus	B-Organism
5	I-Organism
early	B-Gene_or_gene_product
region	I-Gene_or_gene_product
1A	I-Gene_or_gene_product
(	O
E1A	B-Gene_or_gene_product
)	O
has	O
been	O
shown	O
to	O
exhibit	O
high	O
tumor	B-Cancer
suppressor	O
activity	O
,	O
and	O
gene	O
therapy	O
using	O
E1A	B-Gene_or_gene_product
has	O
been	O
tested	O
in	O
clinical	O
trials	O
.	O

Here	O
,	O
we	O
showed	O
that	O
proapoptotic	O
activity	O
of	O
HDACi	O
was	O
robustly	O
enhanced	O
by	O
E1A	B-Gene_or_gene_product
in	O
multiple	B-Cell
cancer	I-Cell
cells	I-Cell
,	O
but	O
not	O
in	O
normal	O
cells	B-Cell
.	O

Moreover	O
,	O
we	O
showed	O
that	O
combination	O
of	O
E1A	B-Gene_or_gene_product
gene	O
therapy	O
and	O
SAHA	B-Simple_chemical
showed	O
high	O
therapeutic	O
efficacy	O
with	O
low	O
toxicity	O
in	O
vivo	O
ovarian	B-Cancer
and	O
breast	B-Cancer
xenograft	I-Cancer
models	O
.	O

SAHA	B-Simple_chemical
downregulated	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
XL	I-Gene_or_gene_product
and	O
upregulated	O
proapoptotic	O
BH3	O
-	O
only	O
protein	O
Bim	B-Gene_or_gene_product
,	O
whose	O
expression	O
was	O
further	O
enhanced	O
by	O
E1A	B-Gene_or_gene_product
in	O
cancer	B-Cell
cells	I-Cell
.	O

These	O
alterations	O
of	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
family	O
proteins	O
were	O
critical	O
for	O
apoptosis	O
induced	O
by	O
the	O
combination	O
in	O
cancer	B-Cell
cells	I-Cell
.	O

SAHA	B-Simple_chemical
enhanced	O
acetylation	O
of	O
histone	B-Gene_or_gene_product
H3	I-Gene_or_gene_product
in	O
Bim	B-Gene_or_gene_product
promoter	O
region	O
,	O
while	O
E1A	B-Gene_or_gene_product
upregulated	O
Egr	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
which	O
was	O
directly	O
involved	O
in	O
Bim	B-Gene_or_gene_product
transactivation	O
.	O

Together	O
,	O
our	O
results	O
provide	O
not	O
only	O
a	O
novel	O
insight	O
into	O
the	O
mechanisms	O
underlying	O
anti	O
-	O
tumor	B-Cancer
activity	O
of	O
E1A	B-Gene_or_gene_product
,	O
but	O
also	O
a	O
rationale	O
for	O
the	O
combined	O
HDACi	O
and	O
E1A	B-Gene_or_gene_product
gene	O
therapy	O
in	O
future	O
clinical	O
trials	O
.	O

Constitutively	O
nuclear	B-Cellular_component
FOXO3a	B-Gene_or_gene_product
localization	O
predicts	O
poor	O
survival	O
and	O
promotes	O
Akt	B-Gene_or_gene_product
phosphorylation	O
in	O
breast	B-Cancer
cancer	I-Cancer
.	O

BACKGROUND	O
:	O
The	O
PI3K	B-Gene_or_gene_product
-	O
Akt	B-Gene_or_gene_product
signal	O
pathway	O
plays	O
a	O
key	O
role	O
in	O
tumorigenesis	O
and	O
the	O
development	O
of	O
drug	O
-	O
resistance	O
.	O

Cytotoxic	O
chemotherapy	O
resistance	O
is	O
linked	O
to	O
limited	O
therapeutic	O
options	O
and	O
poor	O
prognosis	O
.	O

METHODOLOGY	O
/	O
PRINCIPAL	O
FINDINGS	O
:	O
Examination	O
of	O
FOXO3a	B-Gene_or_gene_product
and	O
phosphorylated	O
-	O
Akt	B-Gene_or_gene_product
(	O
P	O
-	O
Akt	B-Gene_or_gene_product
)	O
expression	O
in	O
breast	B-Cancer
cancer	I-Cancer
tissue	I-Cancer
microarrays	O
showed	O
nuclear	B-Cellular_component
FOXO3a	B-Gene_or_gene_product
was	O
associated	O
with	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
positivity	O
(	O
p	O
=	O
0	O
.	O
052	O
)	O
,	O
poor	O
prognosis	O
(	O
p	O
=	O
0	O
.	O
014	O
)	O
,	O
and	O
P	O
-	O
Akt	B-Gene_or_gene_product
expression	O
in	O
invasive	B-Cancer
ductal	I-Cancer
carcinoma	I-Cancer
.	O

Using	O
tamoxifen	B-Simple_chemical
and	O
doxorubicin	B-Simple_chemical
-	O
sensitive	O
and	O
-	O
resistant	O
breast	B-Cell
cancer	I-Cell
cell	I-Cell
lines	I-Cell
as	O
models	O
,	O
we	O
found	O
that	O
doxorubicin	B-Simple_chemical
-	O
but	O
not	O
tamoxifen	B-Simple_chemical
-	O
resistance	O
is	O
associated	O
with	O
nuclear	B-Cellular_component
accumulation	O
of	O
FOXO3a	B-Gene_or_gene_product
,	O
consistent	O
with	O
the	O
finding	O
that	O
sustained	O
nuclear	B-Cellular_component
FOXO3a	B-Gene_or_gene_product
is	O
associated	O
with	O
poor	O
prognosis	O
.	O

We	O
also	O
established	O
that	O
doxorubicin	B-Simple_chemical
treatment	O
induces	O
proliferation	O
arrest	O
and	O
FOXO3a	B-Gene_or_gene_product
nuclear	B-Cellular_component
relocation	O
in	O
sensitive	O
breast	B-Cell
cancer	I-Cell
cells	I-Cell
.	O

Induction	O
of	O
FOXO3a	B-Gene_or_gene_product
activity	O
in	O
doxorubicin	B-Simple_chemical
-	O
sensitive	O
MCF	B-Cell
-	I-Cell
7	I-Cell
cells	I-Cell
was	O
sufficient	O
to	O
promote	O
Akt	B-Gene_or_gene_product
phosphorylation	O
and	O
arrest	O
cell	B-Cell
proliferation	O
.	O

Conversely	O
,	O
knockdown	O
of	O
endogenous	O
FOXO3a	B-Gene_or_gene_product
expression	O
reduced	O
PI3K	B-Gene_or_gene_product
/	O
Akt	B-Gene_or_gene_product
activity	O
.	O

Using	O
MDA	B-Cell
-	I-Cell
MB	I-Cell
-	I-Cell
231	I-Cell
cells	I-Cell
,	O
in	O
which	O
FOXO3a	B-Gene_or_gene_product
activity	O
can	O
be	O
induced	O
by	O
4	B-Simple_chemical
-	I-Simple_chemical
hydroxytamoxifen	I-Simple_chemical
,	O
we	O
showed	O
that	O
FOXO3a	B-Gene_or_gene_product
induction	O
up	O
-	O
regulates	O
PI3K	B-Gene_or_gene_product
-	O
Akt	B-Gene_or_gene_product
activity	O
and	O
enhanced	O
doxorubicin	B-Simple_chemical
resistance	O
.	O

However	O
FOXO3a	B-Gene_or_gene_product
induction	O
has	O
little	O
effect	O
on	O
cell	B-Cell
proliferation	O
,	O
indicating	O
that	O
FOXO3a	B-Gene_or_gene_product
or	O
its	O
downstream	O
activity	O
is	O
deregulated	O
in	O
the	O
cytotoxic	O
drug	O
resistant	O
breast	B-Cell
cancer	I-Cell
cells	I-Cell
.	O

Thus	O
,	O
our	O
results	O
suggest	O
that	O
sustained	O
FOXO3a	B-Gene_or_gene_product
activation	O
can	O
enhance	O
hyperactivation	O
of	O
the	O
PI3K	B-Gene_or_gene_product
/	O
Akt	B-Gene_or_gene_product
pathway	O
.	O

CONCLUSIONS	O
/	O
SIGNIFICANCE	O
:	O
Together	O
these	O
data	O
suggest	O
that	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
and	O
poor	O
survival	O
in	O
invasive	B-Cancer
ductal	I-Cancer
breast	I-Cancer
carcinoma	I-Cancer
are	O
linked	O
to	O
an	O
uncoupling	O
of	O
the	O
Akt	B-Gene_or_gene_product
-	O
FOXO3a	B-Gene_or_gene_product
signaling	O
axis	O
.	O

In	O
these	O
breast	B-Cancer
cancers	I-Cancer
activated	O
Akt	B-Gene_or_gene_product
fails	O
to	O
inactivate	O
and	O
re	O
-	O
localize	O
FOXO3a	B-Gene_or_gene_product
to	O
the	O
cytoplasm	B-Organism_substance
,	O
and	O
nuclear	B-Cellular_component
-	O
targeted	O
FOXO3a	B-Gene_or_gene_product
does	O
not	O
induce	O
cell	B-Cell
death	O
or	O
cell	B-Cell
cycle	O
arrest	O
.	O

As	O
such	O
,	O
sustained	O
nuclear	B-Cellular_component
FOXO3a	B-Gene_or_gene_product
expression	O
in	O
breast	B-Cancer
cancer	I-Cancer
may	O
culminate	O
in	O
cancer	B-Cancer
progression	O
and	O
the	O
development	O
of	O
an	O
aggressive	O
phenotype	O
similar	O
to	O
that	O
observed	O
in	O
cytotoxic	O
chemotherapy	O
resistant	O
breast	B-Cell
cancer	I-Cell
cell	I-Cell
models	O
.	O

Skin	B-Organ
deep	O
and	O
deeper	O
:	O
multiple	O
pathways	O
in	O
basal	B-Cell
cell	I-Cell
carcinogenesis	O
.	O

This	O
perspective	O
places	O
the	O
report	O
by	O
Villani	O
et	O
al	O
.	O
that	O
appears	O
in	O
this	O
issue	O
of	O
the	O
journal	O
(	O
beginning	O
on	O
page	O
1222	O
)	O
in	O
the	O
context	O
of	O
recent	O
work	O
showing	O
an	O
intersection	O
between	O
two	O
important	O
developmental	O
pathways	O
implicated	O
in	O
oncogenesis	O
:	O
the	O
hedgehog	B-Gene_or_gene_product
and	O
insulin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
like	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
IGF	B-Gene_or_gene_product
)	O
pathways	O
.	O

Villani	O
et	O
al	O
.	O
define	O
a	O
key	O
role	O
for	O
the	O
IGF	B-Gene_or_gene_product
regulatory	O
protein	O
Igfbp2	B-Gene_or_gene_product
in	O
a	O
genetic	O
model	O
of	O
basal	B-Cell
cell	I-Cell
carcinogenesis	O
driven	O
by	O
targeted	O
constitutive	O
activation	O
of	O
hedgehog	B-Gene_or_gene_product
signaling	O
.	O

Placed	O
in	O
the	O
framework	O
of	O
other	O
recently	O
published	O
work	O
,	O
the	O
observations	O
of	O
Villani	O
et	O
al	O
.	O
both	O
raise	O
questions	O
about	O
the	O
cell	B-Cell
of	O
origin	O
for	O
basal	B-Cancer
cell	I-Cancer
cancers	I-Cancer
and	O
define	O
additional	O
putative	O
therapeutic	O
and	O
preventive	O
targets	O
for	O
this	O
disease	O
.	O

Brick1	B-Gene_or_gene_product
is	O
an	O
essential	O
regulator	O
of	O
actin	B-Gene_or_gene_product
cytoskeleton	B-Cellular_component
required	O
for	O
embryonic	B-Developing_anatomical_structure
development	O
and	O
cell	B-Cell
transformation	O
.	O

Brick1	B-Gene_or_gene_product
(	O
Brk1	B-Gene_or_gene_product
)	O
is	O
the	O
less	O
-	O
studied	O
component	O
of	O
the	O
Wave	B-Gene_or_gene_product
/	O
Scar	B-Gene_or_gene_product
pathway	O
involved	O
in	O
the	O
branched	O
nucleation	O
of	O
actin	B-Gene_or_gene_product
fibers	B-Cellular_component
.	O

The	O
clinical	O
relevance	O
of	O
Brk1	B-Gene_or_gene_product
is	O
emphasized	O
by	O
correlative	O
data	O
showing	O
that	O
Von	O
Hippel	O
-	O
Lindau	O
(	O
VHL	O
)	O
patients	B-Organism
that	O
also	O
lose	O
the	O
BRK1	B-Gene_or_gene_product
gene	O
are	O
protected	O
against	O
the	O
development	O
of	O
tumors	B-Cancer
.	O

This	O
contrasts	O
with	O
recent	O
evidence	O
suggesting	O
that	O
the	O
Wave	B-Gene_or_gene_product
complex	O
may	O
function	O
as	O
an	O
invasion	O
suppressor	O
in	O
epithelial	B-Cancer
cancers	I-Cancer
.	O

Here	O
,	O
we	O
show	O
that	O
the	O
downregulation	O
of	O
Brk1	B-Gene_or_gene_product
results	O
in	O
abnormal	O
actin	B-Gene_or_gene_product
stress	B-Cellular_component
fiber	I-Cellular_component
formation	O
and	O
vinculin	B-Gene_or_gene_product
distribution	O
and	O
loss	O
of	O
Arp2	B-Gene_or_gene_product
/	I-Gene_or_gene_product
3	I-Gene_or_gene_product
and	O
Wave	B-Gene_or_gene_product
proteins	O
at	O
the	O
cellular	B-Cell
protrusions	O
.	O

Brk1	B-Gene_or_gene_product
is	O
required	O
for	O
cell	B-Cell
proliferation	O
and	O
cell	B-Cell
transformation	O
by	O
oncogenes	O
.	O

In	O
addition	O
,	O
Brk1	B-Gene_or_gene_product
downregulation	O
results	O
in	O
defective	O
directional	O
migration	O
and	O
invasive	O
growth	O
in	O
renal	B-Cell
cell	I-Cell
carcinoma	I-Cell
cells	I-Cell
as	O
well	O
as	O
in	O
other	O
tumor	B-Cell
cell	I-Cell
types	O
.	O

Finally	O
,	O
genetic	O
ablation	O
of	O
Brk1	B-Gene_or_gene_product
results	O
in	O
dramatic	O
defects	O
in	O
embryo	B-Developing_anatomical_structure
compaction	O
and	O
development	O
,	O
suggesting	O
an	O
essential	O
role	O
for	O
this	O
protein	O
in	O
actin	B-Gene_or_gene_product
dynamics	O
.	O

Thus	O
,	O
genetic	O
loss	O
or	O
inhibition	O
of	O
BRK1	B-Gene_or_gene_product
is	O
likely	O
to	O
be	O
protective	O
against	O
tumor	B-Cancer
development	O
due	O
to	O
proliferation	O
and	O
motility	O
defects	O
in	O
affected	O
cells	B-Cell
.	O

Ras	B-Gene_or_gene_product
-	I-Gene_or_gene_product
association	I-Gene_or_gene_product
domain	I-Gene_or_gene_product
family	I-Gene_or_gene_product
1C	I-Gene_or_gene_product
protein	O
promotes	O
breast	B-Cell
cancer	I-Cell
cell	I-Cell
migration	O
and	O
attenuates	O
apoptosis	O
.	O

BACKGROUND	O
:	O
The	O
Ras	B-Gene_or_gene_product
association	I-Gene_or_gene_product
domain	I-Gene_or_gene_product
family	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
RASSF1	B-Gene_or_gene_product
)	O
gene	O
is	O
a	O
Ras	B-Gene_or_gene_product
effector	O
encoding	O
two	O
major	O
mRNA	O
forms	O
,	O
RASSF1A	B-Gene_or_gene_product
and	O
RASSF1C	B-Gene_or_gene_product
,	O
derived	O
by	O
alternative	O
promoter	O
selection	O
and	O
alternative	O
mRNA	O
splicing	O
.	O

RASSF1A	B-Gene_or_gene_product
is	O
a	O
tumor	B-Cancer
suppressor	O
gene	O
.	O

However	O
,	O
very	O
little	O
is	O
known	O
about	O
the	O
function	O
of	O
RASSF1C	B-Gene_or_gene_product
both	O
in	O
normal	O
and	O
transformed	O
cells	B-Cell
.	O

METHODS	O
:	O
Gene	O
silencing	O
and	O
over	O
-	O
expression	O
techniques	O
were	O
used	O
to	O
modulate	O
RASSF1C	B-Gene_or_gene_product
expression	O
in	O
human	B-Organism
breast	B-Cell
cancer	I-Cell
cells	I-Cell
.	O

Affymetrix	O
-	O
microarray	O
analysis	O
was	O
performed	O
using	O
T47D	B-Cell
cells	I-Cell
over	O
-	O
expressing	O
RASSF1C	B-Gene_or_gene_product
to	O
identify	O
RASSF1C	B-Gene_or_gene_product
target	O
genes	O
.	O

RT	O
-	O
PCR	O
and	O
western	O
blot	O
techniques	O
were	O
used	O
to	O
validate	O
target	O
gene	O
expression	O
.	O

Cell	B-Cell
invasion	O
and	O
apoptosis	O
assays	O
were	O
also	O
performed	O
.	O

RESULTS	O
:	O
In	O
this	O
article	O
,	O
we	O
report	O
the	O
effects	O
of	O
altering	O
RASSF1C	B-Gene_or_gene_product
expression	O
in	O
human	B-Organism
breast	B-Cell
cancer	I-Cell
cells	I-Cell
.	O

We	O
found	O
that	O
silencing	O
RASSF1C	B-Gene_or_gene_product
mRNA	O
in	O
breast	B-Cell
cancer	I-Cell
cell	I-Cell
lines	I-Cell
(	O
MDA	B-Cell
-	I-Cell
MB231	I-Cell
and	O
T47D	B-Cell
)	O
caused	O
a	O
small	O
but	O
significant	O
decrease	O
in	O
cell	B-Cell
proliferation	O
.	O

Conversely	O
,	O
inducible	O
over	O
-	O
expression	O
of	O
RASSF1C	B-Gene_or_gene_product
in	O
breast	B-Cell
cancer	I-Cell
cells	I-Cell
(	O
MDA	B-Cell
-	I-Cell
MB231	I-Cell
and	O
T47D	B-Cell
)	O
resulted	O
in	O
a	O
small	O
increase	O
in	O
cell	B-Cell
proliferation	O
.	O

We	O
also	O
report	O
on	O
the	O
identification	O
of	O
novel	O
RASSF1C	B-Gene_or_gene_product
target	O
genes	O
.	O

RASSF1C	B-Gene_or_gene_product
down	O
-	O
regulates	O
several	O
pro	O
-	O
apoptotic	O
and	O
tumor	B-Cancer
suppressor	O
genes	O
and	O
up	O
-	O
regulates	O
several	O
growth	O
promoting	O
genes	O
in	O
breast	B-Cell
cancer	I-Cell
cells	I-Cell
.	O

We	O
further	O
show	O
that	O
down	O
-	O
regulation	O
of	O
caspase	B-Gene_or_gene_product
3	I-Gene_or_gene_product
via	O
overexpression	O
of	O
RASSF1C	B-Gene_or_gene_product
reduces	O
breast	B-Cell
cancer	I-Cell
cells	I-Cell
'	O
sensitivity	O
to	O
the	O
apoptosis	O
inducing	O
agent	O
,	O
etoposide	B-Simple_chemical
.	O

Furthermore	O
,	O
we	O
found	O
that	O
RASSF1C	B-Gene_or_gene_product
over	O
-	O
expression	O
enhances	O
T47D	B-Cell
cell	I-Cell
invasion	O
/	O
migration	O
in	O
vitro	O
.	O

CONCLUSION	O
:	O
Together	O
,	O
our	O
findings	O
suggest	O
that	O
RASSF1C	B-Gene_or_gene_product
,	O
unlike	O
RASSF1A	B-Gene_or_gene_product
,	O
is	O
not	O
a	O
tumor	B-Cancer
suppressor	O
,	O
but	O
instead	O
may	O
play	O
a	O
role	O
in	O
stimulating	O
metastasis	O
and	O
survival	O
in	O
breast	B-Cell
cancer	I-Cell
cells	I-Cell
.	O

Role	O
of	O
stromal	B-Cell
myofibroblasts	I-Cell
in	O
invasive	O
breast	B-Cancer
cancer	I-Cancer
:	O
stromal	B-Cell
expression	O
of	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
smooth	I-Gene_or_gene_product
muscle	I-Gene_or_gene_product
actin	I-Gene_or_gene_product
correlates	O
with	O
worse	O
clinical	O
outcome	O
.	O

BACKGROUND	O
:	O
Recently	O
,	O
the	O
desmoplastic	O
reaction	O
has	O
been	O
implicated	O
as	O
having	O
an	O
important	O
function	O
in	O
epithelial	B-Cancer
solid	I-Cancer
tumor	I-Cancer
biology	O
.	O

There	O
have	O
been	O
no	O
reports	O
showing	O
the	O
relativity	O
of	O
invasive	O
breast	B-Cancer
cancer	I-Cancer
and	O
the	O
desmoplastic	O
reaction	O
by	O
a	O
quantitative	O
analysis	O
of	O
the	O
myofibroblasts	B-Cell
that	O
were	O
an	O
important	O
player	O
in	O
the	O
desmoplastic	O
reaction	O
.	O

The	O
purpose	O
of	O
this	O
study	O
was	O
to	O
immunohistochemically	O
investigate	O
the	O
correlation	O
between	O
the	O
desmoplastic	O
reaction	O
and	O
the	O
clinicopathology	O
of	O
invasive	O
breast	B-Cancer
cancer	I-Cancer
.	O

METHODS	O
:	O
The	O
study	O
included	O
60	O
patients	B-Organism
with	O
a	O
known	O
prognosis	O
of	O
invasive	B-Cancer
breast	I-Cancer
cancer	I-Cancer
.	O

We	O
quantified	O
the	O
expression	O
of	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
SMA	I-Gene_or_gene_product
as	O
a	O
marker	O
of	O
myofibroblasts	B-Cell
in	O
the	O
invasive	O
breast	B-Cancer
cancer	I-Cancer
.	O

After	O
staining	O
samples	O
for	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
SMA	I-Gene_or_gene_product
,	O
their	O
expression	O
was	O
extracted	O
and	O
quantified	O
as	O
a	O
relative	O
percentage	O
by	O
computer	O
-	O
assisted	O
image	O
analysis	O
.	O

RESULTS	O
:	O
There	O
was	O
relatively	O
wide	O
variation	O
in	O
the	O
expression	O
of	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
SMA	I-Gene_or_gene_product
with	O
the	O
percentage	O
of	O
the	O
area	O
from	O
0	O
.	O
68	O
to	O
28	O
.	O
15	O
%	O
(	O
mean	O
8	O
.	O
48	O
+	O
/	O
-	O
5	O
.	O
40	O
%	O
)	O
.	O

The	O
metastasis	O
group	O
showed	O
significantly	O
higher	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
SMA	I-Gene_or_gene_product
expression	O
compared	O
with	O
the	O
no	O
metastasis	O
group	O
(	O
p	O
<	O
0	O
.	O
001	O
)	O
.	O

When	O
the	O
patients	B-Organism
were	O
divided	O
into	O
two	O
groups	O
according	O
to	O
their	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
SMA	I-Gene_or_gene_product
expression	O
using	O
a	O
cutoff	O
point	O
at	O
the	O
mean	O
value	O
of	O
8	O
.	O
48	O
%	O
,	O
the	O
high	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
SMA	I-Gene_or_gene_product
group	O
had	O
a	O
significantly	O
poorer	O
overall	O
survival	O
rate	O
(	O
p	O
<	O
0	O
.	O
001	O
)	O
.	O

Multivariate	O
analysis	O
demonstrated	O
that	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
SMA	I-Gene_or_gene_product
and	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
were	O
identified	O
as	O
independent	O
predictive	O
factors	O
of	O
metastasis	O
.	O

CONCLUSION	O
:	O
Myofibroblasts	B-Cell
represent	O
an	O
important	O
prognostic	O
factor	O
for	O
invasive	O
growth	O
that	O
is	O
translated	O
into	O
a	O
poor	O
clinical	O
prognosis	O
for	O
patients	B-Organism
with	O
invasive	B-Cancer
breast	I-Cancer
cancer	I-Cancer
.	O

The	O
disintegrin	B-Gene_or_gene_product
-	O
like	O
and	O
cysteine	B-Amino_acid
-	O
rich	O
domains	O
of	O
ADAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
mediate	O
interactions	O
between	O
melanoma	B-Cell
cells	I-Cell
and	O
fibroblasts	B-Cell
.	O

A	O
characteristic	O
of	O
malignant	B-Cell
cells	I-Cell
is	O
their	O
capacity	O
to	O
invade	O
their	O
surrounding	O
and	O
to	O
metastasize	O
to	O
distant	O
organs	B-Organ
.	O

During	O
these	O
processes	O
,	O
proteolytic	O
activities	O
of	O
tumor	B-Cell
and	O
stromal	B-Cell
cells	I-Cell
modify	O
the	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
to	O
produce	O
a	O
microenvironment	O
suitable	O
for	O
their	O
growth	O
and	O
migration	O
.	O

In	O
recent	O
years	O
the	O
family	O
of	O
ADAM	B-Gene_or_gene_product
proteases	O
has	O
been	O
ascribed	O
important	O
roles	O
in	O
these	O
processes	O
.	O

ADAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
is	O
expressed	O
in	O
human	B-Organism
melanoma	B-Cancer
at	O
the	O
tumor	B-Tissue
-	I-Tissue
stroma	I-Tissue
border	I-Tissue
where	O
direct	O
or	O
indirect	O
interactions	O
between	O
tumor	B-Cell
cells	I-Cell
and	O
fibroblasts	B-Cell
occur	O
.	O

To	O
analyze	O
the	O
role	O
of	O
ADAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
for	O
the	O
interaction	O
between	O
melanoma	B-Cell
cells	I-Cell
and	O
stromal	B-Cell
fibroblasts	I-Cell
,	O
we	O
produced	O
the	O
recombinant	O
disintegrin	B-Gene_or_gene_product
-	O
like	O
and	O
cysteine	B-Amino_acid
-	O
rich	O
domain	O
of	O
ADAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
(	O
DC	O
-	O
9	O
)	O
.	O

Melanoma	B-Cell
cells	I-Cell
and	O
human	B-Organism
fibroblasts	B-Cell
adhered	O
to	O
immobilized	O
DC	O
-	O
9	O
in	O
a	O
Mn	B-Simple_chemical
(	I-Simple_chemical
2	I-Simple_chemical
+	I-Simple_chemical
)	I-Simple_chemical
-	O
dependent	O
fashion	O
suggesting	O
an	O
integrin	B-Gene_or_gene_product
-	O
mediated	O
process	O
.	O

Inhibition	O
studies	O
showed	O
that	O
adhesion	O
of	O
fibroblasts	B-Cell
was	O
mediated	O
by	O
several	O
beta1	B-Gene_or_gene_product
integrin	I-Gene_or_gene_product
receptors	O
independent	O
of	O
the	O
RGD	O
and	O
ECD	O
recognition	O
motif	O
.	O

Furthermore	O
,	O
interaction	O
of	O
fibroblasts	B-Cell
and	O
high	B-Cell
invasive	I-Cell
melanoma	I-Cell
cells	I-Cell
with	O
soluble	O
recombinant	O
DC	O
-	O
9	O
resulted	O
in	O
enhanced	O
expression	O
of	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
.	O

Silencing	O
of	O
ADAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
in	O
melanoma	B-Cell
cells	I-Cell
significantly	O
reduced	O
cell	B-Cell
adhesion	O
to	O
fibroblasts	B-Cell
.	O

Ablation	O
of	O
ADAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
in	O
fibroblasts	B-Cell
almost	O
completely	O
abolished	O
these	O
cellular	B-Cell
interactions	O
and	O
melanoma	B-Cell
cell	I-Cell
invasion	O
in	O
vitro	O
.	O

In	O
summary	O
,	O
these	O
results	O
suggest	O
that	O
ADAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
expression	O
plays	O
an	O
important	O
role	O
in	O
mediating	O
cell	B-Cell
-	O
cell	B-Cell
contacts	O
between	O
fibroblasts	B-Cell
and	O
melanoma	B-Cell
cells	I-Cell
and	O
that	O
these	O
interactions	O
contribute	O
to	O
proteolytic	O
activities	O
required	O
during	O
invasion	O
of	O
melanoma	B-Cell
cells	I-Cell
.	O

Emerging	O
metabolic	O
targets	O
in	O
cancer	B-Cancer
therapy	O
.	O

Cancer	B-Cell
cells	I-Cell
are	O
different	O
from	O
normal	O
cells	B-Cell
in	O
their	O
metabolic	O
properties	O
.	O

Normal	O
cells	B-Cell
mostly	O
rely	O
on	O
mitochondrial	B-Cellular_component
oxidative	O
phosphorylation	O
to	O
produce	O
energy	O
.	O

In	O
contrast	O
,	O
cancer	B-Cell
cells	I-Cell
depend	O
mostly	O
on	O
glycolysis	O
,	O
the	O
aerobic	O
breakdown	O
of	O
glucose	B-Simple_chemical
into	O
ATP	B-Simple_chemical
.	O

This	O
altered	O
energy	O
dependency	O
is	O
known	O
as	O
the	O
"	O
Warburg	O
effect	O
"	O
and	O
is	O
a	O
hallmark	O
of	O
cancer	B-Cell
cells	I-Cell
.	O

In	O
recent	O
years	O
,	O
investigating	O
the	O
metabolic	O
changes	O
within	O
cancer	B-Cell
cells	I-Cell
has	O
been	O
a	O
rapidly	O
growing	O
area	B-Cancer
.	O

Emerging	O
evidence	O
shows	O
that	O
oncogenes	O
that	O
drive	O
the	O
cancer	B-Cancer
-	O
promoting	O
signals	O
also	O
drive	O
the	O
altered	O
metabolism	O
.	O

Although	O
the	O
exact	O
mechanisms	O
underlying	O
the	O
Warburg	O
effect	O
are	O
unclear	O
,	O
the	O
existing	O
evidence	O
suggests	O
that	O
increased	O
glycolysis	O
plays	O
an	O
important	O
role	O
in	O
support	O
malignant	O
behavior	O
of	O
cancer	B-Cell
cells	I-Cell
.	O

A	O
thorough	O
understanding	O
of	O
the	O
unique	O
metabolism	O
of	O
cancer	B-Cell
cells	I-Cell
will	O
help	O
to	O
design	O
of	O
more	O
effective	O
drugs	O
targeting	O
metabolic	O
pathways	O
,	O
which	O
will	O
greatly	O
impact	O
the	O
capacity	O
to	O
effectively	O
treat	O
cancer	B-Cancer
patients	B-Organism
.	O

Here	O
we	O
provide	O
an	O
overview	O
of	O
the	O
current	O
understanding	O
of	O
the	O
Warburg	O
effect	O
upon	O
tumor	B-Cell
cell	I-Cell
growth	O
and	O
survival	O
,	O
and	O
discussion	O
on	O
the	O
potential	O
metabolic	O
targets	O
for	O
cancer	B-Cancer
therapy	O
.	O

MTSS1	B-Gene_or_gene_product
:	O
a	O
multifunctional	O
protein	O
and	O
its	O
role	O
in	O
cancer	B-Cancer
invasion	O
and	O
metastasis	O
.	O

MTSS1	B-Gene_or_gene_product
(	O
metastasis	B-Gene_or_gene_product
suppressor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
was	O
first	O
identified	O
as	O
a	O
metastasis	O
suppressor	O
missing	O
in	O
metastatic	O
bladder	B-Cell
carcinoma	I-Cell
cell	I-Cell
lines	I-Cell
.	O

The	O
down	O
-	O
regulation	O
of	O
MTSS1	B-Gene_or_gene_product
that	O
may	O
be	O
caused	O
by	O
DNA	B-Cellular_component
methylation	O
was	O
also	O
observed	O
in	O
many	O
other	O
types	O
of	O
cancer	B-Cancer
.	O

While	O
accumlating	O
evidence	O
for	O
the	O
function	O
of	O
MTSS1	B-Gene_or_gene_product
support	O
the	O
concept	O
that	O
it	O
is	O
unlikely	O
to	O
be	O
a	O
metastasis	O
suppressor	O
,	O
but	O
actually	O
acts	O
as	O
a	O
scaffold	O
protein	O
that	O
interacts	O
with	O
multiple	O
partners	O
to	O
regulate	O
actin	B-Gene_or_gene_product
dynamics	O
.	O

It	O
has	O
also	O
been	O
demonstrated	O
that	O
MTSS1	B-Gene_or_gene_product
is	O
involved	O
in	O
the	O
Shh	B-Gene_or_gene_product
signaling	O
pathway	O
in	O
the	O
developing	O
hair	B-Multi-tissue_structure
follicle	I-Multi-tissue_structure
and	O
in	O
basal	B-Cancer
cell	I-Cancer
carcinomas	I-Cancer
of	O
the	O
skin	B-Organ
.	O

Such	O
evidence	O
indicates	O
that	O
MTSS1	B-Gene_or_gene_product
as	O
a	O
multiple	O
functional	O
molecular	O
player	O
and	O
has	O
an	O
important	O
role	O
in	O
development	O
,	O
carcinogenesis	O
and	O
metastasis	O
.	O

However	O
,	O
the	O
biochemical	O
mechanisms	O
by	O
which	O
MTSS1	B-Gene_or_gene_product
functions	O
in	O
cells	B-Cell
and	O
the	O
physiological	O
role	O
of	O
this	O
protein	O
in	O
animals	O
remain	O
largely	O
unknown	O
.	O

In	O
this	O
review	O
,	O
we	O
will	O
discuss	O
the	O
current	O
knowledge	O
of	O
MTSS1	B-Gene_or_gene_product
'	O
s	O
role	O
in	O
cancer	B-Cancer
metastasis	O
,	O
carcinogenesis	O
,	O
and	O
development	O
.	O

The	O
clinical	O
significance	O
of	O
MTSS1	B-Gene_or_gene_product
will	O
also	O
be	O
discussed	O
.	O

Ras	B-Gene_or_gene_product
p21	I-Gene_or_gene_product
onco	O
-	O
protein	O
in	O
the	O
sera	B-Organism_substance
of	O
mice	B-Organism
carrying	O
an	O
experimentally	O
induced	O
tumor	B-Cancer
and	O
in	O
human	B-Organism
cancer	B-Cancer
patients	B-Organism
.	O

We	O
have	O
established	O
a	O
model	O
system	O
to	O
detect	O
the	O
presence	O
of	O
ras	B-Gene_or_gene_product
p21	I-Gene_or_gene_product
in	O
the	O
sera	B-Organism_substance
of	O
Balb	B-Organism
/	I-Organism
c	I-Organism
mice	I-Organism
carrying	O
tumors	B-Cancer
induced	O
by	O
a	O
mouse	B-Organism
cell	B-Cell
line	I-Cell
transformed	O
with	O
the	O
Harvey	B-Organism
murine	I-Organism
sarcoma	I-Organism
virus	I-Organism
in	O
the	O
presence	O
of	O
a	O
helper	O
Friend	B-Organism
murine	I-Organism
leukemia	I-Organism
virus	I-Organism
.	O

As	O
determined	O
by	O
ELISA	O
and	O
immunoblot	O
assays	O
,	O
ras	B-Gene_or_gene_product
p21	I-Gene_or_gene_product
in	O
the	O
serum	B-Organism_substance
increased	O
with	O
increased	O
tumor	B-Cancer
growth	O
.	O

Since	O
ras	B-Gene_or_gene_product
genes	O
have	O
been	O
found	O
to	O
be	O
frequently	O
activated	O
in	O
human	B-Organism
tumours	B-Cancer
,	O
we	O
examined	O
the	O
levels	O
of	O
ras	B-Gene_or_gene_product
p21	I-Gene_or_gene_product
in	O
the	O
sera	B-Organism_substance
of	O
a	O
variety	O
of	O
human	B-Organism
cancer	B-Cancer
patients	B-Organism
.	O

In	O
only	O
3	O
out	O
of	O
13	O
cases	O
,	O
representing	O
patients	B-Organism
with	O
adenocarcinomas	B-Cancer
of	O
the	O
stomach	B-Organ
receiving	O
chemotherapy	O
,	O
was	O
ras	B-Gene_or_gene_product
p21	I-Gene_or_gene_product
detected	O
at	O
elevated	O
levels	O
,	O
whereas	O
in	O
patients	B-Organism
with	O
the	O
following	O
types	O
of	O
cancer	B-Cancer
no	O
substantial	O
change	O
in	O
serum	B-Organism_substance
ras	B-Gene_or_gene_product
p21	I-Gene_or_gene_product
was	O
observed	O
;	O
nine	O
with	O
breast	B-Cancer
,	O
5	O
colon	B-Cancer
,	O
5	O
lung	B-Cancer
,	O
5	O
ovarian	B-Cancer
and	O
5	O
hepatocellular	B-Cancer
carcinomas	I-Cancer
.	O

Serum	B-Organism_substance
soluble	O
vascular	B-Gene_or_gene_product
adhesion	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
is	O
a	O
valuable	O
prognostic	O
marker	O
in	O
gastric	B-Cancer
cancer	I-Cancer
.	O

BACKGROUND	O
:	O
Vascular	B-Gene_or_gene_product
adhesion	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
VAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
regulates	O
leukocyte	B-Cell
tissue	I-Cell
infiltration	O
.	O

Elevated	O
serum	B-Organism_substance
soluble	O
VAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
sVAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
levels	O
occur	O
in	O
certain	O
diseases	O
having	O
an	O
inflammatory	O
component	O
.	O

We	O
previously	O
showed	O
in	O
colorectal	B-Cancer
cancer	I-Cancer
that	O
sVAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
expression	O
is	O
significantly	O
higher	O
relative	O
to	O
controls	O
,	O
and	O
this	O
decreased	O
expression	O
is	O
associated	O
with	O
poor	O
prognosis	O
and	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
and	O
liver	B-Organ
metastasis	O
.	O

However	O
,	O
sVAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
expression	O
has	O
not	O
been	O
described	O
for	O
gastric	B-Cancer
cancer	I-Cancer
.	O

This	O
study	O
determines	O
the	O
relationship	O
between	O
preoperative	O
serum	B-Organism_substance
sVAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
levels	O
and	O
clinicopathological	O
features	O
and	O
prognosis	O
in	O
gastric	B-Cancer
cancer	I-Cancer
.	O

METHODS	O
:	O
Preoperative	O
serum	B-Organism_substance
was	O
collected	O
from	O
107	O
gastric	B-Cancer
cancer	I-Cancer
patients	B-Organism
and	O
33	O
normal	O
controls	O
.	O

sVAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
levels	O
were	O
assayed	O
by	O
enzyme	O
-	O
linked	O
immunosorbent	O
assay	O
.	O

RESULTS	O
:	O
The	O
mean	O
sVAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
level	O
for	O
cancer	B-Cancer
patients	B-Organism
was	O
significantly	O
higher	O
relative	O
to	O
controls	O
,	O
and	O
decreased	O
with	O
disease	B-Cancer
progression	O
.	O

Tumor	B-Cancer
size	O
,	O
serosal	B-Multi-tissue_structure
invasion	O
,	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
,	O
peritoneal	B-Multi-tissue_structure
dissemination	O
,	O
and	O
TNM	O
classification	O
was	O
significantly	O
correlated	O
with	O
sVAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
level	O
.	O

sVAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
is	O
also	O
an	O
independent	O
predictive	O
marker	O
for	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
.	O

Patients	B-Organism
having	O
low	O
sVAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
levels	O
had	O
significantly	O
poorer	O
prognosis	O
relative	O
to	O
patients	B-Organism
having	O
elevated	O
sVAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
in	O
all	O
or	O
stages	O
I	O
-	O
III	O
gastric	B-Cancer
cancer	I-Cancer
patients	B-Organism
,	O
respectively	O
.	O

CONCLUSIONS	O
:	O
Low	O
sVAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
levels	O
are	O
associated	O
with	O
poor	O
prognosis	O
in	O
gastric	B-Cancer
cancer	I-Cancer
.	O

Determining	O
sVAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
levels	O
may	O
be	O
valuable	O
for	O
predicting	O
prognosis	O
and	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
.	O

Metabolic	O
regulation	O
by	O
p53	B-Gene_or_gene_product
.	O

We	O
are	O
increasingly	O
aware	O
that	O
cellular	B-Cell
metabolism	O
plays	O
a	O
vital	O
role	O
in	O
diseases	O
such	O
as	O
cancer	B-Cancer
,	O
and	O
that	O
p53	B-Gene_or_gene_product
is	O
an	O
important	O
regulator	O
of	O
metabolic	O
pathways	O
.	O

By	O
transcriptional	O
activation	O
and	O
other	O
means	O
,	O
p53	B-Gene_or_gene_product
is	O
able	O
to	O
contribute	O
to	O
the	O
regulation	O
of	O
glycolysis	O
,	O
oxidative	O
phosphorylation	O
,	O
glutaminolysis	O
,	O
insulin	B-Gene_or_gene_product
sensitivity	O
,	O
nucleotide	B-Simple_chemical
biosynthesis	O
,	O
mitochondrial	B-Cellular_component
integrity	O
,	O
fatty	B-Simple_chemical
acid	I-Simple_chemical
oxidation	O
,	O
antioxidant	B-Simple_chemical
response	O
,	O
autophagy	O
and	O
mTOR	B-Gene_or_gene_product
signalling	O
.	O

The	O
ability	O
to	O
positively	O
and	O
negatively	O
regulate	O
many	O
of	O
these	O
pathways	O
,	O
combined	O
with	O
feedback	O
signalling	O
from	O
these	O
pathways	O
to	O
p53	B-Gene_or_gene_product
,	O
demonstrates	O
the	O
reciprocal	O
and	O
flexible	O
nature	O
of	O
the	O
regulation	O
,	O
facilitating	O
a	O
diverse	O
range	O
of	O
responses	O
to	O
metabolic	O
stress	O
.	O

Intriguingly	O
,	O
metabolic	O
stress	O
triggers	O
primarily	O
an	O
adaptive	O
(	O
rather	O
than	O
pro	O
-	O
apoptotic	O
)	O
p53	B-Gene_or_gene_product
response	O
,	O
and	O
p53	B-Gene_or_gene_product
is	O
emerging	O
as	O
an	O
important	O
regulator	O
of	O
metabolic	O
homeostasis	O
.	O

A	O
better	O
understanding	O
of	O
how	O
p53	B-Gene_or_gene_product
coordinates	O
metabolic	O
adaptation	O
will	O
facilitate	O
the	O
identification	O
of	O
novel	O
therapeutic	O
targets	O
and	O
will	O
also	O
illuminate	O
the	O
wider	O
role	O
of	O
p53	B-Gene_or_gene_product
in	O
human	B-Organism
biology	O
.	O

Use	O
of	O
DNA	B-Cellular_component
microarray	O
and	O
small	O
animal	O
positron	O
emission	O
tomography	O
in	O
preclinical	O
drug	O
evaluation	O
of	O
RAF265	B-Simple_chemical
,	O
a	O
novel	O
B	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Raf	I-Gene_or_gene_product
/	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
inhibitor	O
.	O

Positron	O
emission	O
tomography	O
(	O
PET	O
)	O
imaging	O
has	O
become	O
a	O
useful	O
tool	O
for	O
assessing	O
early	O
biologic	O
response	O
to	O
cancer	B-Cancer
therapy	O
and	O
may	O
be	O
particularly	O
useful	O
in	O
the	O
development	O
of	O
new	O
cancer	B-Cancer
therapeutics	O
.	O

RAF265	B-Simple_chemical
,	O
a	O
novel	O
B	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Raf	I-Gene_or_gene_product
/	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
inhibitor	O
,	O
was	O
evaluated	O
in	O
the	O
preclinical	O
setting	O
for	O
its	O
ability	O
to	O
inhibit	O
the	O
uptake	O
of	O
PET	O
tracers	O
in	O
the	O
A375M	O
(	O
B	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Raf	I-Gene_or_gene_product
(	O
V600E	O
)	O
)	O
human	O
melanoma	O
cell	O
line	O
.	O

RAF265	B-Simple_chemical
inhibited	O
2	B-Simple_chemical
-	I-Simple_chemical
deoxy	I-Simple_chemical
-	I-Simple_chemical
2	I-Simple_chemical
-	I-Simple_chemical
[	I-Simple_chemical
(	I-Simple_chemical
18	I-Simple_chemical
)	I-Simple_chemical
F	I-Simple_chemical
]	I-Simple_chemical
fluoro	I-Simple_chemical
-	I-Simple_chemical
d	I-Simple_chemical
-	I-Simple_chemical
glucose	I-Simple_chemical
(	O
FDG	B-Simple_chemical
)	O
accumulation	O
in	O
cell	B-Cell
culture	O
at	O
28	O
hours	O
in	O
a	O
dose	O
-	O
dependent	O
manner	O
.	O

RAF265	B-Simple_chemical
also	O
inhibited	O
FDG	B-Simple_chemical
accumulation	O
in	O
tumor	B-Cancer
xenografts	I-Cancer
after	O
1	O
day	O
of	O
drug	O
treatment	O
.	O

This	O
decrease	O
persisted	O
for	O
the	O
remaining	O
2	O
weeks	O
of	O
treatment	O
.	O

DNA	B-Cellular_component
microarray	O
analysis	O
of	O
treated	O
tumor	B-Cancer
xenografts	I-Cancer
revealed	O
significantly	O
decreased	O
expression	O
of	O
genes	O
regulating	O
glucose	B-Simple_chemical
and	O
thymidine	B-Simple_chemical
metabolism	O
and	O
revealed	O
changes	O
in	O
apoptotic	O
genes	O
,	O
suggesting	O
that	O
the	O
imaging	O
tracers	O
FDG	B-Simple_chemical
,	O
3	B-Simple_chemical
-	I-Simple_chemical
deoxy	I-Simple_chemical
-	I-Simple_chemical
3	I-Simple_chemical
-	I-Simple_chemical
[	I-Simple_chemical
(	I-Simple_chemical
18	I-Simple_chemical
)	I-Simple_chemical
F	I-Simple_chemical
]	I-Simple_chemical
fluorothymidine	I-Simple_chemical
,	O
and	O
annexin	B-Gene_or_gene_product
V	I-Gene_or_gene_product
could	O
serve	O
as	O
potential	O
imaging	O
biomarkers	O
for	O
RAF265	B-Simple_chemical
therapy	O
monitoring	O
.	O

We	O
concluded	O
that	O
RAF265	B-Simple_chemical
is	O
highly	O
efficacious	O
in	O
this	O
xenograft	B-Cancer
model	O
of	O
human	B-Organism
melanoma	B-Cancer
and	O
decreases	O
glucose	B-Simple_chemical
metabolism	O
as	O
measured	O
by	O
DNA	B-Cellular_component
microarray	O
analysis	O
,	O
cell	B-Cell
culture	O
assays	O
,	O
and	O
small	O
animal	O
FDG	B-Simple_chemical
PET	O
scans	O
as	O
early	O
as	O
1	O
day	O
after	O
treatment	O
.	O

Our	O
results	O
support	O
the	O
use	O
of	O
FDG	B-Simple_chemical
PET	O
in	O
clinical	O
trials	O
with	O
RAF265	B-Simple_chemical
to	O
assess	O
early	O
tumor	B-Cancer
response	O
.	O

DNA	B-Cellular_component
microarray	O
analysis	O
and	O
small	O
animal	O
PET	O
studies	O
may	O
be	O
used	O
as	O
complementary	O
technologies	O
in	O
drug	O
development	O
.	O

DNA	B-Cellular_component
microarray	O
analysis	O
allows	O
for	O
analysis	O
of	O
drug	O
effects	O
on	O
multiple	O
pathways	O
linked	O
to	O
cancer	B-Cancer
and	O
can	O
suggest	O
corresponding	O
imaging	O
tracers	O
for	O
further	O
analysis	O
as	O
biomarkers	O
of	O
tumor	B-Cancer
response	O
.	O

Systematic	O
proteomic	O
analysis	O
of	O
human	B-Organism
hepotacellular	B-Cell
carcinoma	I-Cell
cells	I-Cell
reveals	O
molecular	O
pathways	O
and	O
networks	O
involved	O
in	O
metastasis	O
.	O

Systematic	O
proteomic	O
studying	O
of	O
the	O
mechanism	O
of	O
hepatocellular	B-Cancer
carcinoma	I-Cancer
(	O
HCC	B-Cancer
)	O
metastasis	O
remains	O
challenging	O
.	O

We	O
performed	O
comparative	O
proteomic	O
and	O
pathway	O
analysis	O
of	O
four	O
human	B-Organism
metastatic	B-Cell
HCC	I-Cell
cell	I-Cell
lines	I-Cell
to	O
identify	O
metastasis	O
-	O
associated	O
proteins	O
.	O

These	O
HCC	B-Cell
cell	I-Cell
lines	I-Cell
had	O
a	O
similar	O
genetic	O
background	O
but	O
with	O
an	O
increasing	O
potential	O
of	O
metastasis	O
.	O

Using	O
a	O
combination	O
of	O
two	O
dimensional	O
electrophoresis	O
(	O
2	O
-	O
DE	O
)	O
and	O
MALDI	O
-	O
TOF	O
mass	O
spectrometry	O
,	O
a	O
total	O
of	O
125	O
proteins	O
and	O
their	O
post	O
-	O
translational	O
modification	O
forms	O
or	O
isoforms	O
were	O
found	O
to	O
be	O
differentially	O
expressed	O
in	O
the	O
cell	B-Cell
lines	I-Cell
.	O

Among	O
them	O
,	O
29	O
were	O
gradually	O
up	O
-	O
regulated	O
whereas	O
17	O
were	O
down	O
-	O
regulated	O
with	O
increasing	O
metastatic	O
potential	O
.	O

Instead	O
of	O
a	O
traditional	O
single	O
-	O
gene	O
readout	O
,	O
global	O
bioinformatics	O
analysis	O
was	O
carried	O
out	O
,	O
which	O
revealed	O
that	O
the	O
glycolysis	O
pathway	O
was	O
the	O
most	O
significantly	O
enriched	O
pathway	O
.	O

The	O
heat	B-Gene_or_gene_product
shock	I-Gene_or_gene_product
proteins	I-Gene_or_gene_product
(	O
HSPs	B-Gene_or_gene_product
)	O
centered	O
and	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
centered	O
networks	O
were	O
also	O
enriched	O
in	O
the	O
result	O
,	O
which	O
may	O
imply	O
the	O
key	O
function	O
of	O
inflaming	O
on	O
metastasis	O
.	O

Meanwhile	O
,	O
knockdown	O
of	O
HDGF	B-Gene_or_gene_product
,	O
an	O
up	O
-	O
regulated	O
protein	O
and	O
a	O
target	O
of	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
,	O
induced	O
cell	B-Cell
apoptosis	O
in	O
the	O
metastatic	O
HCC	B-Cell
cells	I-Cell
.	O

This	O
work	O
provides	O
a	O
demonstration	O
that	O
a	O
combination	O
of	O
bioinformatics	O
and	O
comparative	O
proteomics	O
can	O
help	O
in	O
finding	O
out	O
potential	O
biomarkers	O
associated	O
with	O
HCC	B-Cancer
metastasis	O
on	O
the	O
level	O
of	O
pathways	O
.	O

Induction	O
of	O
Id	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
by	O
FGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
involves	O
activity	O
of	O
EGR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
sensitizes	O
neuroblastoma	B-Cell
cells	I-Cell
to	O
cell	B-Cell
death	O
.	O

Inhibitor	B-Gene_or_gene_product
of	I-Gene_or_gene_product
differentiation	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
Id	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
is	O
a	O
member	O
of	O
helix	O
-	O
loop	O
-	O
helix	O
(	O
HLH	O
)	O
family	O
of	O
proteins	O
that	O
regulate	O
gene	O
transcription	O
through	O
their	O
inhibitory	O
binding	O
to	O
basic	O
-	O
HLH	O
transcription	O
factors	O
.	O

Similarly	O
to	O
other	O
members	O
of	O
this	O
family	O
,	O
Id	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
is	O
involved	O
in	O
the	O
repression	O
of	O
cell	B-Cell
differentiation	O
and	O
activation	O
of	O
cell	B-Cell
growth	O
.	O

The	O
dual	O
function	O
of	O
Id	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
inhibition	O
of	O
differentiation	O
,	O
and	O
stimulation	O
of	O
cell	B-Cell
proliferation	O
,	O
might	O
be	O
interdependent	O
,	O
as	O
cell	B-Cell
differentiation	O
is	O
generally	O
coupled	O
with	O
the	O
exit	O
from	O
the	O
cell	B-Cell
cycle	O
.	O

Fibroblast	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
(	O
FGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
)	O
has	O
been	O
reported	O
to	O
play	O
multiple	O
roles	O
in	O
different	O
biological	O
processes	O
during	O
development	O
of	O
the	O
central	B-Anatomical_system
nervous	I-Anatomical_system
system	I-Anatomical_system
(	O
CNS	B-Anatomical_system
)	O
.	O

In	O
addition	O
,	O
FGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
has	O
been	O
described	O
to	O
induce	O
"	O
neuronal	B-Cell
-	O
like	O
"	O
differentiation	O
and	O
trigger	O
apoptosis	O
in	O
neuroblastoma	B-Cell
SK	I-Cell
-	I-Cell
N	I-Cell
-	I-Cell
MC	I-Cell
cells	I-Cell
.	O

Although	O
regulation	O
of	O
Id	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
protein	O
by	O
several	O
mitogenic	O
factors	O
is	O
well	O
-	O
established	O
,	O
little	O
is	O
known	O
about	O
the	O
role	O
of	O
FGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
in	O
the	O
regulation	O
of	O
Id	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

Using	O
human	B-Organism
neuroblastoma	B-Cell
cell	I-Cell
line	I-Cell
,	O
SK	B-Cell
-	I-Cell
N	I-Cell
-	I-Cell
MC	I-Cell
,	O
we	O
found	O
that	O
treatment	O
of	O
these	O
cells	B-Cell
with	O
FGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
resulted	O
in	O
early	O
induction	O
of	O
both	O
Id	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
mRNA	O
and	O
protein	O
.	O

The	O
induction	O
occurs	O
within	O
1	O
h	O
from	O
FGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
treatment	O
and	O
is	O
mediated	O
by	O
ERK1	B-Gene_or_gene_product
/	I-Gene_or_gene_product
2	I-Gene_or_gene_product
pathway	O
,	O
which	O
in	O
turn	O
stimulates	O
expression	O
of	O
the	O
early	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
response	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
Egr	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
transcription	O
factor	O
.	O

We	O
also	O
demonstrate	O
direct	O
interaction	O
of	O
Egr	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
with	O
Id	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
promoter	O
in	O
vitro	O
and	O
in	O
cell	B-Cell
culture	I-Cell
.	O

Finally	O
,	O
inhibition	O
of	O
Id	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
expression	O
results	O
in	O
G	O
(	O
2	O
)	O
/	O
M	O
accumulation	O
of	O
FGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
-	O
treated	O
cells	B-Cell
and	O
delayed	O
cell	B-Cell
death	O
.	O

Vimentin	B-Gene_or_gene_product
is	O
preferentially	O
expressed	O
in	O
human	B-Organism
breast	B-Cancer
carcinomas	I-Cancer
with	O
low	O
estrogen	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
and	O
high	O
Ki	B-Gene_or_gene_product
-	I-Gene_or_gene_product
67	I-Gene_or_gene_product
growth	O
fraction	O
.	O

Vimentin	B-Gene_or_gene_product
expression	O
,	O
growth	O
fractions	O
(	O
GF	O
)	O
,	O
and	O
estrogen	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
(	O
ER	B-Gene_or_gene_product
)	O
levels	O
were	O
determined	O
for	O
90	O
untreated	O
primary	O
breast	B-Cancer
carcinomas	I-Cancer
.	O

Coexpression	O
of	O
keratin	B-Gene_or_gene_product
and	O
vimentin	B-Gene_or_gene_product
was	O
found	O
in	O
approximately	O
20	O
%	O
of	O
the	O
tumors	B-Cancer
regardless	O
of	O
menopausal	O
status	O
.	O

Vimentin	B-Gene_or_gene_product
was	O
expressed	O
preferentially	O
in	O
tumor	B-Cell
cells	I-Cell
of	O
high	B-Cancer
-	I-Cancer
grade	I-Cancer
ductal	I-Cancer
breast	I-Cancer
carcinomas	I-Cancer
(	O
15	O
of	O
28	O
histologic	O
grade	O
3	O
vs	O
.	O
0	O
of	O
40	O
grades	O
1	O
and	O
2	O
)	O
.	O

Vimentin	B-Gene_or_gene_product
expression	O
was	O
found	O
preferentially	O
in	O
tumors	B-Cancer
with	O
high	O
GF	O
(	O
greater	O
than	O
15	O
%	O
Ki	B-Gene_or_gene_product
-	I-Gene_or_gene_product
67	I-Gene_or_gene_product
positive	O
by	O
immunoperoxidase	B-Gene_or_gene_product
staining	O
)	O
and	O
low	O
ER	B-Gene_or_gene_product
levels	O
(	O
less	O
than	O
60	O
fmols	O
/	O
mg	O
protein	O
by	O
a	O
monoclonal	O
enzyme	O
immunoassay	O
)	O
.	O

Sixty	O
-	O
eight	O
percent	O
of	O
tumors	B-Cancer
in	O
this	O
group	O
were	O
vimentin	B-Gene_or_gene_product
positive	O
and	O
88	O
%	O
of	O
all	O
vimentin	B-Gene_or_gene_product
-	O
positive	O
tumors	B-Cancer
fell	O
into	O
this	O
category	O
.	O

More	O
than	O
50	O
%	O
of	O
the	O
tumor	B-Cell
cells	I-Cell
coexpressed	O
vimentin	B-Gene_or_gene_product
and	O
keratin	B-Gene_or_gene_product
.	O

Thus	O
,	O
vimentin	B-Gene_or_gene_product
expression	O
may	O
be	O
helpful	O
in	O
identifying	O
a	O
substantial	O
subset	O
of	O
ER	B-Gene_or_gene_product
-	O
independent	O
breast	B-Cancer
carcinomas	I-Cancer
with	O
poor	O
prognostic	O
indicators	O
.	O

[	O
Expression	O
of	O
Merlin	B-Gene_or_gene_product
protein	O
in	O
non	B-Cancer
-	I-Cancer
small	I-Cancer
cell	I-Cancer
lung	I-Cancer
carcinoma	I-Cancer
and	O
the	O
clinical	O
significance	O
]	O
.	O

OBJECTIVE	O
:	O
To	O
determine	O
the	O
expression	O
and	O
clinical	O
significance	O
of	O
Merlin	B-Gene_or_gene_product
protein	O
in	O
non	B-Cancer
-	I-Cancer
small	I-Cancer
cell	I-Cancer
lung	I-Cancer
cancer	I-Cancer
(	O
NSCLC	B-Cancer
)	O
.	O

METHODS	O
:	O
The	O
expression	O
of	O
Merlin	B-Gene_or_gene_product
protein	O
in	O
45	O
cases	O
of	O
NSCLC	B-Cancer
and	O
adjacent	O
tissue	B-Tissue
of	O
NSCLC	B-Cancer
and	O
normal	O
lung	B-Tissue
tissue	I-Tissue
was	O
checked	O
by	O
immunohistochemistry	O
.	O

The	O
relation	O
between	O
the	O
expression	O
of	O
Merlin	B-Gene_or_gene_product
protein	O
and	O
the	O
multiple	O
factors	O
of	O
pathological	O
type	O
,	O
gender	O
,	O
P	O
-	O
TNM	O
stage	O
,	O
differentiation	O
and	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
was	O
analyzed	O
.	O

RESULTS	O
:	O
The	O
expression	O
rates	O
of	O
Merlin	B-Gene_or_gene_product
protein	O
in	O
NSCLC	B-Cancer
and	O
normal	O
lung	B-Multi-tissue_structure
tissue	I-Multi-tissue_structure
sections	I-Multi-tissue_structure
were	O
73	O
.	O
33	O
%	O
and	O
15	O
.	O
56	O
%	O
,	O
respectively	O
(	O
P	O
<	O
0	O
.	O
05	O
)	O
.	O

The	O
expression	O
of	O
Merlin	B-Gene_or_gene_product
protein	O
was	O
not	O
associated	O
with	O
the	O
pathological	O
type	O
,	O
gender	O
,	O
P	O
-	O
TNM	O
stage	O
,	O
differentiation	O
and	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
(	O
P	O
>	O
0	O
.	O
05	O
)	O
.	O

CONCLUSION	O
:	O
Merlin	B-Gene_or_gene_product
protein	O
might	O
contribute	O
to	O
the	O
initiation	O
of	O
metastasis	O
of	O
NSCLC	B-Cancer
.	O

The	O
members	O
of	O
an	O
Epstein	B-Organism
-	I-Organism
Barr	I-Organism
virus	I-Organism
microRNA	O
cluster	O
cooperate	O
to	O
transform	O
B	B-Cell
lymphocytes	I-Cell
.	O

Epstein	B-Organism
-	I-Organism
Barr	I-Organism
virus	I-Organism
(	O
EBV	B-Organism
)	O
transforms	O
B	B-Cell
lymphocytes	I-Cell
through	O
the	O
expression	O
of	O
the	O
latent	O
viral	O
proteins	O
EBNA	B-Gene_or_gene_product
and	O
latent	B-Gene_or_gene_product
membrane	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
(	O
LMP	B-Gene_or_gene_product
)	O
.	O

Recently	O
,	O
it	O
has	O
become	O
apparent	O
that	O
microRNAs	O
(	O
miRNAs	O
)	O
also	O
contribute	O
to	O
EBV	B-Organism
'	O
s	O
oncogenic	O
properties	O
;	O
recombinant	B-Organism
EBVs	I-Organism
that	O
lack	O
the	O
BHRF1	B-Gene_or_gene_product
miRNA	O
cluster	O
display	O
a	O
reduced	O
ability	O
to	O
transform	O
B	B-Cell
lymphocytes	I-Cell
in	O
vitro	O
.	O

Furthermore	O
,	O
infected	O
cells	B-Cell
evince	O
a	O
marked	O
upregulation	O
of	O
the	O
EBNA	B-Gene_or_gene_product
genes	O
.	O

Using	O
recombinant	O
viruses	O
that	O
lack	O
only	O
one	O
member	O
of	O
the	O
cluster	O
,	O
we	O
now	O
show	O
that	O
all	O
three	O
BHRF1	B-Gene_or_gene_product
miRNAs	O
contribute	O
to	O
B	B-Cell
-	I-Cell
cell	I-Cell
transformation	O
.	O

Recombinants	O
that	O
lacked	O
miR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
BHRF1	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
or	O
miR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
BHRF1	I-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
displayed	O
enhanced	O
EBNA	B-Gene_or_gene_product
expression	O
initiated	O
at	O
the	O
Cp	O
and	O
Wp	O
promoters	O
.	O

Interestingly	O
,	O
we	O
find	O
that	O
the	O
deletion	O
of	O
miR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
BHRF1	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
reduced	O
the	O
expression	O
level	O
of	O
miR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
BHRF1	I-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
and	O
possibly	O
that	O
of	O
miR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
BHRF1	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
demonstrating	O
that	O
the	O
expression	O
of	O
one	O
miRNA	O
can	O
potentiate	O
the	O
expression	O
of	O
other	O
miRNAs	O
located	O
in	O
the	O
same	O
cluster	O
.	O

Therefore	O
,	O
the	O
phenotypic	O
traits	O
of	O
the	O
miR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
BHRF1	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
null	O
mutant	O
could	O
result	O
partly	O
from	O
reduced	O
miR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
BHRF1	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
miR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
BHRF1	I-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
expression	O
levels	O
.	O

Nevertheless	O
,	O
using	O
an	O
miR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
BHRF1	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
miR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
BHRF1	I-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
double	O
mutant	O
,	O
we	O
could	O
directly	O
assess	O
and	O
confirm	O
the	O
contribution	O
of	O
miR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
BHRF1	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
to	O
B	B-Cell
-	I-Cell
cell	I-Cell
transformation	O
.	O

Furthermore	O
,	O
we	O
found	O
that	O
the	O
potentiating	O
effect	O
of	O
miR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
BHRF1	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
on	O
miR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
BHRF1	I-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
synthesis	O
can	O
be	O
reproduced	O
with	O
simple	O
expression	O
plasmids	O
,	O
provided	O
that	O
both	O
miRNAs	O
are	O
processed	O
from	O
the	O
same	O
transcript	O
.	O

Therefore	O
,	O
this	O
enhancing	O
effect	O
does	O
not	O
result	O
from	O
an	O
idiosyncrasy	O
of	O
the	O
EBV	B-Organism
genome	O
but	O
rather	O
reflects	O
a	O
general	O
property	O
of	O
these	O
miRNAs	O
.	O

This	O
study	O
highlights	O
the	O
advantages	O
of	O
arranging	O
the	O
BHRF1	B-Gene_or_gene_product
miRNAs	O
in	O
clusters	O
:	O
it	O
allows	O
the	O
synchronous	O
and	O
synergistic	O
expression	O
of	O
genetic	O
elements	O
that	O
cooperate	O
to	O
transform	O
their	O
target	O
cells	B-Cell
.	O

Sunitinib	B-Simple_chemical
induces	O
apoptosis	O
in	O
pheochromocytoma	B-Cell
tumor	I-Cell
cells	I-Cell
by	O
inhibiting	O
VEGFR2	B-Gene_or_gene_product
/	O
Akt	B-Gene_or_gene_product
/	O
mTOR	B-Gene_or_gene_product
/	O
S6K1	B-Gene_or_gene_product
pathways	O
through	O
modulation	O
of	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
BAD	B-Gene_or_gene_product
.	O

Sunitinib	B-Simple_chemical
is	O
an	O
oral	B-Organism_subdivision
multitargeted	O
receptor	O
tyrosine	O
kinase	O
inhibitor	O
with	O
antiangiogenic	O
and	O
antitumor	B-Cancer
activity	O
that	O
mainly	O
targets	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
receptors	I-Gene_or_gene_product
(	O
VEGFRs	B-Gene_or_gene_product
)	O
.	O

Very	O
recently	O
,	O
sunitinib	B-Simple_chemical
has	O
been	O
shown	O
to	O
be	O
an	O
active	O
agent	O
for	O
the	O
treatment	O
of	O
malignant	B-Cancer
pheochromocytomas	I-Cancer
.	O

However	O
,	O
it	O
is	O
unclear	O
whether	O
sunitinib	B-Simple_chemical
acts	O
only	O
through	O
an	O
antiangiogenic	O
mechanism	O
or	O
whether	O
it	O
may	O
also	O
directly	O
target	O
tumor	B-Cell
cells	I-Cell
.	O

Sunitinib	B-Simple_chemical
markedly	O
induced	O
apoptosis	O
of	O
PC12	B-Cell
cells	I-Cell
in	O
a	O
dose	O
-	O
dependent	O
and	O
time	O
-	O
dependent	O
manner	O
.	O

Furthermore	O
,	O
in	O
support	O
of	O
these	O
findings	O
,	O
we	O
found	O
that	O
sunitinib	B-Simple_chemical
induced	O
a	O
reduction	O
in	O
the	O
expression	O
of	O
the	O
antiapoptotic	O
molecule	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
as	O
well	O
as	O
dephosphorylation	O
of	O
the	O
proapoptotic	O
molecule	O
BAD	B-Gene_or_gene_product
,	O
which	O
results	O
in	O
the	O
activation	O
of	O
BAD	B-Gene_or_gene_product
in	O
these	O
cells	B-Cell
.	O

Consistent	O
with	O
these	O
apoptotic	O
effects	O
,	O
our	O
results	O
showed	O
that	O
sunitinib	B-Simple_chemical
inhibited	O
phosphorylation	O
of	O
Akt	B-Gene_or_gene_product
and	O
mTOR	B-Gene_or_gene_product
and	O
was	O
followed	O
by	O
a	O
reduction	O
of	O
S6K1	B-Gene_or_gene_product
,	O
which	O
is	O
a	O
well	O
-	O
known	O
target	O
of	O
mTOR	B-Gene_or_gene_product
.	O

Knockdown	O
of	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
attenuated	O
the	O
sunitinib	B-Simple_chemical
-	O
induced	O
effects	O
,	O
including	O
apoptosis	O
and	O
inhibition	O
of	O
signaling	O
pathways	O
such	O
as	O
the	O
phosphorylation	O
of	O
Akt	B-Gene_or_gene_product
as	O
well	O
as	O
mTOR	B-Gene_or_gene_product
,	O
and	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
which	O
confirmed	O
that	O
these	O
effects	O
could	O
be	O
mediated	O
by	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
.	O

In	O
addition	O
,	O
silencing	O
of	O
S6K1	B-Gene_or_gene_product
induced	O
apoptosis	O
accompanied	O
by	O
a	O
decrease	O
in	O
the	O
phosphorylation	O
of	O
BAD	B-Gene_or_gene_product
and	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
similar	O
to	O
that	O
observed	O
with	O
sunitinib	B-Simple_chemical
treatment	O
.	O

Thus	O
,	O
these	O
results	O
together	O
suggest	O
that	O
sunitinib	B-Simple_chemical
initially	O
exerts	O
its	O
apoptotic	O
effect	O
through	O
the	O
inhibition	O
of	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
which	O
,	O
when	O
followed	O
by	O
reduction	O
of	O
its	O
downstream	O
effectors	O
,	O
including	O
Akt	B-Gene_or_gene_product
/	O
mTOR	B-Gene_or_gene_product
/	O
S6K1	B-Gene_or_gene_product
,	O
may	O
lead	O
to	O
inhibition	O
of	O
the	O
antiapoptotic	O
molecule	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
activation	O
of	O
the	O
proapoptotic	O
molecule	O
BAD	B-Gene_or_gene_product
in	O
PC12	B-Cell
cells	I-Cell
.	O

However	O
,	O
PC12	B-Cell
cells	I-Cell
do	O
not	O
precisely	O
reflect	O
the	O
pathogenesis	O
of	O
malignant	B-Cell
cells	I-Cell
.	O

Therefore	O
,	O
we	O
confirmed	O
the	O
key	O
findings	O
by	O
replicating	O
these	O
experiments	O
in	O
human	B-Organism
neuroblastoma	B-Cell
SK	I-Cell
-	I-Cell
N	I-Cell
-	I-Cell
SH	I-Cell
cells	I-Cell
.	O

Development	O
of	O
a	O
mouse	B-Organism
model	O
for	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
with	O
endometrial	B-Cancer
cancer	I-Cancer
.	O

Controlling	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
is	O
currently	O
a	O
key	O
issue	O
in	O
cancer	B-Cancer
therapy	O
.	O

Lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
is	O
one	O
of	O
the	O
most	O
important	O
prognostic	O
factors	O
in	O
various	O
types	O
of	O
cancers	B-Cancer
,	O
including	O
endometrial	B-Cancer
cancer	I-Cancer
.	O

Vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
)	O
plays	O
a	O
crucial	O
role	O
in	O
lymphangiogenesis	O
,	O
and	O
is	O
implicated	O
to	O
play	O
an	O
important	O
role	O
in	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
.	O

To	O
evaluate	O
the	O
role	O
of	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
in	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
,	O
we	O
developed	O
an	O
animal	O
model	O
by	O
using	O
an	O
endometrial	B-Cell
cancer	I-Cell
cell	I-Cell
line	I-Cell
,	O
HEC1A	B-Cell
.	O

This	O
cell	B-Cell
line	I-Cell
is	O
not	O
invasive	O
by	O
nature	O
and	O
secretes	O
moderate	O
amounts	O
of	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
;	O
intrauterine	B-Immaterial_anatomical_entity
injection	O
of	O
HEC1A	B-Cell
cells	I-Cell
into	O
Balb	B-Organism
/	I-Organism
c	I-Organism
nude	I-Organism
mice	I-Organism
resulted	O
in	O
uterine	B-Cancer
cancer	I-Cancer
with	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
after	O
8	O
weeks	O
.	O

To	O
analyze	O
the	O
contribution	O
of	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
to	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
,	O
its	O
corresponding	O
gene	O
was	O
stably	O
introduced	O
into	O
HEC1A	B-Cell
cells	I-Cell
(	O
HEC1A	B-Cell
/	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
)	O
,	O
which	O
then	O
produced	O
more	O
than	O
10	O
times	O
the	O
amount	O
of	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
.	O

The	O
number	O
of	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastases	O
was	O
significantly	O
higher	O
in	O
HEC1A	O
/	O
VEGF	B-Gene_or_gene_product
-	O
C	O
cells	O
than	O
in	O
HEC1A	B-Cell
cells	I-Cell
(	O
3	O
.	O
2	O
vs	O
1	O
.	O
1	O
nodes	O
/	O
animal	O
,	O
respectively	O
)	O
.	O

Augmented	O
lymphangiogenesis	O
was	O
observed	O
within	O
tumors	B-Cancer
when	O
HEC1A	O
/	O
VEGF	B-Gene_or_gene_product
-	O
C	O
cells	O
were	O
inoculated	O
.	O

These	O
results	O
indicate	O
that	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
plays	O
a	O
critical	O
role	O
in	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
,	O
in	O
addition	O
to	O
serving	O
as	O
a	O
platform	O
to	O
test	O
the	O
efficacy	O
of	O
various	O
therapeutic	O
modalities	O
against	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
.	O

IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
17F	I-Gene_or_gene_product
deficiency	O
inhibits	O
small	B-Organ
intestinal	I-Organ
tumorigenesis	O
in	O
ApcMin	B-Gene_or_gene_product
/	O
+	O
mice	O
.	O

IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
17	I-Gene_or_gene_product
plays	O
an	O
important	O
role	O
in	O
gut	B-Organism_subdivision
homeostasis	O
.	O

However	O
,	O
the	O
role	O
of	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
17F	I-Gene_or_gene_product
in	O
intestinal	B-Organ
tumorigenesis	O
has	O
not	O
been	O
addressed	O
.	O

Here	O
we	O
demonstrate	O
that	O
ablation	O
of	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
17F	I-Gene_or_gene_product
significantly	O
inhibits	O
spontaneous	O
intestinal	B-Organ
tumorigenesis	O
in	O
the	O
small	B-Organ
intestine	I-Organ
of	O
Apc	B-Gene_or_gene_product
(	O
Min	O
/	O
+	O
)	O
mice	O
.	O

IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
17F	I-Gene_or_gene_product
ablation	O
decreased	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1beta	I-Gene_or_gene_product
and	O
Cox	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
expression	O
as	O
well	O
as	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
17	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
C	I-Gene_or_gene_product
(	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
17RC	I-Gene_or_gene_product
)	O
expression	O
,	O
which	O
were	O
increased	O
in	O
tumors	B-Cancer
from	O
Apc	B-Gene_or_gene_product
(	O
Min	O
/	O
+	O
)	O
mice	O
.	O

Lack	O
of	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
17F	I-Gene_or_gene_product
did	O
not	O
reverse	O
the	O
splenomegaly	B-Pathological_formation
but	O
partially	O
restored	O
thymic	B-Organ
atrophy	O
,	O
suggesting	O
a	O
local	O
effect	O
of	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
17F	I-Gene_or_gene_product
in	O
the	O
intestine	B-Organ
.	O

IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
17F	I-Gene_or_gene_product
deficient	O
Apc	B-Gene_or_gene_product
(	O
Min	O
/	O
+	O
)	O
mice	O
showed	O
a	O
significant	O
decrease	O
in	O
immune	B-Cell
cell	I-Cell
infiltration	O
in	O
the	O
lamina	B-Tissue
propria	I-Tissue
.	O

Interestingly	O
,	O
the	O
expression	O
of	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
17A	I-Gene_or_gene_product
from	O
CD4	B-Gene_or_gene_product
T	I-Gene_or_gene_product
cells	O
in	O
the	O
lamina	B-Tissue
propria	I-Tissue
remains	O
unchanged	O
in	O
the	O
absence	O
of	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
17F	I-Gene_or_gene_product
.	O

Collectively	O
,	O
our	O
results	O
suggest	O
the	O
proinflammatory	O
and	O
essential	O
role	O
of	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
17F	I-Gene_or_gene_product
to	O
develop	O
spontaneous	O
intestinal	B-Organ
tumorigenesis	O
in	O
Apc	B-Gene_or_gene_product
(	O
Min	O
/	O
+	O
)	O
mice	O
in	O
the	O
presence	O
of	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
17A	I-Gene_or_gene_product
.	O

Curcumin	B-Simple_chemical
inhibits	O
metastatic	O
progression	O
of	O
breast	B-Cell
cancer	I-Cell
cell	I-Cell
through	O
suppression	O
of	O
urokinase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
type	I-Gene_or_gene_product
plasminogen	I-Gene_or_gene_product
activator	I-Gene_or_gene_product
by	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappa	I-Gene_or_gene_product
B	I-Gene_or_gene_product
signaling	O
pathways	O
.	O

Curcumin	B-Simple_chemical
(	O
1	B-Simple_chemical
,	I-Simple_chemical
7	I-Simple_chemical
-	I-Simple_chemical
bis	I-Simple_chemical
(	I-Simple_chemical
4	I-Simple_chemical
-	I-Simple_chemical
hydroxy	I-Simple_chemical
-	I-Simple_chemical
3	I-Simple_chemical
-	I-Simple_chemical
methoxyphenyl	I-Simple_chemical
)	I-Simple_chemical
-	I-Simple_chemical
1	I-Simple_chemical
,	I-Simple_chemical
6	I-Simple_chemical
-	I-Simple_chemical
heptadiene	I-Simple_chemical
-	I-Simple_chemical
3	I-Simple_chemical
,	I-Simple_chemical
5	I-Simple_chemical
-	I-Simple_chemical
dione	I-Simple_chemical
)	O
,	O
is	O
extracted	O
from	O
the	O
plant	O
Curcuma	B-Organism
longa	I-Organism
.	O

It	O
was	O
recently	O
reported	O
for	O
its	O
anticancer	B-Cancer
effect	O
on	O
several	O
types	O
of	O
cancer	B-Cell
cells	I-Cell
in	O
vitro	O
however	O
,	O
the	O
molecular	O
mechanisms	O
of	O
this	O
anticancer	B-Cancer
effect	O
are	O
not	O
fully	O
understood	O
.	O

In	O
the	O
present	O
study	O
,	O
we	O
evaluated	O
the	O
effects	O
of	O
curcumin	B-Simple_chemical
on	O
human	B-Organism
mammary	B-Cell
epithelial	I-Cell
carcinoma	I-Cell
MCF	I-Cell
-	I-Cell
7	I-Cell
cells	I-Cell
.	O

Cells	B-Cell
were	O
treated	O
with	O
curcumin	B-Simple_chemical
and	O
examined	O
for	O
cell	B-Cell
viability	O
by	O
MTT	O
assay	O
.	O

The	O
cells	B-Cell
invasion	O
was	O
demonstrated	O
by	O
transwell	O
assay	O
.	O

The	O
binding	O
activity	O
of	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
to	O
DNA	B-Cellular_component
was	O
examined	O
in	O
nuclear	B-Organism_substance
extracts	I-Organism_substance
using	O
Trans	B-Gene_or_gene_product
-	I-Gene_or_gene_product
AM	I-Gene_or_gene_product
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
ELISA	O
kit	O
.	O

Western	O
blot	O
was	O
performed	O
to	O
detect	O
the	O
effect	O
of	O
curcumin	B-Simple_chemical
on	O
the	O
expression	O
of	O
uPA	B-Gene_or_gene_product
.	O

Our	O
results	O
showed	O
that	O
curcumin	B-Simple_chemical
dose	O
-	O
dependently	O
inhibited	O
(	O
P	O
<	O
0	O
.	O
05	O
)	O
the	O
proliferation	O
of	O
MCF	B-Cell
-	I-Cell
7	I-Cell
cells	I-Cell
.	O

Meanwhile	O
,	O
the	O
adhesion	O
and	O
invasion	O
ability	O
of	O
MCF	B-Cell
-	I-Cell
7	I-Cell
cells	I-Cell
were	O
sharply	O
inhibited	O
when	O
treated	O
with	O
different	O
concentrations	O
of	O
curcumin	B-Simple_chemical
.	O

Curcumin	B-Simple_chemical
also	O
significantly	O
decreased	O
(	O
P	O
<	O
0	O
.	O
05	O
)	O
the	O
expression	O
of	O
uPA	B-Gene_or_gene_product
and	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
DNA	B-Cellular_component
binding	O
activity	O
,	O
respectively	O
.	O

It	O
is	O
concluded	O
that	O
curcumin	B-Simple_chemical
inhibits	O
the	O
adhesion	O
and	O
invasion	O
of	O
MCF	B-Cell
-	I-Cell
7	I-Cell
cells	I-Cell
through	O
down	O
-	O
regulating	O
the	O
protein	O
expression	O
of	O
uPA	B-Gene_or_gene_product
via	O
of	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
activation	O
.	O

Accordingly	O
,	O
the	O
therapeutic	O
potential	O
of	O
curcumin	B-Simple_chemical
for	O
breast	B-Cancer
cancer	I-Cancer
deserves	O
further	O
study	O
.	O

c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
MYC	I-Gene_or_gene_product
functions	O
as	O
a	O
molecular	O
switch	O
to	O
alter	O
the	O
response	O
of	O
human	B-Organism
mammary	B-Cell
epithelial	I-Cell
cells	I-Cell
to	O
oncostatin	B-Gene_or_gene_product
M	I-Gene_or_gene_product
.	O

Cytokines	O
play	O
an	O
important	O
role	O
in	O
creating	O
an	O
inflammatory	O
microenvironment	O
,	O
which	O
is	O
now	O
considered	O
a	O
hallmark	O
of	O
cancer	B-Cancer
.	O

Although	O
tumor	B-Cell
cells	I-Cell
can	O
exploit	O
cytokine	O
signaling	O
to	O
promote	O
growth	O
,	O
invasion	O
,	O
and	O
metastasis	O
,	O
the	O
response	O
of	O
normal	O
and	O
premalignant	B-Cell
epithelial	I-Cell
cells	I-Cell
to	O
cytokines	O
present	O
in	O
a	O
developing	O
tumor	B-Cancer
microenvironment	O
remains	O
unclear	O
.	O

Oncostatin	B-Gene_or_gene_product
M	I-Gene_or_gene_product
(	O
OSM	B-Gene_or_gene_product
)	O
,	O
an	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
6	I-Gene_or_gene_product
family	O
cytokine	O
responsible	O
for	O
STAT3	B-Gene_or_gene_product
activation	O
,	O
has	O
been	O
implicated	O
in	O
cancer	B-Cancer
development	O
,	O
progression	O
,	O
invasion	O
,	O
and	O
metastasis	O
.	O

Paradoxically	O
,	O
OSM	B-Gene_or_gene_product
can	O
also	O
suppress	O
the	O
growth	O
of	O
normal	O
cells	B-Cell
and	O
certain	B-Cell
tumor	I-Cell
-	I-Cell
derived	I-Cell
cell	I-Cell
lines	I-Cell
.	O

Using	O
isogenic	O
human	B-Organism
mammary	B-Cell
epithelial	I-Cell
cells	I-Cell
(	O
HMEC	B-Cell
)	O
at	O
different	O
stages	O
of	O
neoplastic	O
transformation	O
,	O
we	O
found	O
that	O
OSM	B-Gene_or_gene_product
signaling	O
suppressed	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
MYC	I-Gene_or_gene_product
expression	O
and	O
engaged	O
a	O
p16	B-Gene_or_gene_product
-	O
and	O
p53	B-Gene_or_gene_product
-	O
independent	O
growth	O
arrest	O
that	O
required	O
STAT3	B-Gene_or_gene_product
activity	O
.	O

Inhibition	O
of	O
STAT3	B-Gene_or_gene_product
activation	O
by	O
expressing	O
a	O
dominant	O
-	O
negative	O
STAT3	B-Gene_or_gene_product
protein	O
or	O
a	O
STAT3	B-Gene_or_gene_product
-	O
shRNA	O
prevented	O
the	O
OSM	B-Gene_or_gene_product
-	O
mediated	O
arrest	O
.	O

In	O
addition	O
,	O
expression	O
of	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
MYC	I-Gene_or_gene_product
from	O
a	O
constitutive	O
promoter	O
also	O
abrogated	O
the	O
STAT3	B-Gene_or_gene_product
-	O
mediated	O
arrest	O
,	O
and	O
strikingly	O
,	O
cooperated	O
with	O
OSM	B-Gene_or_gene_product
to	O
promote	O
anchorage	O
-	O
independent	O
growth	O
(	O
AIG	O
)	O
,	O
a	O
property	O
associated	O
with	O
malignant	O
transformation	O
.	O

Cooperative	O
transformation	O
by	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
MYC	I-Gene_or_gene_product
and	O
OSM	B-Gene_or_gene_product
required	O
PI3K	B-Gene_or_gene_product
and	O
AKT	B-Gene_or_gene_product
signaling	O
,	O
showing	O
the	O
importance	O
of	O
multiple	O
signaling	O
pathways	O
downstream	O
of	O
the	O
OSM	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
in	O
defining	O
the	O
cellular	B-Cell
response	O
to	O
cytokines	O
.	O

These	O
findings	O
identify	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
MYC	I-Gene_or_gene_product
as	O
an	O
important	O
molecular	O
switch	O
that	O
alters	O
the	O
cellular	B-Cell
response	O
to	O
OSM	B-Gene_or_gene_product
-	O
mediated	O
signaling	O
from	O
tumor	B-Cancer
suppressive	O
to	O
tumor	B-Cancer
promoting	O
.	O

PTHrP	B-Gene_or_gene_product
drives	O
breast	B-Cancer
tumor	I-Cancer
initiation	O
,	O
progression	O
,	O
and	O
metastasis	O
in	O
mice	B-Organism
and	O
is	O
a	O
potential	O
therapy	O
target	O
.	O

Parathyroid	B-Gene_or_gene_product
hormone	I-Gene_or_gene_product
-	I-Gene_or_gene_product
related	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
(	O
PTHrP	B-Gene_or_gene_product
)	O
is	O
a	O
secreted	O
factor	O
expressed	O
in	O
almost	O
all	O
normal	O
fetal	B-Tissue
and	O
adult	B-Tissue
tissues	I-Tissue
.	O

It	O
is	O
involved	O
in	O
a	O
wide	O
range	O
of	O
developmental	O
and	O
physiological	O
processes	O
,	O
including	O
serum	B-Organism_substance
calcium	B-Simple_chemical
regulation	O
.	O

PTHrP	B-Gene_or_gene_product
is	O
also	O
associated	O
with	O
the	O
progression	O
of	O
skeletal	B-Organ
metastases	O
,	O
and	O
its	O
dysregulated	O
expression	O
in	O
advanced	O
cancers	B-Cancer
causes	O
malignancy	O
-	O
associated	O
hypercalcemia	O
.	O

Although	O
PTHrP	B-Gene_or_gene_product
is	O
frequently	O
expressed	O
by	O
breast	B-Cancer
tumors	I-Cancer
and	O
other	O
solid	B-Cancer
cancers	I-Cancer
,	O
its	O
effects	O
on	O
tumor	B-Cancer
progression	O
are	O
unclear	O
.	O

Here	O
,	O
we	O
demonstrate	O
in	O
mice	B-Organism
pleiotropic	O
involvement	O
of	O
PTHrP	B-Gene_or_gene_product
in	O
key	O
steps	O
of	O
breast	B-Cancer
cancer	I-Cancer
-	O
it	O
influences	O
the	O
initiation	O
and	O
progression	O
of	O
primary	B-Cancer
tumors	I-Cancer
and	O
metastases	O
.	O

Pthrp	B-Gene_or_gene_product
ablation	O
in	O
the	O
mammary	B-Tissue
epithelium	I-Tissue
of	O
the	O
PyMT	B-Gene_or_gene_product
-	O
MMTV	B-Organism
breast	B-Cancer
cancer	I-Cancer
mouse	B-Organism
model	O
caused	O
a	O
delay	O
in	O
primary	O
tumor	B-Cancer
initiation	O
,	O
inhibited	O
tumor	B-Cancer
progression	O
,	O
and	O
reduced	O
metastasis	O
to	O
distal	O
sites	O
.	O

Mechanistically	O
,	O
it	O
reduced	O
expression	O
of	O
molecular	O
markers	O
of	O
cell	B-Cell
proliferation	O
(	O
Ki67	B-Gene_or_gene_product
)	O
and	O
angiogenesis	O
(	O
factor	B-Gene_or_gene_product
VIII	I-Gene_or_gene_product
)	O
,	O
antiapoptotic	O
factor	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
cell	B-Cell
-	O
cycle	O
progression	O
regulator	O
cyclin	B-Gene_or_gene_product
D1	I-Gene_or_gene_product
,	O
and	O
survival	O
factor	O
AKT1	B-Gene_or_gene_product
.	O

PTHrP	B-Gene_or_gene_product
also	O
influenced	O
expression	O
of	O
the	O
adhesion	O
factor	O
CXCR4	B-Gene_or_gene_product
,	O
and	O
coexpression	O
of	O
PTHrP	B-Gene_or_gene_product
and	O
CXCR4	B-Gene_or_gene_product
was	O
crucial	O
for	O
metastatic	O
spread	O
.	O

Importantly	O
,	O
PTHrP	B-Gene_or_gene_product
-	O
specific	O
neutralizing	O
antibodies	O
slowed	O
the	O
progression	O
and	O
metastasis	O
of	O
human	B-Organism
breast	B-Cancer
cancer	I-Cancer
xenografts	I-Cancer
.	O

Our	O
data	O
identify	O
what	O
we	O
believe	O
to	O
be	O
new	O
functions	O
for	O
PTHrP	B-Gene_or_gene_product
in	O
several	O
key	O
steps	O
of	O
breast	B-Cancer
cancer	I-Cancer
and	O
suggest	O
that	O
PTHrP	B-Gene_or_gene_product
may	O
constitute	O
a	O
novel	O
target	O
for	O
therapeutic	O
intervention	O
.	O

Aquaporins	B-Gene_or_gene_product
in	O
human	B-Organism
breast	B-Cancer
cancer	I-Cancer
:	O
identification	O
and	O
involvement	O
in	O
carcinogenesis	O
of	O
breast	B-Cancer
cancer	I-Cancer
.	O

BACKGROUND	O
AND	O
OBJECTIVES	O
:	O
Aquaporins	B-Gene_or_gene_product
(	O
AQPs	B-Gene_or_gene_product
)	O
play	O
important	O
roles	O
in	O
water	B-Simple_chemical
and	O
glycerol	B-Simple_chemical
transport	O
.	O

Recently	O
,	O
the	O
role	O
of	O
AQPs	B-Gene_or_gene_product
in	O
human	B-Organism
carcinogenesis	O
has	O
become	O
an	O
area	O
of	O
great	O
interest	O
.	O

However	O
,	O
little	O
is	O
known	O
about	O
the	O
function	O
of	O
AQPs	B-Gene_or_gene_product
in	O
human	B-Organism
breast	B-Cancer
cancer	I-Cancer
.	O

The	O
aim	O
of	O
this	O
study	O
was	O
to	O
investigate	O
the	O
expression	O
profile	O
of	O
AQPs	B-Gene_or_gene_product
in	O
human	B-Organism
breast	B-Cancer
cancer	I-Cancer
and	O
its	O
significance	O
.	O

METHODS	O
:	O
In	O
this	O
study	O
,	O
we	O
screened	O
the	O
expression	O
profile	O
of	O
AQP0	B-Gene_or_gene_product
-	I-Gene_or_gene_product
12	I-Gene_or_gene_product
in	O
breast	B-Tissue
cancer	I-Tissue
tissues	I-Tissue
and	O
corresponding	O
normal	B-Tissue
tissues	I-Tissue
by	O
RT	O
-	O
PCR	O
,	O
Western	O
blotting	O
and	O
immunohistochemistry	O
.	O

RESULTS	O
:	O
AQP1	B-Gene_or_gene_product
,	O
3	B-Gene_or_gene_product
-	O
5	B-Gene_or_gene_product
,	O
and	O
10	B-Gene_or_gene_product
-	O
12	B-Gene_or_gene_product
were	O
expressed	O
in	O
human	B-Organism
breast	B-Cancer
cancer	I-Cancer
and	O
/	O
or	O
normal	O
breast	B-Tissue
tissues	I-Tissue
,	O
and	O
AQP1	B-Gene_or_gene_product
and	O
3	B-Gene_or_gene_product
-	O
5	B-Gene_or_gene_product
exhibited	O
differential	O
expression	O
.	O

AQP1	B-Gene_or_gene_product
was	O
expressed	O
in	O
cell	B-Cellular_component
membranes	I-Cellular_component
and	O
its	O
expression	O
was	O
higher	O
in	O
cancer	B-Cancer
than	O
that	O
in	O
normal	B-Tissue
tissues	I-Tissue
.	O

AQP4	B-Gene_or_gene_product
was	O
expressed	O
in	O
the	O
cell	B-Cellular_component
membrane	I-Cellular_component
and	O
cytoplasm	B-Organism_substance
and	O
was	O
detected	O
markedly	O
stronger	O
in	O
normal	O
than	O
in	O
cancer	B-Tissue
tissues	I-Tissue
.	O

AQP5	B-Gene_or_gene_product
was	O
expressed	O
mainly	O
in	O
cell	B-Cellular_component
membranes	I-Cellular_component
in	O
carcinoma	B-Tissue
tissues	I-Tissue
,	O
but	O
was	O
almost	O
absent	O
in	O
normal	O
breast	B-Tissue
tissues	I-Tissue
.	O

Expression	O
of	O
AQP5	B-Gene_or_gene_product
was	O
associated	O
with	O
cellular	B-Cell
differentiation	O
,	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
invasion	O
,	O
and	O
clinicopathological	O
staging	O
.	O

CONCLUSIONS	O
:	O
These	O
observations	O
suggested	O
that	O
several	O
subtypes	O
of	O
the	O
AQP	B-Gene_or_gene_product
family	O
play	O
a	O
role	O
in	O
human	B-Organism
breast	B-Organism_subdivision
carcinogenesis	O
.	O

Research	O
and	O
discovery	O
of	O
the	O
first	O
human	B-Organism
cancer	B-Cancer
virus	O
,	O
HTLV	B-Organism
-	I-Organism
1	I-Organism
.	O

Human	B-Organism
T	I-Organism
-	I-Organism
cell	I-Organism
lymphoma	I-Organism
virus	I-Organism
(	I-Organism
HTLV	I-Organism
)	I-Organism
-	I-Organism
1	I-Organism
was	O
the	O
first	O
human	B-Organism
retrovirus	O
to	O
be	O
discovered	O
.	O

It	O
has	O
been	O
recognized	O
as	O
the	O
cause	O
of	O
adult	B-Cancer
T	I-Cancer
-	I-Cancer
cell	I-Cancer
leukemia	I-Cancer
(	O
ATL	B-Cancer
)	O
.	O

In	O
addition	O
to	O
giving	O
a	O
historical	O
perspective	O
on	O
HTLV	B-Organism
-	I-Organism
1	I-Organism
and	O
other	O
retrovirus	O
research	O
,	O
this	O
paper	O
discusses	O
the	O
origin	O
of	O
HTLV	B-Organism
-	I-Organism
1	I-Organism
;	O
the	O
modes	O
of	O
transmission	O
and	O
global	O
epidemiology	O
of	O
HTLV	B-Organism
-	I-Organism
1	I-Organism
infection	O
;	O
the	O
genome	O
of	O
HTLV	B-Organism
-	I-Organism
1	I-Organism
and	O
the	O
mechanism	O
of	O
HTLV	B-Organism
-	I-Organism
1	I-Organism
-	O
induced	O
leukemogenesis	O
;	O
the	O
role	O
of	O
HTLV	B-Organism
-	I-Organism
1	I-Organism
in	O
other	O
diseases	O
,	O
and	O
recent	O
breakthroughs	O
in	O
ATL	B-Cancer
therapy	O
.	O

Selective	O
inhibition	O
of	O
nuclear	B-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
by	O
nuclear	B-Simple_chemical
factor	I-Simple_chemical
-	I-Simple_chemical
kappaB	I-Simple_chemical
essential	I-Simple_chemical
modulator	I-Simple_chemical
-	I-Simple_chemical
binding	I-Simple_chemical
domain	I-Simple_chemical
peptide	I-Simple_chemical
suppresses	O
the	O
metastasis	O
of	O
highly	O
metastatic	O
oral	B-Cancer
squamous	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
.	O

Nuclear	B-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
(	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
)	O
activation	O
contributes	O
to	O
the	O
development	O
of	O
metastasis	O
,	O
thus	O
leading	O
to	O
a	O
poor	O
prognosis	O
in	O
many	O
cancers	B-Cancer
,	O
including	O
OSCC	B-Cancer
.	O

However	O
,	O
little	O
in	O
vivo	O
experimental	O
data	O
are	O
available	O
about	O
the	O
effects	O
of	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
inhibition	O
on	O
OSCC	B-Cancer
metastasis	O
.	O

OSCC	B-Cell
sublines	I-Cell
were	O
established	O
from	O
a	O
GFP	B-Gene_or_gene_product
-	O
expressing	O
parental	B-Cell
cell	I-Cell
line	I-Cell
,	O
GSAS	B-Cell
,	O
and	O
designated	O
GSAS	B-Cell
/	I-Cell
N3	I-Cell
and	O
N5	B-Cell
according	O
to	O
the	O
in	O
vivo	O
passage	O
number	O
after	O
cervical	B-Multi-tissue_structure
lymph	I-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
by	O
a	O
serial	O
orthotopic	O
transplantation	O
model	O
.	O

In	O
vitro	O
migration	O
and	O
invasion	O
were	O
assessed	O
in	O
these	O
cells	B-Cell
,	O
and	O
the	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
activities	O
and	O
expression	O
of	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
-	O
regulated	O
metastasis	O
-	O
related	O
molecules	O
were	O
also	O
examined	O
.	O

In	O
in	O
vivo	O
experiments	O
,	O
the	O
metastasis	O
and	O
survival	O
of	O
tumor	B-Cancer
-	O
engrafted	O
mice	B-Organism
were	O
monitored	O
.	O

Furthermore	O
,	O
the	O
effects	O
of	O
a	O
selective	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
inhibitor	O
,	O
NEMO	B-Simple_chemical
-	I-Simple_chemical
binding	I-Simple_chemical
domain	I-Simple_chemical
(	I-Simple_chemical
NBD	I-Simple_chemical
)	I-Simple_chemical
peptide	I-Simple_chemical
,	O
on	O
metastasis	O
in	O
GSAS	B-Cell
/	I-Cell
N5	I-Cell
-	O
engrafted	O
mice	B-Organism
were	O
assessed	O
,	O
and	O
engrafted	O
tongue	B-Cancer
tumors	I-Cancer
were	O
immunohistochemically	O
examined	O
.	O

Highly	O
metastatic	O
GSAS	B-Cell
/	I-Cell
N3	I-Cell
and	O
N5	B-Cell
cells	I-Cell
showed	O
an	O
enhanced	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
activity	O
,	O
thus	O
contributing	O
to	O
increased	O
migration	O
,	O
invasion	O
,	O
and	O
a	O
poor	O
prognosis	O
compared	O
with	O
the	O
parent	O
cells	B-Cell
.	O

Furthermore	O
,	O
the	O
expression	O
levels	O
of	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
-	O
regulated	O
metastasis	O
-	O
related	O
molecules	O
,	O
such	O
as	O
fibronectin	B-Gene_or_gene_product
,	O
beta1	B-Gene_or_gene_product
integrin	I-Gene_or_gene_product
,	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
-	B-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
-	B-Gene_or_gene_product
9	I-Gene_or_gene_product
,	O
and	O
-	B-Gene_or_gene_product
14	I-Gene_or_gene_product
,	O
and	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
,	O
were	O
upregulated	O
in	O
the	O
highly	O
metastatic	O
cells	B-Cell
.	O

The	O
NBD	B-Simple_chemical
peptide	I-Simple_chemical
suppressed	O
metastasis	O
and	O
tongue	B-Cancer
tumor	I-Cancer
growth	O
in	O
GSAS	B-Cell
/	I-Cell
N5	I-Cell
-	O
inoculated	O
mice	B-Organism
,	O
and	O
was	O
accompanied	O
by	O
the	O
downregulation	O
of	O
the	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
-	O
regulated	O
metastasis	O
-	O
related	O
molecules	O
in	O
engrafted	O
tongue	B-Cancer
tumors	I-Cancer
.	O

Our	O
results	O
suggest	O
that	O
the	O
selective	O
inhibition	O
of	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
activation	O
by	O
NBD	B-Simple_chemical
peptide	I-Simple_chemical
may	O
provide	O
an	O
effective	O
approach	O
for	O
the	O
treatment	O
of	O
highly	O
metastatic	O
OSCC	B-Cancer
.	O

beta	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
signaling	O
controls	O
metastasis	O
in	O
Braf	B-Gene_or_gene_product
-	O
activated	O
Pten	B-Gene_or_gene_product
-	O
deficient	O
melanomas	B-Cancer
.	O

Malignant	B-Cancer
melanoma	I-Cancer
is	O
characterized	O
by	O
frequent	O
metastasis	O
,	O
however	O
,	O
specific	O
changes	O
that	O
regulate	O
this	O
process	O
have	O
not	O
been	O
clearly	O
delineated	O
.	O

Although	O
it	O
is	O
well	O
known	O
that	O
Wnt	B-Gene_or_gene_product
signaling	O
is	O
frequently	O
dysregulated	O
in	O
melanoma	B-Cancer
,	O
the	O
functional	O
implications	O
of	O
this	O
observation	O
are	O
unclear	O
.	O

By	O
modulating	O
beta	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
levels	O
in	O
a	O
mouse	B-Organism
model	O
of	O
melanoma	B-Cancer
that	O
is	O
based	O
on	O
melanocyte	B-Cell
-	O
specific	O
Pten	B-Gene_or_gene_product
loss	O
and	O
Braf	B-Gene_or_gene_product
(	O
V600E	B-Amino_acid
)	O
mutation	O
,	O
we	O
demonstrate	O
that	O
beta	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
is	O
a	O
central	O
mediator	O
of	O
melanoma	B-Cancer
metastasis	O
to	O
the	O
lymph	B-Multi-tissue_structure
nodes	I-Multi-tissue_structure
and	O
lungs	B-Organ
.	O

In	O
addition	O
to	O
altering	O
metastasis	O
,	O
beta	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
levels	O
control	O
tumor	B-Cancer
differentiation	O
and	O
regulate	O
both	O
MAPK	B-Gene_or_gene_product
/	O
Erk	B-Gene_or_gene_product
and	O
PI3K	B-Gene_or_gene_product
/	O
Akt	B-Gene_or_gene_product
signaling	O
.	O

Highly	O
metastatic	O
tumors	B-Cancer
with	O
beta	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
stabilization	O
are	O
very	O
similar	O
to	O
a	O
subset	O
of	O
human	B-Organism
melanomas	B-Cancer
.	O

Together	O
these	O
findings	O
establish	O
Wnt	B-Gene_or_gene_product
signaling	O
as	O
a	O
metastasis	O
regulator	O
in	O
melanoma	B-Cancer
.	O

Mitochondrial	B-Cellular_component
apoptosis	O
and	O
FAK	B-Gene_or_gene_product
signaling	O
disruption	O
by	O
a	O
novel	O
histone	B-Gene_or_gene_product
deacetylase	I-Gene_or_gene_product
inhibitor	O
,	O
HTPB	B-Simple_chemical
,	O
in	O
antitumor	B-Cancer
and	O
antimetastatic	O
mouse	B-Organism
models	O
.	O

BACKGROUND	O
:	O
Compound	O
targeting	O
histone	B-Gene_or_gene_product
deacetylase	I-Gene_or_gene_product
(	O
HDAC	B-Gene_or_gene_product
)	O
represents	O
a	O
new	O
era	O
in	O
molecular	O
cancer	B-Cancer
therapeutics	O
.	O

However	O
,	O
effective	O
HDAC	B-Gene_or_gene_product
inhibitors	O
for	O
the	O
treatment	O
of	O
solid	B-Cancer
tumors	I-Cancer
remain	O
to	O
be	O
developed	O
.	O

METHODOLOGY	O
/	O
PRINCIPAL	O
FINDINGS	O
:	O
Here	O
,	O
we	O
propose	O
a	O
novel	O
HDAC	B-Gene_or_gene_product
inhibitor	O
,	O
N	B-Simple_chemical
-	I-Simple_chemical
Hydroxy	I-Simple_chemical
-	I-Simple_chemical
4	I-Simple_chemical
-	I-Simple_chemical
(	I-Simple_chemical
4	I-Simple_chemical
-	I-Simple_chemical
phenylbutyryl	I-Simple_chemical
-	I-Simple_chemical
amino	I-Simple_chemical
)	I-Simple_chemical
benzamide	I-Simple_chemical
(	O
HTPB	B-Simple_chemical
)	O
,	O
as	O
a	O
potential	O
chemotherapeutic	O
drug	O
for	O
solid	B-Cancer
tumors	I-Cancer
.	O

The	O
HDAC	B-Gene_or_gene_product
inhibition	O
of	O
HTPB	B-Simple_chemical
was	O
confirmed	O
using	O
HDAC	B-Gene_or_gene_product
activity	O
assay	O
.	O

The	O
antiproliferative	O
and	O
anti	O
-	O
migratory	O
mechanisms	O
of	O
HTPB	B-Simple_chemical
were	O
investigated	O
by	O
cell	B-Cell
proliferation	O
,	O
flow	O
cytometry	O
,	O
DNA	B-Cellular_component
ladder	O
,	O
caspase	B-Gene_or_gene_product
activity	O
,	O
Rho	B-Gene_or_gene_product
activity	O
,	O
F	B-Gene_or_gene_product
-	I-Gene_or_gene_product
actin	I-Gene_or_gene_product
polymerization	O
,	O
and	O
gelatin	O
-	O
zymography	O
for	O
matrix	B-Gene_or_gene_product
metalloproteinases	I-Gene_or_gene_product
(	O
MMPs	B-Gene_or_gene_product
)	O
.	O

Mice	B-Organism
with	O
tumor	B-Cancer
xenograft	I-Cancer
and	O
experimental	O
metastasis	O
model	O
were	O
used	O
to	O
evaluate	O
effects	O
on	O
tumor	B-Cancer
growth	O
and	O
metastasis	O
.	O

Our	O
results	O
indicated	O
that	O
HTPB	B-Simple_chemical
was	O
a	O
pan	O
-	O
HDAC	B-Gene_or_gene_product
inhibitor	O
in	O
suppressing	O
cell	B-Cell
viability	O
specifically	O
of	O
lung	B-Cell
cancer	I-Cell
cells	I-Cell
but	O
not	O
of	O
the	O
normal	B-Cell
lung	I-Cell
cells	I-Cell
.	O

Upon	O
HTPB	B-Simple_chemical
treatment	O
,	O
cell	B-Cell
cycle	O
arrest	O
was	O
induced	O
and	O
subsequently	O
led	O
to	O
mitochondria	B-Cellular_component
-	O
mediated	O
apoptosis	O
.	O

HTPB	B-Simple_chemical
disrupted	O
F	B-Gene_or_gene_product
-	I-Gene_or_gene_product
actin	I-Gene_or_gene_product
dynamics	O
via	O
downregulating	O
RhoA	B-Gene_or_gene_product
activity	O
.	O

Moreover	O
,	O
HTPB	B-Simple_chemical
inhibited	O
activity	O
of	O
MMP2	B-Gene_or_gene_product
and	O
MMP9	B-Gene_or_gene_product
,	O
reduced	O
integrin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta1	I-Gene_or_gene_product
/	O
focal	B-Gene_or_gene_product
adhesion	I-Gene_or_gene_product
complex	I-Gene_or_gene_product
formation	O
and	O
decreased	O
pericellular	O
poly	O
-	O
fibronectin	B-Gene_or_gene_product
assemblies	O
.	O

Finally	O
,	O
intraperitoneal	B-Immaterial_anatomical_entity
injection	O
or	O
oral	B-Organism_subdivision
administration	O
of	O
HTPB	B-Simple_chemical
efficiently	O
inhibited	O
A549	B-Cancer
xenograft	I-Cancer
tumor	I-Cancer
growth	O
in	O
vivo	O
without	O
side	O
effects	O
.	O

HTPB	B-Simple_chemical
delayed	O
lung	B-Organ
metastasis	O
of	O
4T1	O
mouse	B-Organism
breast	O
cancer	O
cells	O
.	O

Acetylation	O
of	O
histone	B-Gene_or_gene_product
and	O
non	O
-	O
histone	B-Gene_or_gene_product
proteins	O
,	O
induction	O
of	O
apoptotic	O
-	O
related	O
proteins	O
and	O
de	O
-	O
phosphorylation	O
of	O
focal	B-Gene_or_gene_product
adhesion	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
were	O
confirmed	O
in	O
treated	O
mice	B-Organism
.	O

CONCLUSIONS	O
/	O
SIGNIFICANCE	O
:	O
These	O
results	O
suggested	O
that	O
intrinsic	O
apoptotic	O
pathway	O
may	O
involve	O
in	O
anti	O
-	O
tumor	B-Cancer
growth	O
effects	O
of	O
HTPB	B-Simple_chemical
in	O
lung	B-Cell
cancer	I-Cell
cells	I-Cell
.	O

HTPB	B-Simple_chemical
significantly	O
suppresses	O
tumor	B-Cancer
metastasis	O
partly	O
through	O
inhibition	O
of	O
integrin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta1	I-Gene_or_gene_product
/	O
FAK	B-Gene_or_gene_product
/	O
MMP	B-Gene_or_gene_product
/	O
RhoA	B-Gene_or_gene_product
/	O
F	B-Gene_or_gene_product
-	I-Gene_or_gene_product
actin	I-Gene_or_gene_product
pathways	O
.	O

We	O
have	O
provided	O
convincing	O
preclinical	O
evidence	O
that	O
HTPB	B-Simple_chemical
is	O
a	O
potent	O
HDAC	B-Gene_or_gene_product
targeted	O
inhibitor	O
and	O
is	O
thus	O
a	O
promising	O
candidate	O
for	O
lung	B-Cancer
cancer	I-Cancer
chemotherapy	O
.	O

Expression	O
of	O
HER2	B-Gene_or_gene_product
and	O
bradykinin	B-Gene_or_gene_product
B1	I-Gene_or_gene_product
receptors	I-Gene_or_gene_product
in	O
precursor	B-Pathological_formation
lesions	I-Pathological_formation
of	O
gallbladder	B-Cancer
carcinoma	I-Cancer
.	O

AIM	O
:	O
To	O
determine	O
the	O
expression	O
of	O
HER2	B-Gene_or_gene_product
and	O
bradykinin	B-Gene_or_gene_product
B	I-Gene_or_gene_product
(	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	I-Gene_or_gene_product
receptors	I-Gene_or_gene_product
(	O
B	B-Gene_or_gene_product
(	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	I-Gene_or_gene_product
R	I-Gene_or_gene_product
)	O
in	O
the	O
two	O
pathogenic	O
models	O
of	O
gallbladder	B-Cancer
cancer	I-Cancer
:	O
the	O
metaplasia	B-Cancer
-	I-Cancer
dysplasia	I-Cancer
-	I-Cancer
carcinoma	I-Cancer
and	O
the	O
adenoma	B-Cancer
-	I-Cancer
carcinoma	I-Cancer
pathways	O
.	O

METHODS	O
:	O
Receptor	O
proteins	O
were	O
visualized	O
by	O
immunohistochemistry	O
on	O
5	O
-	O
mum	O
sections	B-Multi-tissue_structure
of	O
paraffin	O
-	O
embedded	O
tissue	B-Tissue
.	O

Expression	O
of	O
both	O
receptors	O
was	O
studied	O
in	O
biopsy	B-Cancer
samples	I-Cancer
from	O
92	O
patients	B-Organism
(	O
6	O
males	O
and	O
86	O
females	O
;	O
age	O
ranging	O
from	O
28	O
to	O
86	O
years	O
,	O
mean	O
56	O
years	O
)	O
.	O

High	O
HER2	B-Gene_or_gene_product
expression	O
in	O
specimens	B-Cancer
was	O
additionally	O
investigated	O
by	O
fluorescence	O
in	O
situ	O
hybridization	O
.	O

Cell	B-Cell
proliferation	O
in	O
each	O
sample	O
was	O
assessed	O
by	O
using	O
the	O
Ki	O
-	O
67	O
proliferation	O
marker	O
.	O

RESULTS	O
:	O
HER2	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
protein	O
was	O
absent	O
in	O
adenomas	B-Cancer
and	O
in	O
normal	O
gallbladder	B-Tissue
epithelium	I-Tissue
.	O

On	O
the	O
contrary	O
,	O
there	O
was	O
intense	O
staining	O
for	O
HER2	B-Gene_or_gene_product
on	O
the	O
basolateral	B-Cellular_component
membrane	I-Cellular_component
of	O
epithelial	B-Cell
cells	I-Cell
of	O
intestinal	B-Cancer
metaplasia	I-Cancer
(	O
22	O
/	O
24	O
;	O
91	O
.	O
7	O
%	O
)	O
and	O
carcinoma	B-Cancer
in	O
situ	O
(	O
9	O
/	O
10	O
;	O
90	O
%	O
)	O
,	O
the	O
lesions	B-Pathological_formation
that	O
displayed	O
a	O
significantly	O
high	O
proliferation	O
index	O
.	O

Protein	O
up	O
-	O
regulation	O
of	O
HER2	B-Gene_or_gene_product
in	O
the	O
epithelium	B-Tissue
with	O
metaplasia	B-Cancer
or	O
carcinoma	B-Cancer
in	O
situ	O
was	O
not	O
accompanied	O
by	O
HER2	B-Gene_or_gene_product
gene	O
amplification	O
.	O

A	O
similar	O
result	O
was	O
observed	O
in	O
invasive	B-Cancer
carcinomas	I-Cancer
(	O
0	O
/	O
12	O
)	O
.	O

The	O
B	B-Gene_or_gene_product
(	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	I-Gene_or_gene_product
R	I-Gene_or_gene_product
distribution	O
pattern	O
mirrored	O
that	O
of	O
HER2	B-Gene_or_gene_product
except	O
that	O
B	B-Gene_or_gene_product
(	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	I-Gene_or_gene_product
R	I-Gene_or_gene_product
was	O
additionally	O
observed	O
in	O
the	O
adenomas	B-Cancer
.	O

The	O
B	B-Gene_or_gene_product
(	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	I-Gene_or_gene_product
R	I-Gene_or_gene_product
appeared	O
either	O
as	O
cytoplasmic	B-Organism_substance
dots	O
or	O
labeling	O
on	O
the	O
apical	B-Cellular_component
cell	I-Cellular_component
membrane	I-Cellular_component
of	O
the	O
cells	B-Cell
composing	O
the	O
epithelia	B-Tissue
with	O
intestinal	B-Cancer
metaplasia	I-Cancer
(	O
24	O
/	O
24	O
;	O
100	O
%	O
)	O
and	O
carcinoma	B-Cancer
in	I-Cancer
situ	I-Cancer
(	O
10	O
/	O
10	O
;	O
100	O
%	O
)	O
and	O
in	O
the	O
epithelial	B-Cell
cells	I-Cell
of	O
adenomas	B-Cancer
.	O

In	O
contrast	O
,	O
both	O
HER2	B-Gene_or_gene_product
(	O
4	O
/	O
12	O
;	O
33	O
%	O
)	O
and	O
B	B-Gene_or_gene_product
(	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	I-Gene_or_gene_product
R	I-Gene_or_gene_product
(	O
1	O
/	O
12	O
;	O
8	O
.	O
3	O
%	O
)	O
showed	O
a	O
low	O
expression	O
in	O
invasive	B-Cancer
gallbladder	I-Cancer
carcinomas	I-Cancer
.	O

CONCLUSION	O
:	O
The	O
up	O
-	O
regulation	O
of	O
HER2	B-Gene_or_gene_product
and	O
B	B-Gene_or_gene_product
(	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	I-Gene_or_gene_product
R	I-Gene_or_gene_product
in	O
precursor	B-Pathological_formation
lesions	I-Pathological_formation
of	O
gallbladder	B-Cancer
carcinoma	I-Cancer
suggests	O
cross	O
-	O
talk	O
between	O
these	O
two	O
receptors	O
that	O
may	O
be	O
of	O
importance	O
in	O
the	O
modulation	O
of	O
cell	B-Cell
proliferation	O
in	O
gallbladder	B-Organ
carcinogenesis	O
.	O

Expression	O
of	O
basic	B-Gene_or_gene_product
fibroblast	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
in	O
normal	O
human	B-Organism
tissues	B-Tissue
.	O

The	O
distribution	O
of	O
basic	B-Gene_or_gene_product
fibroblast	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
bFGF	B-Gene_or_gene_product
)	O
was	O
studied	O
immunohistochemically	O
in	O
fresh	O
frozen	O
sections	O
of	O
normal	O
human	B-Organism
tissues	B-Tissue
.	O

Immunodetection	O
was	O
performed	O
with	O
a	O
specific	O
anti	O
-	O
bFGF	B-Gene_or_gene_product
mouse	B-Organism
monoclonal	O
antibody	O
that	O
was	O
found	O
to	O
react	O
with	O
recombinant	O
human	B-Organism
bFGF	B-Gene_or_gene_product
in	O
Western	O
blot	O
analysis	O
,	O
and	O
to	O
specifically	O
neutralize	O
the	O
mitogenic	O
activity	O
of	O
bFGF	B-Gene_or_gene_product
on	O
bovine	B-Organism
vascular	B-Cell
endothelial	I-Cell
cells	I-Cell
.	O

Expression	O
of	O
bFGF	B-Gene_or_gene_product
on	O
normal	O
human	B-Organism
tissues	B-Tissue
was	O
ubiquitously	O
detected	O
in	O
the	O
basement	B-Cellular_component
membranes	I-Cellular_component
of	O
all	O
size	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
,	O
but	O
was	O
not	O
found	O
in	O
epidermal	B-Cellular_component
or	O
epithelial	B-Cellular_component
basement	I-Cellular_component
membranes	I-Cellular_component
of	O
a	O
variety	O
of	O
tissues	B-Tissue
tested	O
.	O

Intensity	O
and	O
patterns	O
of	O
localization	O
in	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
was	O
consistent	O
in	O
various	O
tissues	B-Tissue
,	O
but	O
varied	O
among	O
different	O
regions	O
of	O
the	O
vascular	B-Multi-tissue_structure
bed	I-Multi-tissue_structure
.	O

Whereas	O
homogeneous	O
and	O
intense	O
immunoreactivity	O
were	O
observed	O
in	O
large	O
and	O
intermediate	O
size	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
,	O
heterogeneity	O
of	O
expression	O
was	O
found	O
in	O
capillaries	B-Tissue
.	O

The	O
most	O
intense	O
immunoreactivity	O
was	O
observed	O
in	O
branching	O
capillaries	B-Tissue
.	O

Endothelial	B-Cell
cell	I-Cell
staining	O
was	O
heterogeneous	O
and	O
varied	O
in	O
different	O
regions	O
.	O

Strong	O
staining	O
for	O
bFGF	B-Gene_or_gene_product
was	O
also	O
found	O
in	O
cardiac	B-Cell
muscle	I-Cell
fibers	I-Cell
,	O
smooth	B-Cell
muscle	I-Cell
cells	I-Cell
of	O
mid	O
-	O
size	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
,	O
the	O
gut	B-Organism_subdivision
and	O
the	O
myometrium	B-Multi-tissue_structure
,	O
in	O
central	B-Cell
nervous	I-Cell
system	I-Cell
neurons	I-Cell
and	O
cerebellar	B-Cell
Purkinje	I-Cell
cells	I-Cell
,	O
and	O
on	O
epithelial	B-Cell
cells	I-Cell
of	O
the	O
bronchi	B-Multi-tissue_structure
,	O
colon	B-Multi-tissue_structure
,	O
endometrium	B-Multi-tissue_structure
,	O
and	O
sweat	B-Multi-tissue_structure
gland	I-Multi-tissue_structure
ducts	I-Multi-tissue_structure
of	O
the	O
skin	B-Organ
.	O

The	O
presence	O
of	O
bFGF	B-Gene_or_gene_product
in	O
the	O
extracellular	B-Immaterial_anatomical_entity
compartment	I-Immaterial_anatomical_entity
of	O
a	O
diverse	O
variety	O
of	O
organs	B-Organ
may	O
play	O
a	O
role	O
in	O
angiogenesis	O
.	O

However	O
,	O
the	O
function	O
of	O
bFGF	B-Gene_or_gene_product
in	O
parenchymal	B-Cell
cells	I-Cell
remains	O
to	O
be	O
determined	O
.	O

Overexpression	O
confers	O
an	O
oncogenic	O
potential	O
upon	O
the	O
eph	B-Gene_or_gene_product
gene	O
.	O

The	O
eph	B-Gene_or_gene_product
gene	O
encodes	O
a	O
putative	O
receptor	B-Gene_or_gene_product
tyrosine	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
for	O
an	O
as	O
yet	O
unknown	O
ligand	O
.	O

Some	O
human	B-Organism
cancer	B-Cell
cells	I-Cell
have	O
been	O
found	O
to	O
overexpress	O
eph	B-Gene_or_gene_product
mRNAs	O
without	O
gene	O
amplification	O
.	O

We	O
show	O
here	O
that	O
NIH3T3	B-Cell
cells	I-Cell
acquire	O
tumorigenic	O
ability	O
in	O
nude	B-Organism
mice	I-Organism
and	O
make	O
colonies	O
in	O
soft	O
agar	O
with	O
a	O
viral	O
LTR	O
(	O
Long	O
Terminal	O
Repeat	O
)	O
-	O
driven	O
artificial	O
expression	O
of	O
the	O
eph	B-Gene_or_gene_product
gene	O
to	O
a	O
high	O
level	O
.	O

This	O
result	O
supports	O
the	O
alleged	O
contribution	O
of	O
overexpressed	O
receptor	B-Gene_or_gene_product
tyrosine	I-Gene_or_gene_product
kinases	I-Gene_or_gene_product
to	O
cell	B-Cell
transformation	O
.	O

Energy	O
supply	O
of	O
the	O
mitotic	O
cell	O
cycle	O
and	O
the	O
Na	B-Simple_chemical
+	I-Simple_chemical
/	O
H	B-Simple_chemical
+	I-Simple_chemical
-	O
antiport	O
in	O
ascites	B-Cancer
tumors	I-Cancer
.	O

The	O
activation	O
of	O
Na	B-Simple_chemical
+	I-Simple_chemical
transport	O
is	O
due	O
to	O
the	O
exchange	O
of	O
protons	B-Simple_chemical
formed	O
via	O
glucose	B-Simple_chemical
conversion	O
into	O
lactate	B-Simple_chemical
for	O
Na	B-Simple_chemical
+	I-Simple_chemical
,	O
i	O
.	O
e	O
.	O
,	O
to	O
the	O
stimulation	O
of	O
the	O
Na	B-Simple_chemical
+	I-Simple_chemical
/	O
H	B-Simple_chemical
+	I-Simple_chemical
-	O
antiport	O
.	O

Experimental	O
results	O
and	O
theoretical	O
calculations	O
suggest	O
that	O
in	O
glucose	B-Simple_chemical
-	O
containing	O
medium	O
the	O
Na	B-Simple_chemical
+	I-Simple_chemical
transport	O
increases	O
from	O
0	O
.	O
75	O
to	O
1	O
.	O
78	O
pmol	O
/	O
hour	O
per	O
cell	B-Cell
.	O

The	O
permeability	O
of	O
plasma	B-Cellular_component
membranes	I-Cellular_component
for	O
K	B-Simple_chemical
+	I-Simple_chemical
increases	O
2	O
.	O
75	O
fold	O
,	O
while	O
the	O
passive	O
flux	O
of	O
Na	B-Simple_chemical
+	I-Simple_chemical
diminishes	O
.	O

The	O
intensity	O
of	O
O2	B-Simple_chemical
adsorption	O
by	O
ascites	O
tumor	B-Cell
cells	I-Cell
does	O
not	O
practically	O
depend	O
on	O
the	O
monovalent	O
cation	B-Simple_chemical
concentration	O
gradient	O
between	O
the	O
cells	B-Cell
and	O
the	O
culture	O
medium	O
,	O
whereas	O
the	O
rate	O
of	O
glycolysis	O
decreases	O
simultaneously	O
with	O
the	O
diminution	O
of	O
the	O
concentration	O
gradient	O
.	O

In	O
synchronized	O
cultures	B-Cell
at	O
the	O
beginning	O
of	O
the	O
mitotic	O
cycle	O
,	O
the	O
bulk	O
of	O
ATP	B-Simple_chemical
resynthesized	O
via	O
glycolysis	O
is	O
utilized	O
for	O
the	O
synthesis	O
of	O
biopolymers	O
,	O
whereas	O
that	O
at	O
the	O
end	O
of	O
the	O
S	O
-	O
phase	O
and	O
in	O
the	O
G2	O
-	O
phase	O
is	O
utilized	O
for	O
cation	B-Simple_chemical
transport	O
across	O
plasma	B-Cellular_component
membranes	I-Cellular_component
.	O

From	O
35	O
to	O
100	O
%	O
of	O
the	O
whole	O
amount	O
of	O
ATP	B-Simple_chemical
resynthesized	O
via	O
glycolysis	O
is	O
utilized	O
for	O
transport	O
purposes	O
.	O

It	O
is	O
concluded	O
that	O
the	O
observed	O
increase	O
in	O
the	O
Na	B-Simple_chemical
+	I-Simple_chemical
/	O
K	B-Simple_chemical
+	I-Simple_chemical
ratio	O
in	O
ascites	O
tumor	B-Cell
cells	I-Cell
is	O
connected	O
with	O
their	O
enhanced	O
ability	O
to	O
synthesize	O
lactic	B-Simple_chemical
acid	I-Simple_chemical
.	O

Presumably	O
,	O
glycolysis	O
is	O
one	O
of	O
the	O
regulatory	O
mechanisms	O
of	O
intracellular	B-Immaterial_anatomical_entity
ratios	O
of	O
monovalent	O
cations	B-Simple_chemical
.	O

[	O
Tumor	B-Cancer
metastasis	O
and	O
the	O
fibrinolytic	O
system	O
]	O
.	O

Metastatic	O
spread	O
of	O
malignant	B-Cancer
tumor	I-Cancer
appears	O
to	O
correlate	O
with	O
activation	O
of	O
the	O
fibrolytic	O
system	O
.	O

The	O
role	O
of	O
fibrinolysis	O
in	O
growth	O
and	O
metastasis	O
was	O
examined	O
in	O
Lewis	B-Cancer
lung	I-Cancer
carcinoma	I-Cancer
of	O
mice	B-Organism
.	O

The	O
inhibition	O
of	O
fibrinolysis	O
or	O
proteases	O
decreased	O
the	O
primary	O
tumor	B-Cancer
growth	O
and	O
pulmonary	B-Organ
metastasis	O
,	O
whereas	O
the	O
activation	O
of	O
fibrinolysis	O
or	O
proteases	O
increased	O
the	O
number	O
of	O
metastatic	O
foci	O
in	O
the	O
lung	B-Organ
.	O

Electronmicroscopically	O
,	O
thrombus	B-Pathological_formation
formation	O
in	O
the	O
primary	O
site	O
prevented	O
tumor	B-Cancer
invasion	O
and	O
metastasis	O
formation	O
.	O

Plasminogen	B-Gene_or_gene_product
activator	I-Gene_or_gene_product
(	O
PA	B-Gene_or_gene_product
)	O
content	O
of	O
excised	O
tumors	B-Cancer
was	O
determined	O
by	O
SDS	O
-	O
PAGE	O
,	O
and	O
major	O
PA	B-Gene_or_gene_product
was	O
found	O
to	O
be	O
urokinase	B-Gene_or_gene_product
(	O
UK	B-Gene_or_gene_product
)	O
type	O
.	O

Immunohistochemical	O
study	O
with	O
specific	O
antisera	O
was	O
done	O
.	O

When	O
tumor	B-Cell
cells	I-Cell
possessed	O
a	O
high	O
level	O
of	O
UK	B-Gene_or_gene_product
,	O
laminin	B-Gene_or_gene_product
and	O
type	B-Gene_or_gene_product
IV	I-Gene_or_gene_product
collagen	I-Gene_or_gene_product
,	O
components	O
of	O
the	O
basement	B-Cellular_component
membrane	I-Cellular_component
,	O
disappeared	O
from	O
tumor	B-Tissue
tissues	I-Tissue
.	O

These	O
findings	O
suggest	O
that	O
PA	B-Gene_or_gene_product
through	O
protease	O
cascade	O
plays	O
a	O
role	O
in	O
tumor	B-Cancer
invasion	O
and	O
metastasis	O
.	O

Clinically	O
,	O
patients	B-Organism
with	O
advanced	B-Cancer
cancer	I-Cancer
are	O
usually	O
in	O
a	O
hypercoagulable	O
state	O
with	O
elevated	O
fibrinogen	B-Gene_or_gene_product
,	O
and	O
fibrin	B-Gene_or_gene_product
deposition	O
around	O
tumor	B-Cancer
mass	O
is	O
a	O
serious	O
problem	O
in	O
cancer	B-Cancer
chemotherapy	O
.	O

UK	B-Gene_or_gene_product
infusion	O
prior	O
to	O
5	B-Simple_chemical
-	I-Simple_chemical
fluorouracil	I-Simple_chemical
increased	O
tissue	B-Tissue
concentration	O
of	O
antitumor	B-Cancer
agent	O
.	O

However	O
,	O
development	O
of	O
consumption	O
coagulopathy	O
characterized	O
by	O
progression	O
from	O
hypercoagulable	O
state	O
to	O
disseminated	O
intravascular	B-Immaterial_anatomical_entity
coagulation	O
has	O
also	O
been	O
found	O
in	O
several	O
cases	O
.	O

On	O
the	O
mechanism	O
of	O
glycolysis	O
stimulation	O
by	O
neutral	O
detergents	O
in	O
3T3	B-Cell
and	O
Ehrlich	B-Cell
ascites	I-Cell
tumor	I-Cell
cells	I-Cell
.	O

Glycolysis	O
of	O
3T3	B-Cell
and	O
Ehrlich	B-Cell
ascites	I-Cell
tumor	I-Cell
cells	I-Cell
was	O
greatly	O
enhanced	O
by	O
Nonidet	B-Simple_chemical
P	I-Simple_chemical
-	I-Simple_chemical
40	I-Simple_chemical
or	O
Triton	B-Simple_chemical
X	I-Simple_chemical
-	I-Simple_chemical
100	I-Simple_chemical
at	O
about	O
100	O
micrograms	O
/	O
mg	O
cell	B-Cell
protein	O
.	O

This	O
enhanced	O
glycolysis	O
was	O
partly	O
sensitive	O
to	O
rutamycin	B-Simple_chemical
and	O
partly	O
sensitive	O
to	O
ouabain	B-Simple_chemical
,	O
suggesting	O
that	O
the	O
detergent	O
released	O
the	O
control	O
of	O
the	O
ATPase	B-Gene_or_gene_product
of	O
the	O
mitochondria	B-Cellular_component
and	O
of	O
the	O
plasma	B-Cellular_component
membrane	I-Cellular_component
Na	B-Gene_or_gene_product
+	I-Gene_or_gene_product
K	I-Gene_or_gene_product
+	I-Gene_or_gene_product
-	I-Gene_or_gene_product
ATPase	I-Gene_or_gene_product
.	O

Nonidet	B-Simple_chemical
P	I-Simple_chemical
-	I-Simple_chemical
40	I-Simple_chemical
had	O
no	O
effect	O
on	O
glycolysis	O
in	O
cell	B-Organism_substance
-	I-Organism_substance
free	I-Organism_substance
extracts	I-Organism_substance
from	O
Ehrlich	B-Cell
ascites	I-Cell
tumor	I-Cell
cells	I-Cell
to	O
which	O
soluble	O
mitochondrial	B-Cellular_component
ATPase	B-Gene_or_gene_product
was	O
added	O
.	O

Measuring	O
ouabain	B-Simple_chemical
-	O
sensitive	O
22Na	O
efflux	O
and	O
using	O
ouabain	B-Simple_chemical
-	O
sensitive	O
lactate	B-Simple_chemical
production	O
as	O
a	O
measure	O
of	O
ATP	B-Simple_chemical
hydrolysis	O
by	O
the	O
Na	B-Gene_or_gene_product
+	I-Gene_or_gene_product
K	I-Gene_or_gene_product
+	I-Gene_or_gene_product
pump	I-Gene_or_gene_product
,	O
it	O
was	O
shown	O
that	O
Nonidet	B-Simple_chemical
P	I-Simple_chemical
-	I-Simple_chemical
40	I-Simple_chemical
greatly	O
decreased	O
the	O
efficiency	O
of	O
the	O
Na	B-Gene_or_gene_product
+	I-Gene_or_gene_product
K	I-Gene_or_gene_product
+	I-Gene_or_gene_product
pump	I-Gene_or_gene_product
.	O

Quercetin	B-Simple_chemical
increased	O
the	O
efficiency	O
of	O
pumping	O
in	O
EAT	B-Cell
cells	I-Cell
both	O
in	O
the	O
absence	O
and	O
presence	O
of	O
the	O
detergent	O
.	O

Elevated	O
levels	O
of	O
diacylglycerol	B-Simple_chemical
and	O
decreased	O
phorbol	B-Simple_chemical
ester	I-Simple_chemical
sensitivity	O
in	O
ras	B-Gene_or_gene_product
-	O
transformed	O
fibroblasts	B-Cell
.	O

Diacylglycerol	B-Simple_chemical
(	O
DG	B-Simple_chemical
)	O
plays	O
a	O
central	O
role	O
in	O
phospholipid	B-Simple_chemical
metabolism	O
and	O
is	O
an	O
endogenous	O
activator	O
of	O
protein	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
C	I-Gene_or_gene_product
.	O

We	O
have	O
suggested	O
that	O
constitutive	O
activation	O
of	O
this	O
kinase	O
is	O
one	O
mechanism	O
by	O
which	O
oncogenes	O
transform	O
cells	B-Cell
.	O

The	O
ras	B-Gene_or_gene_product
-	O
encoded	O
proteins	O
are	O
similar	O
to	O
regulatory	O
G	B-Gene_or_gene_product
-	I-Gene_or_gene_product
proteins	I-Gene_or_gene_product
and	O
are	O
candidates	O
for	O
the	O
unknown	O
G	B-Gene_or_gene_product
-	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
that	O
modulates	O
phosphatidylinositol	B-Simple_chemical
(	O
PI	B-Simple_chemical
)	O
turnover	O
.	O

Differences	O
in	O
polyphosphoinositide	B-Simple_chemical
metabolism	O
have	O
been	O
reported	O
for	O
ras	B-Gene_or_gene_product
-	O
transformed	O
cells	B-Cell
.	O

But	O
because	O
these	O
experiments	O
were	O
performed	O
on	O
confluent	O
cultures	O
of	O
established	O
cell	B-Cell
lines	I-Cell
,	O
the	O
differences	O
are	O
difficult	O
to	O
attribute	O
to	O
ras	B-Gene_or_gene_product
transformation	O
.	O

Here	O
we	O
show	O
that	O
exponentially	O
growing	O
NIH	B-Cell
3T3	I-Cell
fibroblasts	I-Cell
recently	O
transformed	O
by	O
Ha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
or	O
Ki	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
possess	O
elevated	O
DG	B-Simple_chemical
concentrations	O
without	O
significant	O
alterations	O
in	O
the	O
levels	O
of	O
other	O
polyphosphoinositide	B-Simple_chemical
metabolites	O
.	O

The	O
basal	O
phosphorylation	O
of	O
protein	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
C	I-Gene_or_gene_product
substrate	O
of	O
relative	O
molecular	O
mass	O
(	O
Mr	O
)	O
80	O
,	O
000	O
(	O
80K	O
)	O
is	O
significantly	O
increased	O
in	O
all	O
the	O
ras	B-Gene_or_gene_product
-	O
transformed	O
cell	B-Cell
lines	I-Cell
.	O

Surprisingly	O
,	O
however	O
,	O
further	O
phosphorylation	O
of	O
this	O
protein	O
on	O
addition	O
of	O
phorbol	B-Simple_chemical
ester	I-Simple_chemical
was	O
greatly	O
reduced	O
.	O

Ha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
cells	O
also	O
show	O
less	O
binding	O
of	O
phorbol	B-Simple_chemical
ester	I-Simple_chemical
than	O
control	O
cells	B-Cell
,	O
suggesting	O
that	O
elevation	O
of	O
DG	B-Simple_chemical
causes	O
partial	O
down	O
-	O
regulation	O
in	O
addition	O
to	O
activation	O
of	O
protein	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
C	I-Gene_or_gene_product
.	O

Epidermal	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
and	O
its	O
receptor	O
.	O

Epidermal	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
EGF	B-Gene_or_gene_product
)	O
binds	O
with	O
high	O
affinity	O
and	O
specificity	O
to	O
a	O
single	O
site	O
on	O
the	O
external	O
domain	O
of	O
its	O
transmembrane	B-Cellular_component
receptor	O
to	O
activate	O
the	O
tyrosine	O
protein	O
kinase	O
activity	O
of	O
its	O
cytoplasmic	B-Organism_substance
portion	O
.	O

The	O
EGF	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
gene	O
is	O
amplified	O
and	O
over	O
-	O
expressed	O
in	O
several	O
human	B-Organism
tumors	B-Cancer
,	O
suggesting	O
that	O
increased	O
concentrations	O
of	O
the	O
proto	O
-	O
oncogene	O
leads	O
to	O
constitutive	O
activity	O
similar	O
to	O
that	O
seen	O
with	O
oncogene	O
erb	B-Gene_or_gene_product
B	I-Gene_or_gene_product
.	O

Synthesis	O
and	O
degradation	O
of	O
the	O
EGF	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
are	O
regulated	O
,	O
in	O
addition	O
,	O
covalent	O
modification	O
by	O
phosphorylation	O
regulates	O
activity	O
of	O
the	O
receptor	O
protein	O
.	O

Intramolecular	O
self	O
-	O
phosphorylation	O
of	O
Tyr1173	B-Amino_acid
removes	O
a	O
competitive	O
inhibitory	O
constraint	O
to	O
enhance	O
phosphorylation	O
of	O
substrates	O
.	O

Phosphorylation	O
of	O
Thr654	B-Amino_acid
by	O
protein	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
C	I-Gene_or_gene_product
decreases	O
high	O
affinity	O
EGF	B-Gene_or_gene_product
binding	O
and	O
EGF	B-Gene_or_gene_product
-	O
stimulated	O
tyrosine	O
protein	O
kinase	O
activity	O
,	O
providing	O
a	O
mechanism	O
for	O
heterologous	O
regulation	O
of	O
the	O
EGF	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
by	O
tumor	B-Cancer
promoters	O
and	O
other	O
ligand	B-Gene_or_gene_product
X	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
complexes	O
.	O

Extensive	O
regulation	O
contributes	O
to	O
normal	O
growth	O
control	O
,	O
abrogation	O
of	O
regulatory	O
controls	O
contributes	O
to	O
uncontrolled	O
growth	O
as	O
seen	O
with	O
erb	B-Gene_or_gene_product
B	I-Gene_or_gene_product
transformation	O
and	O
EGF	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
gene	O
amplification	O
in	O
human	B-Organism
tumors	B-Cancer
.	O

Glutamine	B-Amino_acid
and	O
glucose	B-Simple_chemical
as	O
energy	O
substrates	O
for	O
Ehrlich	B-Cell
ascites	I-Cell
tumour	I-Cell
cells	I-Cell
.	O

Energy	O
metabolism	O
of	O
freshly	O
harvested	O
Ehrlich	B-Cell
ascites	I-Cell
tumour	I-Cell
cells	I-Cell
in	O
the	O
presence	O
of	O
5	O
mM	O
glucose	B-Simple_chemical
and	O
/	O
or	O
0	O
.	O
5	O
mM	O
glutamine	B-Amino_acid
was	O
studied	O
.	O

The	O
rate	O
of	O
oxygen	B-Simple_chemical
utilization	O
was	O
not	O
altered	O
by	O
the	O
addition	O
of	O
0	O
.	O
5	O
mM	O
glutamine	B-Amino_acid
;	O
5	O
mM	O
glucose	B-Simple_chemical
induced	O
an	O
inhibition	O
of	O
respiration	O
.	O

In	O
the	O
presence	O
of	O
both	O
glucose	B-Simple_chemical
and	O
glutamine	B-Amino_acid
,	O
the	O
Crabtree	O
effect	O
decreased	O
.	O

In	O
these	O
conditions	O
,	O
the	O
rates	O
of	O
oxygen	B-Simple_chemical
uptake	O
,	O
the	O
CO2	B-Simple_chemical
evolution	O
and	O
the	O
changes	O
in	O
the	O
redox	O
states	O
of	O
cytochromes	B-Gene_or_gene_product
indicate	O
that	O
glucose	B-Simple_chemical
is	O
preferred	O
by	O
Ehrlich	B-Cell
ascites	I-Cell
tumour	I-Cell
cells	I-Cell
as	O
energy	O
substrate	O
.	O

Glucose	B-Simple_chemical
decreased	O
the	O
rate	O
of	O
glutamine	B-Amino_acid
utilization	O
by	O
34	O
%	O
.	O

On	O
the	O
other	O
hand	O
,	O
glutaminolysis	O
did	O
not	O
inhibit	O
glycolysis	O
.	O

Growth	O
state	O
-	O
dependent	O
regulation	O
of	O
protein	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
C	I-Gene_or_gene_product
in	O
normal	O
and	O
transformed	O
murine	B-Organism
cells	B-Cell
.	O

We	O
determined	O
whether	O
growth	O
state	O
can	O
influence	O
the	O
action	O
of	O
protein	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
C	I-Gene_or_gene_product
by	O
measuring	O
protein	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
C	I-Gene_or_gene_product
activity	O
in	O
growing	O
and	O
stationary	O
cultures	O
of	O
normal	O
and	O
transformed	O
cells	B-Cell
.	O

Two	O
approaches	O
were	O
used	O
to	O
measure	O
protein	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
C	I-Gene_or_gene_product
:	O
assay	O
of	O
intact	O
cells	B-Cell
for	O
inhibition	O
of	O
epidermal	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
EGF	B-Gene_or_gene_product
)	O
binding	O
in	O
response	O
to	O
phorbol	B-Simple_chemical
dibutyrate	I-Simple_chemical
(	O
PDBu	B-Simple_chemical
)	O
;	O
and	O
assay	O
of	O
detergent	O
extracts	B-Organism_substance
for	O
total	O
calcium	B-Simple_chemical
,	O
phospholipid	B-Simple_chemical
-	O
dependent	O
kinase	O
activity	O
.	O

In	O
extracts	B-Organism_substance
of	O
growing	O
and	O
stationary	O
Swiss	B-Cell
3T3	I-Cell
cells	I-Cell
,	O
the	O
total	O
amount	O
of	O
protein	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
C	I-Gene_or_gene_product
activity	O
was	O
similar	O
,	O
indicating	O
that	O
growth	O
state	O
does	O
not	O
alter	O
the	O
level	O
of	O
enzyme	O
in	O
the	O
cell	B-Cell
.	O

The	O
short	O
-	O
term	O
response	O
of	O
Swiss	B-Cell
3T3	I-Cell
cells	I-Cell
to	O
an	O
activator	O
of	O
protein	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
C	I-Gene_or_gene_product
also	O
appeared	O
to	O
be	O
independent	O
of	O
growth	O
state	O
,	O
since	O
the	O
50	O
%	O
effective	O
dose	O
for	O
PDBu	B-Simple_chemical
inhibition	O
of	O
EGF	B-Gene_or_gene_product
binding	O
to	O
its	O
receptor	O
was	O
approximately	O
7	O
nM	O
for	O
both	O
growth	O
conditions	O
.	O

In	O
contrast	O
,	O
the	O
response	O
of	O
cells	B-Cell
to	O
long	O
-	O
term	O
treatment	O
with	O
PDBu	B-Simple_chemical
was	O
significantly	O
different	O
depending	O
upon	O
the	O
initial	O
growth	O
state	O
of	O
the	O
cells	B-Cell
.	O

In	O
both	O
growth	O
states	O
,	O
PDBu	B-Simple_chemical
caused	O
loss	O
of	O
protein	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
C	I-Gene_or_gene_product
activity	O
,	O
which	O
reflected	O
a	O
loss	O
in	O
protein	O
mass	O
as	O
determined	O
by	O
immunoblotting	O
with	O
antiserum	B-Organism_substance
to	O
protein	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
C	I-Gene_or_gene_product
.	O

However	O
,	O
the	O
maximum	O
decrease	O
approached	O
100	O
%	O
in	O
stationary	O
cultures	O
versus	O
approximately	O
75	O
%	O
in	O
growing	O
cells	B-Cell
.	O

Protein	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
C	I-Gene_or_gene_product
levels	O
in	O
several	O
transformed	O
cell	B-Cell
lines	I-Cell
were	O
subject	O
to	O
down	O
modulation	O
in	O
a	O
similar	O
growth	O
state	O
-	O
dependent	O
manner	O
.	O

Further	O
,	O
the	O
inhibition	O
of	O
EGF	B-Gene_or_gene_product
binding	O
by	O
tumor	B-Cancer
promoters	O
following	O
long	O
-	O
term	O
treatment	O
of	O
Swiss	B-Cell
3T3	I-Cell
cells	I-Cell
with	O
PDBu	B-Simple_chemical
also	O
varied	O
with	O
growth	O
state	O
.	O

In	O
down	O
modulated	O
growing	O
cells	B-Cell
,	O
PDBu	B-Simple_chemical
caused	O
almost	O
complete	O
inhibition	O
of	O
EGF	B-Gene_or_gene_product
binding	O
,	O
whereas	O
in	O
down	O
modulated	O
stationary	O
cells	B-Cell
,	O
minimal	O
inhibition	O
of	O
EGF	B-Gene_or_gene_product
binding	O
by	O
PDBu	B-Simple_chemical
was	O
observed	O
.	O

These	O
results	O
suggest	O
that	O
prolonged	O
treatment	O
with	O
tumor	B-Cancer
promoters	O
alters	O
the	O
sensitivity	O
of	O
cells	B-Cell
to	O
activators	O
of	O
protein	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
C	I-Gene_or_gene_product
in	O
a	O
growth	O
state	O
-	O
dependent	O
manner	O
.	O

Suppression	O
and	O
re	O
-	O
expression	O
of	O
transformed	O
phenotype	O
in	O
hybrids	O
of	O
HA	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
transformed	O
rat	B-Cell
-	I-Cell
1	I-Cell
cells	I-Cell
and	O
early	O
-	O
passage	O
rat	B-Organism
embryonic	B-Cell
fibroblasts	I-Cell
.	O

Rat	B-Cell
-	I-Cell
1	I-Cell
cells	I-Cell
which	O
had	O
been	O
transformed	O
with	O
the	O
activated	O
Ha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
gene	O
from	O
human	B-Organism
EJ	B-Cell
bladder	I-Cell
carcinoma	I-Cell
cells	I-Cell
were	O
fused	O
with	O
diploid	O
embryonic	O
rat	B-Organism
fibroblasts	B-Cell
.	O

Four	O
selected	O
cell	B-Cell
hybrids	I-Cell
expressed	O
the	O
human	B-Organism
transforming	O
gene	O
product	O
p21	B-Gene_or_gene_product
at	O
levels	O
of	O
10	O
to	O
30	O
%	O
compared	O
to	O
100	O
%	O
in	O
the	O
transformed	O
parental	O
cells	B-Cell
.	O

The	O
hybrid	B-Cell
cells	I-Cell
,	O
however	O
,	O
exhibited	O
normal	O
morphology	O
,	O
anchorage	O
requirement	O
for	O
proliferation	O
,	O
and	O
largely	O
extended	O
latency	O
periods	O
of	O
tumorigenicity	O
in	O
newborn	O
rats	B-Organism
.	O

Tumorigenic	O
hybrid	B-Cell
derivatives	I-Cell
contained	O
lower	O
numbers	O
of	O
chromosomes	B-Cellular_component
than	O
the	O
tetraploid	O
parental	B-Cell
hybrids	I-Cell
.	O

DNA	B-Cellular_component
of	O
the	O
non	O
-	O
tumorigenic	O
cell	B-Cell
hybrids	I-Cell
transformed	O
Rat	B-Cell
-	I-Cell
1	I-Cell
cells	I-Cell
to	O
anchorage	O
-	O
independent	O
proliferation	O
as	O
expected	O
for	O
the	O
transforming	O
human	B-Organism
Ha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
gene	O
present	O
in	O
the	O
donor	O
DNA	B-Cellular_component
.	O

We	O
conclude	O
that	O
the	O
transforming	O
properties	O
of	O
the	O
activated	O
Ha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
gene	O
in	O
Rat	B-Cell
-	I-Cell
1	I-Cell
cells	I-Cell
can	O
be	O
suppressed	O
at	O
the	O
post	O
-	O
translational	O
level	O
by	O
the	O
presence	O
of	O
the	O
genome	B-Cellular_component
from	O
diploid	O
embryonic	O
rat	B-Organism
fibroblasts	B-Cell
but	O
additional	O
controls	O
of	O
expression	O
of	O
the	O
transforming	O
gene	O
are	O
likely	O
to	O
exist	O
.	O

Normal	B-Cell
cells	I-Cell
contain	O
suppressor	O
gene	O
(	O
s	O
)	O
which	O
safeguard	O
these	O
cells	B-Cell
against	O
transformation	O
by	O
the	O
product	O
of	O
the	O
transforming	O
Ha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
oncogene	O
.	O

Mast	B-Cell
cells	I-Cell
and	O
tumors	B-Cancer
.	O

The	O
specific	O
enhancement	O
of	O
tumor	B-Cancer
proliferation	O
in	O
vitro	O
.	O

Mast	B-Cell
cells	I-Cell
were	O
found	O
to	O
be	O
unique	O
among	O
the	O
peritoneal	B-Cell
leukocytes	I-Cell
by	O
virtue	O
of	O
their	O
capacity	O
to	O
enhance	O
profoundly	O
the	O
proliferation	O
of	O
a	O
variety	O
of	O
tumors	B-Cancer
in	O
vitro	O
.	O

This	O
phenomenon	O
occurs	O
at	O
mast	B-Cell
cell	I-Cell
/	O
tumor	B-Cancer
ratios	O
which	O
reflect	O
the	O
stoichiometry	O
of	O
host	B-Cell
cell	I-Cell
/	O
tumor	B-Cancer
relationships	O
in	O
vivo	O
.	O

The	O
growth	O
factor	O
was	O
found	O
to	O
reside	O
in	O
mast	B-Cell
cell	I-Cell
granules	B-Organism_substance
and	O
was	O
identified	O
as	O
heparin	B-Simple_chemical
by	O
sequential	O
purification	O
and	O
enzymatic	O
degradation	O
.	O

This	O
cellular	B-Cell
interaction	O
was	O
tumor	B-Cancer
-	O
specific	O
,	O
although	O
isolated	O
granules	B-Organism_substance
could	O
enhance	O
fibroblast	B-Cell
proliferation	O
.	O

The	O
findings	O
are	O
discussed	O
in	O
relation	O
to	O
previous	O
morphologic	O
studies	O
,	O
reports	O
of	O
in	O
vitro	O
mast	B-Cell
-	I-Cell
cell	I-Cell
-	O
mediated	O
tumor	B-Cancer
cytotoxicity	O
,	O
and	O
the	O
role	O
of	O
mast	B-Cell
cells	I-Cell
in	O
angiogenesis	O
and	O
connective	B-Tissue
tissue	I-Tissue
proliferation	O
.	O

Identification	O
of	O
the	O
putative	O
transforming	O
protein	O
of	O
the	O
human	B-Organism
T	I-Organism
-	I-Organism
cell	I-Organism
leukemia	I-Organism
viruses	I-Organism
HTLV	I-Organism
-	I-Organism
I	I-Organism
and	O
HTLV	B-Organism
-	I-Organism
II	I-Organism
.	O

The	O
human	B-Organism
T	I-Organism
-	I-Organism
cell	I-Organism
leukemia	I-Organism
viruses	I-Organism
HTLV	I-Organism
-	I-Organism
I	I-Organism
and	O
HTLV	B-Organism
-	I-Organism
II	I-Organism
are	O
unique	O
among	O
the	O
transforming	O
retroviruses	B-Organism
of	O
vertebrates	O
in	O
their	O
ability	O
to	O
transform	O
human	B-Organism
T	B-Cell
cells	I-Cell
in	O
vitro	O
and	O
in	O
their	O
close	O
association	O
with	O
human	B-Organism
malignancies	B-Cancer
(	O
T	B-Cancer
-	I-Cancer
cell	I-Cancer
lymphomas	I-Cancer
and	O
leukemia	B-Cancer
)	O
.	O

Their	O
genomes	O
are	O
relatively	O
simple	O
,	O
containing	O
the	O
genes	O
gag	B-Gene_or_gene_product
,	O
pol	B-Gene_or_gene_product
,	O
env	B-Gene_or_gene_product
,	O
and	O
a	O
3	B-Cellular_component
'	I-Cellular_component
region	I-Cellular_component
termed	O
"	O
X	B-Cellular_component
.	O
"	O
This	O
3	B-Cellular_component
'	I-Cellular_component
region	I-Cellular_component
may	O
be	O
responsible	O
for	O
the	O
transforming	O
potential	O
of	O
the	O
viruses	O
.	O

The	O
existence	O
of	O
proteins	O
encoded	O
by	O
the	O
3	B-Cellular_component
'	I-Cellular_component
region	I-Cellular_component
has	O
been	O
postulated	O
on	O
the	O
basis	O
of	O
multiple	O
open	O
reading	O
frames	O
.	O

In	O
the	O
present	O
study	O
this	O
region	O
is	O
shown	O
to	O
contain	O
a	O
gene	O
encoding	O
a	O
protein	O
of	O
40	O
kilodaltons	O
in	O
HTLV	B-Organism
-	I-Organism
I	I-Organism
and	O
37	O
kilodaltons	O
in	O
HTLV	B-Organism
-	I-Organism
II	I-Organism
.	O

It	O
is	O
proposed	O
that	O
these	O
proteins	O
be	O
called	O
,	O
respectively	O
,	O
p40xI	B-Gene_or_gene_product
and	O
p37xII	B-Gene_or_gene_product
.	O

Cells	B-Cell
transformed	O
by	O
Rous	B-Organism
sarcoma	I-Organism
virus	I-Organism
release	O
transforming	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factors	I-Gene_or_gene_product
.	O

Chicken	B-Organism
embryo	B-Cell
fibroblasts	I-Cell
and	O
hamster	B-Cell
BHK	I-Cell
cells	I-Cell
transformed	O
by	O
Rous	B-Organism
sarcoma	I-Organism
virus	I-Organism
(	O
RSV	B-Organism
)	O
release	O
in	O
their	O
culture	O
media	O
growth	O
factors	O
which	O
enhance	O
markedly	O
anchorage	O
-	O
independent	O
colony	O
formation	O
in	O
gelified	O
medium	O
,	O
at	O
the	O
restrictive	O
temperature	O
(	O
41	O
degrees	O
5	O
C	O
)	O
,	O
of	O
chicken	B-Organism
embryo	B-Cell
fibroblasts	I-Cell
(	O
CEF	B-Cell
)	O
infected	O
by	O
RSV	B-Organism
mutants	O
with	O
a	O
ts	O
mutation	O
of	O
the	O
src	B-Gene_or_gene_product
gene	O
.	O

This	O
action	O
is	O
not	O
observed	O
with	O
uninfected	O
CEF	B-Cell
,	O
and	O
,	O
therefore	O
,	O
appears	O
to	O
require	O
some	O
expression	O
of	O
the	O
viral	O
src	B-Gene_or_gene_product
gene	O
in	O
the	O
target	O
cells	B-Cell
.	O

The	O
enhancing	O
factors	O
are	O
proteins	O
related	O
to	O
the	O
family	O
of	O
the	O
transforming	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factors	I-Gene_or_gene_product
(	O
TGFs	B-Gene_or_gene_product
)	O
by	O
their	O
molecular	O
weight	O
(	O
about	O
20	O
kd	O
)	O
,	O
their	O
heat	O
and	O
acid	O
resistance	O
,	O
and	O
their	O
sensitivity	O
to	O
dithiothreitol	B-Simple_chemical
.	O

They	O
do	O
not	O
compete	O
with	O
125I	O
EGF	B-Gene_or_gene_product
for	O
binding	O
on	O
the	O
EGF	B-Gene_or_gene_product
receptors	I-Gene_or_gene_product
of	O
the	O
membrane	B-Cellular_component
of	O
A431	B-Cell
cells	I-Cell
.	O

As	O
chicken	B-Organism
embryo	B-Cell
fibroblasts	I-Cell
are	O
devoid	O
of	O
EGF	B-Gene_or_gene_product
receptors	I-Gene_or_gene_product
,	O
their	O
activity	O
is	O
not	O
potentiated	O
by	O
EGF	B-Gene_or_gene_product
.	O

The	O
genetics	O
of	O
transformation	O
by	O
SV	B-Organism
40	I-Organism
.	O

Experiments	O
using	O
the	O
tsA	B-Gene_or_gene_product
58	I-Gene_or_gene_product
allele	O
of	O
the	O
SV	B-Organism
40	I-Organism
-	O
A	B-Gene_or_gene_product
gene	O
have	O
demonstrated	O
that	O
the	O
SV	B-Organism
40	I-Organism
large	B-Gene_or_gene_product
T	I-Gene_or_gene_product
-	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
is	O
strictly	O
required	O
both	O
for	O
immortalization	O
and	O
induction	O
anchorage	O
independence	O
of	O
cells	B-Cell
.	O

It	O
has	O
been	O
suggested	O
that	O
the	O
immortalized	O
phenotype	O
is	O
mediated	O
by	O
an	O
extra	B-Immaterial_anatomical_entity
cellular	I-Immaterial_anatomical_entity
factor	B-Gene_or_gene_product
.	O

The	O
synthesis	O
and	O
/	O
or	O
the	O
excretion	O
of	O
this	O
factor	B-Gene_or_gene_product
in	O
a	O
medium	O
is	O
controlled	O
by	O
the	O
A	B-Gene_or_gene_product
gene	O
.	O

Molecular	O
events	O
leading	O
to	O
enhanced	O
glucose	B-Simple_chemical
transport	O
in	O
Rous	B-Organism
sarcoma	I-Organism
virus	I-Organism
-	O
transformed	O
cells	B-Cell
.	O

Transformation	O
by	O
Rous	B-Organism
sarcoma	I-Organism
virus	I-Organism
results	O
in	O
a	O
dramatic	O
increase	O
in	O
the	O
rate	O
at	O
which	O
the	O
transformed	O
cells	B-Cell
transport	O
glucose	B-Simple_chemical
across	O
the	O
cell	B-Cellular_component
membrane	I-Cellular_component
.	O

The	O
increased	O
transport	O
rate	O
is	O
a	O
consequence	O
of	O
an	O
increased	O
number	O
of	O
transporters	O
in	O
the	O
transformed	O
cells	B-Cell
.	O

Utilizing	O
antibody	O
raised	O
against	O
the	O
purified	O
human	B-Organism
erythrocyte	B-Cell
glucose	B-Simple_chemical
transporter	O
,	O
we	O
have	O
identified	O
the	O
glucose	B-Simple_chemical
transporter	O
as	O
a	O
membrane	B-Cellular_component
glycoprotein	O
with	O
a	O
monomer	O
Mr	O
of	O
approximately	O
41	O
,	O
000	O
.	O

The	O
increased	O
rate	O
of	O
glucose	B-Simple_chemical
transport	O
is	O
dependent	O
on	O
the	O
activity	O
of	O
pp60src	B-Gene_or_gene_product
,	O
the	O
transforming	O
protein	O
of	O
Rous	B-Organism
sarcoma	I-Organism
virus	I-Organism
.	O

This	O
protein	O
has	O
been	O
shown	O
to	O
be	O
a	O
protein	O
kinase	O
that	O
phosphorylates	O
on	O
tyrosine	O
residues	O
.	O

We	O
have	O
examined	O
the	O
tyrosine	B-Amino_acid
phosphorylation	O
of	O
a	O
major	O
cellular	B-Cell
protein	O
of	O
Mr	O
36	O
,	O
000	O
in	O
cells	B-Cell
infected	O
with	O
a	O
panel	O
of	O
partially	O
transforming	O
mutants	O
of	O
Rous	B-Organism
sarcoma	I-Organism
virus	I-Organism
.	O

One	O
of	O
these	O
mutants	O
(	O
CU2	B-Organism
)	O
increases	O
the	O
rate	O
of	O
glucose	B-Simple_chemical
transport	O
only	O
slightly	O
and	O
does	O
not	O
render	O
the	O
infected	O
cells	B-Cell
fully	O
anchorage	O
independent	O
or	O
tumorigenic	O
(	O
although	O
other	O
transformation	O
parameters	O
are	O
fully	O
induced	O
)	O
.	O

Cells	B-Cell
infected	O
with	O
this	O
mutant	O
display	O
a	O
36	O
,	O
000	O
-	O
dalton	O
protein	O
that	O
is	O
phosphorylated	O
to	O
a	O
considerably	O
lesser	O
extent	O
than	O
cells	B-Cell
infected	O
with	O
wild	B-Organism
-	I-Organism
type	I-Organism
virus	I-Organism
.	O

Analyses	O
of	O
this	O
sort	O
may	O
help	O
to	O
identify	O
the	O
cellular	B-Cell
targets	O
of	O
pp60src	B-Gene_or_gene_product
whose	O
phosphorylation	O
is	O
necessary	O
for	O
the	O
increased	O
glucose	B-Simple_chemical
transport	O
rate	O
.	O

DNA	B-Cellular_component
methylation	O
and	O
expression	O
of	O
HLA	B-Gene_or_gene_product
-	I-Gene_or_gene_product
DR	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
.	O

B	B-Cell
-	I-Cell
cell	I-Cell
lines	I-Cell
established	O
from	O
two	O
individuals	O
with	O
T	B-Cancer
-	I-Cancer
cell	I-Cancer
acute	I-Cancer
lymphocytic	I-Cancer
leukemia	I-Cancer
(	O
T	B-Cancer
-	I-Cancer
ALL	I-Cancer
)	O
express	O
HLA	B-Gene_or_gene_product
-	I-Gene_or_gene_product
DR	I-Gene_or_gene_product
antigens	I-Gene_or_gene_product
,	O
whereas	O
the	O
isogenic	O
T	B-Cell
-	I-Cell
cells	I-Cell
do	O
not	O
.	O

The	O
lack	O
of	O
expression	O
correlates	O
with	O
a	O
lack	O
of	O
detectable	O
HLA	B-Gene_or_gene_product
-	I-Gene_or_gene_product
DR	I-Gene_or_gene_product
mRNA	O
.	O

All	O
of	O
the	O
DR	B-Gene_or_gene_product
alpha	I-Gene_or_gene_product
DNA	B-Cellular_component
sequences	O
detected	O
by	O
a	O
cloned	O
DR	B-Gene_or_gene_product
alpha	I-Gene_or_gene_product
cDNA	O
probe	O
are	O
contained	O
in	O
a	O
BglII	B-Gene_or_gene_product
fragment	O
which	O
varies	O
slightly	O
in	O
size	O
(	O
4	O
.	O
0	O
to	O
4	O
.	O
8	O
kilobases	O
)	O
from	O
one	O
individual	O
to	O
another	O
.	O

In	O
DNA	B-Cellular_component
from	O
the	O
T	B-Cell
-	I-Cell
cells	I-Cell
not	O
expressing	O
DR	B-Gene_or_gene_product
alpha	I-Gene_or_gene_product
mRNA	O
,	O
all	O
of	O
the	O
potential	O
HpaII	B-Gene_or_gene_product
sites	O
within	O
the	O
BglII	B-Gene_or_gene_product
fragment	B-Cellular_component
appeared	O
to	O
be	O
methylated	O
.	O

In	O
contrast	O
,	O
at	O
least	O
some	O
of	O
these	O
sites	O
were	O
not	O
methylated	O
in	O
DNA	B-Cellular_component
from	O
the	O
B	B-Cell
-	I-Cell
cells	I-Cell
expressing	O
high	O
levels	O
of	O
DR	B-Gene_or_gene_product
alpha	I-Gene_or_gene_product
mRNA	O
.	O

Treatment	O
of	O
these	O
T	B-Cell
-	I-Cell
cells	I-Cell
with	O
5	B-Simple_chemical
-	I-Simple_chemical
azacytidine	I-Simple_chemical
resulted	O
in	O
the	O
induction	O
of	O
DR	B-Gene_or_gene_product
surface	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
expression	O
,	O
the	O
appearance	O
of	O
DR	B-Gene_or_gene_product
alpha	I-Gene_or_gene_product
mRNA	O
,	O
and	O
the	O
partial	O
demethylation	O
of	O
the	O
DR	B-Gene_or_gene_product
alpha	I-Gene_or_gene_product
DNA	B-Cellular_component
sequences	O
.	O

T	B-Cell
-	I-Cell
cell	I-Cell
lines	I-Cell
established	O
from	O
human	B-Organism
T	I-Organism
-	I-Organism
cell	I-Organism
leukemia	I-Organism
-	I-Organism
lymphoma	I-Organism
virus	I-Organism
associated	O
T	B-Pathological_formation
-	I-Pathological_formation
cell	I-Pathological_formation
neoplasias	I-Pathological_formation
,	O
in	O
contrast	O
to	O
the	O
T	B-Cell
-	I-Cell
cell	I-Cell
acute	I-Cell
lymphocytic	I-Cell
leukemia	I-Cell
cell	I-Cell
lines	I-Cell
,	O
expressed	O
both	O
DR	B-Gene_or_gene_product
antigens	I-Gene_or_gene_product
and	O
DR	B-Gene_or_gene_product
alpha	I-Gene_or_gene_product
mRNA	O
;	O
the	O
HpaII	B-Gene_or_gene_product
sites	O
within	O
the	O
BglII	B-Gene_or_gene_product
fragment	B-Cellular_component
of	O
DR	B-Gene_or_gene_product
alpha	I-Gene_or_gene_product
DNA	B-Cellular_component
of	O
these	O
human	B-Organism
T	I-Organism
-	I-Organism
cell	I-Organism
leukemia	I-Organism
-	I-Organism
lymphoma	I-Organism
virus	I-Organism
-	O
positive	O
T	B-Cell
-	I-Cell
cell	I-Cell
lines	I-Cell
were	O
in	O
all	O
cases	O
at	O
least	O
partially	O
unmethylated	O
.	O

Uncultured	O
peripheral	B-Cell
blood	I-Cell
T	I-Cell
-	I-Cell
cells	I-Cell
from	O
human	B-Organism
T	I-Organism
-	I-Organism
cell	I-Organism
leukemia	I-Organism
-	I-Organism
lymphoma	I-Organism
virus	I-Organism
-	O
infected	O
individuals	B-Organism
expressed	O
DR	B-Gene_or_gene_product
antigens	I-Gene_or_gene_product
at	O
a	O
low	O
level	O
,	O
and	O
the	O
DR	B-Gene_or_gene_product
alpha	I-Gene_or_gene_product
locus	B-Cellular_component
was	O
partially	O
unmethylated	O
.	O

After	O
48	O
h	O
in	O
culture	O
,	O
DR	B-Gene_or_gene_product
antigen	I-Gene_or_gene_product
expression	O
was	O
substantially	O
increased	O
,	O
but	O
no	O
significant	O
changes	O
were	O
observed	O
in	O
methylation	O
of	O
the	O
DR	B-Gene_or_gene_product
alpha	I-Gene_or_gene_product
locus	B-Cellular_component
or	O
in	O
the	O
amount	O
of	O
DR	B-Gene_or_gene_product
mRNA	O
which	O
was	O
present	O
.	O

This	O
suggests	O
that	O
expression	O
of	O
DR	B-Gene_or_gene_product
antigens	I-Gene_or_gene_product
also	O
can	O
be	O
modulated	O
post	O
-	O
transcriptionally	O
.	O

Hemostasis	O
and	O
mechanism	O
of	O
action	O
of	O
selective	O
antimetastatic	O
drugs	O
in	O
mice	B-Organism
bearing	O
Lewis	B-Cancer
lung	I-Cancer
carcinoma	I-Cancer
.	O

The	O
selective	O
antimetastatic	O
agents	O
p	B-Simple_chemical
-	I-Simple_chemical
(	I-Simple_chemical
3	I-Simple_chemical
,	I-Simple_chemical
3	I-Simple_chemical
-	I-Simple_chemical
dimethyl	I-Simple_chemical
-	I-Simple_chemical
1	I-Simple_chemical
-	I-Simple_chemical
triazeno	I-Simple_chemical
)	I-Simple_chemical
benzoic	I-Simple_chemical
acid	I-Simple_chemical
potassium	I-Simple_chemical
salt	I-Simple_chemical
(	O
DM	B-Simple_chemical
-	I-Simple_chemical
COOK	I-Simple_chemical
)	O
,	O
5	B-Simple_chemical
-	I-Simple_chemical
(	I-Simple_chemical
3	I-Simple_chemical
,	I-Simple_chemical
3	I-Simple_chemical
-	I-Simple_chemical
dimethyl	I-Simple_chemical
-	I-Simple_chemical
1	I-Simple_chemical
-	I-Simple_chemical
triazeno	I-Simple_chemical
)	I-Simple_chemical
imidazole	I-Simple_chemical
-	I-Simple_chemical
4	I-Simple_chemical
-	I-Simple_chemical
carboxamide	I-Simple_chemical
(	O
DTIC	B-Simple_chemical
)	O
and	O
(	B-Simple_chemical
+	I-Simple_chemical
/	I-Simple_chemical
-	I-Simple_chemical
)	I-Simple_chemical
1	I-Simple_chemical
,	I-Simple_chemical
2	I-Simple_chemical
-	I-Simple_chemical
di	I-Simple_chemical
(	I-Simple_chemical
3	I-Simple_chemical
,	I-Simple_chemical
5	I-Simple_chemical
-	I-Simple_chemical
dioxopiperazin	I-Simple_chemical
-	I-Simple_chemical
1	I-Simple_chemical
-	I-Simple_chemical
yl	I-Simple_chemical
)	I-Simple_chemical
propane	I-Simple_chemical
(	O
ICRF	B-Simple_chemical
-	I-Simple_chemical
159	I-Simple_chemical
)	O
have	O
been	O
shown	O
to	O
markedly	O
depress	O
the	O
formation	O
of	O
spontaneous	O
hematogenous	O
metastases	B-Cancer
in	O
mice	B-Organism
bearing	O
s	O
.	O
c	O
.	O

Lewis	B-Cancer
lung	I-Cancer
carcinoma	I-Cancer
,	O
with	O
a	O
mechanism	O
unrelated	O
to	O
cytotoxicity	O
for	O
tumor	B-Cell
cells	I-Cell
.	O

The	O
effects	O
on	O
hemostasis	O
of	O
DM	B-Simple_chemical
-	I-Simple_chemical
COOK	I-Simple_chemical
,	O
DTIC	B-Simple_chemical
and	O
ICRF	B-Simple_chemical
-	I-Simple_chemical
159	I-Simple_chemical
have	O
thus	O
been	O
examined	O
in	O
comparison	O
with	O
those	O
of	O
a	O
purely	O
cytotoxic	O
agent	O
,	O
cyclophosphamide	B-Simple_chemical
,	O
in	O
mice	B-Organism
bearing	O
i	O
.	O
m	O
.	O

Lewis	B-Cancer
lung	I-Cancer
carcinoma	I-Cancer
.	O

The	O
parameters	O
considered	O
are	O
the	O
number	O
of	O
platelets	B-Cell
and	O
their	O
aggregability	O
,	O
prothrombin	B-Gene_or_gene_product
and	O
partial	O
thromboplastin	B-Gene_or_gene_product
times	O
,	O
plasma	B-Organism_substance
fibrinogen	B-Gene_or_gene_product
concentration	O
and	O
tumor	B-Cell
cell	I-Cell
procoagulant	O
activity	O
.	O

Slight	O
variations	O
are	O
caused	O
by	O
drug	O
treatment	O
in	O
tumor	B-Cancer
-	O
bearing	O
mice	B-Organism
as	O
compared	O
with	O
untreated	O
tumor	B-Cancer
-	O
bearing	O
controls	B-Organism
;	O
the	O
pattern	O
of	O
effects	O
of	O
the	O
selective	O
antimetastatic	O
agents	O
does	O
not	O
differ	O
from	O
that	O
of	O
the	O
reference	O
cytotoxic	O
compound	O
used	O
,	O
cyclophosphamide	B-Simple_chemical
.	O

These	O
data	O
thus	O
indicate	O
that	O
the	O
effects	O
on	O
hemostasis	O
of	O
the	O
drugs	O
examined	O
can	O
contribute	O
only	O
marginally	O
to	O
their	O
antimetastatic	O
action	O
,	O
since	O
more	O
pronounced	O
effects	O
on	O
hemostasis	O
have	O
been	O
shown	O
to	O
be	O
required	O
to	O
significantly	O
affect	O
metastasis	O
formation	O
.	O

Metabolic	O
alterations	O
in	O
a	O
noncachectic	O
animal	O
tumor	B-Cancer
system	O
.	O

The	O
increased	O
energy	O
expended	O
by	O
the	O
host	O
to	O
synthesize	O
substrate	O
,	O
which	O
is	O
utilized	O
by	O
the	O
tumor	B-Cancer
,	O
is	O
a	O
potential	O
cause	O
of	O
cancer	B-Organism_substance
cachexia	I-Organism_substance
.	O

In	O
vivo	O
glucose	B-Simple_chemical
and	O
alanine	B-Amino_acid
kinetics	O
were	O
examined	O
by	O
tracer	O
methodology	O
in	O
a	O
sarcoma	B-Cancer
-	O
bearing	O
rat	B-Organism
model	O
.	O

The	O
effects	O
of	O
3	B-Simple_chemical
-	I-Simple_chemical
mercaptopicolinic	I-Simple_chemical
acid	I-Simple_chemical
,	O
a	O
potent	O
inhibitor	O
of	O
gluconeogenesis	O
,	O
was	O
also	O
examined	O
on	O
this	O
model	O
.	O

Both	O
tumor	B-Cancer
-	O
bearing	O
(	O
TB	O
)	O
and	O
nontumor	B-Cancer
bearing	O
(	O
NTB	O
)	O
animals	B-Organism
were	O
gaining	O
weight	O
prior	O
to	O
study	O
and	O
the	O
tumors	B-Cancer
were	O
relatively	O
small	O
.	O

The	O
TB	O
animals	B-Organism
had	O
significantly	O
lower	O
plasma	B-Organism_substance
glucose	B-Simple_chemical
and	O
higher	O
blood	B-Organism_substance
lactic	B-Simple_chemical
acid	I-Simple_chemical
levels	O
compared	O
with	O
NTB	O
animals	O
.	O

After	O
inhibition	O
of	O
gluconeogenesis	O
,	O
the	O
plasma	B-Organism_substance
glucose	B-Simple_chemical
decreased	O
and	O
the	O
blood	B-Organism_substance
lactate	B-Simple_chemical
increased	O
significantly	O
more	O
in	O
TB	O
than	O
NTB	O
animals	O
.	O

The	O
glucose	B-Simple_chemical
turnover	O
rate	O
was	O
significantly	O
greater	O
in	O
TB	O
compared	O
with	O
NTB	O
animals	O
,	O
as	O
was	O
the	O
rate	O
of	O
glucose	B-Simple_chemical
recycling	O
and	O
the	O
rate	O
of	O
gluconeogenesis	O
(	O
alanine	B-Amino_acid
leads	O
to	O
glucose	B-Simple_chemical
)	O
,	O
both	O
energy	O
demanding	O
processes	O
.	O

These	O
results	O
suggest	O
that	O
the	O
tumor	B-Cancer
-	O
bearing	O
animal	O
,	O
even	O
prior	O
to	O
significant	O
cachexia	B-Cancer
,	O
has	O
an	O
excess	O
demand	O
for	O
energy	O
,	O
the	O
provision	O
of	O
which	O
may	O
be	O
a	O
significant	O
factor	O
in	O
malignant	B-Cancer
cachexia	I-Cancer
.	O

The	O
inhibition	O
of	O
cultured	O
myoblast	B-Cell
differentiation	O
by	O
the	O
simian	B-Organism
virus	I-Organism
40	I-Organism
large	B-Gene_or_gene_product
T	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
occurs	O
after	O
myogenin	B-Gene_or_gene_product
expression	O
and	O
Rb	B-Gene_or_gene_product
up	O
-	O
regulation	O
and	O
is	O
not	O
exerted	O
by	O
transformation	O
-	O
competent	O
cytoplasmic	B-Organism_substance
mutants	O
.	O

We	O
have	O
investigated	O
the	O
mechanism	O
by	O
which	O
the	O
simian	B-Organism
virus	I-Organism
40	I-Organism
large	B-Gene_or_gene_product
T	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
(	O
SVLT	B-Gene_or_gene_product
)	O
interferes	O
with	O
the	O
differentiation	O
of	O
C2	B-Cell
myoblasts	I-Cell
.	O

SVLT	B-Gene_or_gene_product
mutants	O
,	O
defective	O
either	O
in	O
the	O
Rb	B-Gene_or_gene_product
binding	O
site	O
,	O
near	O
the	O
N	O
-	O
terminal	O
end	O
,	O
in	O
a	O
region	O
that	O
affects	O
binding	O
to	O
p53	B-Gene_or_gene_product
,	O
or	O
in	O
the	O
nuclear	O
transport	O
signal	O
,	O
were	O
also	O
employed	O
to	O
determine	O
whether	O
the	O
interference	O
was	O
especially	O
dependent	O
on	O
these	O
functional	O
domains	O
.	O

It	O
was	O
found	O
that	O
wild	O
-	O
type	O
(	O
wt	O
)	O
SVLT	B-Gene_or_gene_product
strongly	O
inhibited	O
the	O
terminal	O
differentiation	O
of	O
mouse	B-Organism
C2	B-Cell
myoblasts	I-Cell
,	O
but	O
this	O
arrest	O
occurred	O
only	O
after	O
the	O
synthesis	O
of	O
myogenin	B-Gene_or_gene_product
,	O
an	O
initial	O
step	O
in	O
biochemical	O
differentiation	O
.	O

Neither	O
the	O
synthesis	O
nor	O
some	O
basic	O
activities	O
of	O
MyoD	B-Gene_or_gene_product
appeared	O
to	O
be	O
affected	O
by	O
wt	O
SVLT	B-Gene_or_gene_product
.	O

In	O
these	O
transformants	B-Cell
,	O
mitogen	O
depletion	O
elicited	O
an	O
increase	O
in	O
the	O
Rb	B-Gene_or_gene_product
level	O
comparable	O
to	O
that	O
in	O
normal	O
C2	B-Cell
cells	I-Cell
;	O
wt	O
SVLT	B-Gene_or_gene_product
,	O
however	O
,	O
promoted	O
the	O
phosphorylation	O
of	O
a	O
large	O
part	O
of	O
the	O
induced	O
Rb	B-Gene_or_gene_product
.	O

Mutations	O
affecting	O
nuclear	B-Cellular_component
transport	O
were	O
far	O
more	O
critical	O
for	O
the	O
ability	O
to	O
interfere	O
with	O
myogenic	O
differentiation	O
than	O
were	O
those	O
affecting	O
the	O
transforming	O
potential	O
;	O
cytoplasmic	B-Organism_substance
SVLT	B-Gene_or_gene_product
expression	O
was	O
fully	O
compatible	O
with	O
the	O
terminal	O
differentiation	O
of	O
C2	B-Cell
cells	I-Cell
,	O
despite	O
enabling	O
them	O
to	O
grow	O
in	O
semisolid	O
medium	O
,	O
thus	O
showing	O
that	O
the	O
myogenesis	O
-	O
inhibiting	O
property	O
can	O
be	O
dissociated	O
from	O
transforming	O
competence	O
.	O

The	O
remaining	O
SVLT	B-Gene_or_gene_product
mutants	O
presented	O
different	O
degrees	O
of	O
ability	O
to	O
inhibit	O
differentiation	O
(	O
as	O
shown	O
by	O
the	O
expression	O
of	O
tissue	B-Tissue
-	O
specific	O
markers	O
in	O
transformants	B-Cell
)	O
.	O

The	O
inhibiting	O
mutants	O
,	O
including	O
the	O
Rb	B-Gene_or_gene_product
binding	O
site	O
mutant	O
,	O
were	O
able	O
to	O
promote	O
a	O
higher	O
state	O
of	O
Rb	B-Gene_or_gene_product
phosphorylation	O
than	O
that	O
observed	O
in	O
either	O
normal	O
cells	B-Cell
or	O
cytoplasmic	B-Organism_substance
-	O
SVLT	B-Gene_or_gene_product
transformants	B-Cell
.	O

A	O
transcript	O
from	O
the	O
long	B-Cellular_component
terminal	I-Cellular_component
repeats	I-Cellular_component
of	O
a	O
murine	B-Organism
retrovirus	I-Organism
associated	O
with	O
trans	O
activation	O
of	O
cellular	B-Cell
genes	O
.	O

Infection	O
of	O
human	B-Organism
or	O
murine	B-Organism
cells	B-Cell
with	O
murine	B-Organism
leukemia	I-Organism
viruses	I-Organism
rapidly	O
increases	O
the	O
expression	O
of	O
a	O
number	O
of	O
genes	O
that	O
belong	O
to	O
the	O
immunoglobulin	B-Gene_or_gene_product
superfamily	O
and	O
are	O
involved	O
in	O
T	B-Cell
-	I-Cell
lymphocyte	I-Cell
activation	O
,	O
including	O
the	O
class	B-Gene_or_gene_product
I	I-Gene_or_gene_product
major	I-Gene_or_gene_product
histocompatibility	I-Gene_or_gene_product
complex	I-Gene_or_gene_product
antigens	I-Gene_or_gene_product
.	O

We	O
have	O
reported	O
recently	O
that	O
the	O
long	B-Cellular_component
terminal	I-Cellular_component
repeat	I-Cellular_component
(	O
LTR	B-Cellular_component
)	O
of	O
Moloney	B-Organism
murine	I-Organism
leukemia	I-Organism
virus	I-Organism
encodes	O
a	O
trans	O
activator	O
which	O
induces	O
transcription	O
and	O
expression	O
of	O
class	B-Gene_or_gene_product
I	I-Gene_or_gene_product
major	I-Gene_or_gene_product
histocompatibility	I-Gene_or_gene_product
complex	I-Gene_or_gene_product
genes	O
and	O
certain	O
cytokine	B-Gene_or_gene_product
genes	O
.	O

The	O
portion	O
of	O
the	O
LTR	B-Cellular_component
responsible	O
for	O
trans	O
activation	O
was	O
mapped	O
by	O
deletions	O
to	O
lie	O
within	O
the	O
U3	O
region	O
.	O

We	O
demonstrate	O
here	O
that	O
a	O
transcript	O
is	O
initiated	O
within	O
the	O
U3	O
region	O
and	O
that	O
its	O
presence	O
correlates	O
with	O
the	O
trans	O
-	O
activating	O
activity	O
.	O

Analysis	O
of	O
the	O
LTR	B-Cellular_component
region	O
reveals	O
a	O
potential	O
internal	O
promoter	O
element	O
for	O
RNA	B-Gene_or_gene_product
polymerase	I-Gene_or_gene_product
III	I-Gene_or_gene_product
transcription	O
within	O
the	O
U3	O
region	O
.	O

Studies	O
with	O
polymerase	O
inhibitors	O
suggest	O
that	O
this	O
LTR	B-Cellular_component
transcript	O
,	O
designated	O
let	B-Gene_or_gene_product
(	O
LTR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
encoded	I-Gene_or_gene_product
trans	I-Gene_or_gene_product
activator	I-Gene_or_gene_product
)	O
,	O
is	O
a	O
product	O
of	O
RNA	B-Gene_or_gene_product
polymerase	I-Gene_or_gene_product
III	I-Gene_or_gene_product
.	O

The	O
mechanisms	O
whereby	O
RNA	B-Organism
leukemia	I-Organism
viruses	I-Organism
cause	O
lymphoid	B-Cancer
neoplasia	I-Cancer
after	O
a	O
long	O
latent	O
period	O
have	O
been	O
extensively	O
studied	O
but	O
are	O
only	O
partially	O
understood	O
.	O

The	O
region	O
of	O
the	O
LTR	B-Cellular_component
identified	O
here	O
as	O
being	O
important	O
in	O
trans	O
activation	O
has	O
recently	O
been	O
shown	O
to	O
be	O
a	O
critical	O
determinant	O
of	O
the	O
leukemogenicity	O
and	O
latency	O
of	O
Moloney	B-Organism
murine	I-Organism
leukemia	I-Organism
virus	I-Organism
.	O

These	O
findings	O
suggest	O
a	O
novel	O
mechanism	O
of	O
retrovirus	B-Organism
-	O
induced	O
activation	O
of	O
cellular	B-Cell
gene	O
expression	O
,	O
potentially	O
contributing	O
to	O
leukemogenesis	O
.	O

Anti	O
-	O
transforming	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	I-Gene_or_gene_product
TGF	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
antibodies	O
inhibit	O
breast	B-Cell
cancer	I-Cell
cell	I-Cell
tumorigenicity	O
and	O
increase	O
mouse	B-Organism
spleen	B-Cell
natural	I-Cell
killer	I-Cell
cell	I-Cell
activity	O
.	O

Implications	O
for	O
a	O
possible	O
role	O
of	O
tumor	B-Cell
cell	I-Cell
/	O
host	B-Cell
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
interactions	O
in	O
human	B-Organism
breast	B-Cancer
cancer	I-Cancer
progression	O
.	O

TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
effects	O
on	O
angiogenesis	O
,	O
stroma	B-Tissue
formation	O
,	O
and	O
immune	O
function	O
suggest	O
its	O
possible	O
involvement	O
in	O
tumor	B-Cancer
progression	O
.	O

This	O
hypothesis	O
was	O
tested	O
using	O
the	O
2G7	O
IgG2b	B-Gene_or_gene_product
,	O
which	O
neutralizes	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
-	B-Gene_or_gene_product
beta	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
and	O
-	B-Gene_or_gene_product
beta	I-Gene_or_gene_product
3	I-Gene_or_gene_product
,	O
and	O
the	O
MDA	B-Cell
-	I-Cell
231	I-Cell
human	B-Organism
breast	B-Cell
cancer	I-Cell
cell	I-Cell
line	I-Cell
.	O

Inoculation	O
of	O
these	O
cells	B-Cell
in	O
athymic	B-Organism
mice	I-Organism
decreases	O
mouse	B-Organism
spleen	B-Cell
natural	I-Cell
killer	I-Cell
(	I-Cell
NK	I-Cell
)	I-Cell
cell	I-Cell
activity	O
.	O

Intraperitoneal	B-Immaterial_anatomical_entity
injections	O
of	O
2G7	O
starting	O
1	O
d	O
after	O
intraperitoneal	B-Immaterial_anatomical_entity
inoculation	O
of	O
tumor	B-Cell
cells	I-Cell
suppressed	O
intraabdominal	B-Cancer
tumor	I-Cancer
and	O
lung	B-Organ
metastases	O
,	O
whereas	O
the	O
nonneutralizing	O
anti	O
-	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
12H5	O
IgG2a	O
had	O
no	O
effect	O
.	O

2G7	O
transiently	O
inhibited	O
growth	O
of	O
established	O
MDA	B-Cancer
-	I-Cancer
231	I-Cancer
subcutaneous	I-Cancer
tumors	I-Cancer
.	O

Histologically	O
,	O
both	O
2G7	O
-	O
treated	O
and	O
control	O
tumors	B-Cancer
were	O
identical	O
.	O

Intraperitoneal	B-Immaterial_anatomical_entity
administration	O
of	O
2G7	O
resulted	O
in	O
a	O
marked	O
increase	O
in	O
mouse	B-Organism
spleen	B-Cell
NK	I-Cell
cell	I-Cell
activity	O
.	O

2G7	O
did	O
not	O
inhibit	O
MDA	B-Cancer
-	I-Cancer
231	I-Cancer
primary	I-Cancer
tumor	I-Cancer
or	O
metastases	B-Cancer
formation	O
,	O
nor	O
did	O
it	O
stimulate	O
NK	B-Cell
cell	I-Cell
-	O
mediated	O
cytotoxicity	O
in	O
beige	B-Organism
NK	I-Organism
-	I-Organism
deficient	I-Organism
nude	I-Organism
mice	I-Organism
.	O

Finally	O
,	O
serum	B-Organism_substance
-	O
free	O
conditioned	O
medium	O
from	O
MDA	B-Cell
-	I-Cell
231	I-Cell
cells	I-Cell
inhibited	O
the	O
NK	B-Cell
cell	I-Cell
activity	O
of	O
human	B-Organism
blood	B-Cell
lymphocytes	I-Cell
.	O

This	O
inhibition	O
was	O
blocked	O
by	O
the	O
neutralizing	O
anti	O
-	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
2G7	O
antibody	O
but	O
not	O
by	O
a	O
nonspecific	O
IgG2	B-Gene_or_gene_product
.	O

These	O
data	O
support	O
a	O
possible	O
role	O
for	O
tumor	B-Cell
cell	I-Cell
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
in	O
the	O
progression	O
of	O
mammary	B-Cancer
carcinomas	I-Cancer
by	O
suppressing	O
host	O
immune	O
surveillance	O
.	O

Enhanced	O
DNA	B-Cellular_component
-	O
binding	O
activity	O
of	O
a	O
Stat3	B-Gene_or_gene_product
-	O
related	O
protein	O
in	O
cells	B-Cell
transformed	O
by	O
the	O
Src	B-Gene_or_gene_product
oncoprotein	O
.	O

Cytokines	B-Gene_or_gene_product
and	O
growth	B-Gene_or_gene_product
factors	I-Gene_or_gene_product
induce	O
tyrosine	B-Amino_acid
phosphorylation	O
of	O
signal	B-Gene_or_gene_product
transducers	I-Gene_or_gene_product
and	I-Gene_or_gene_product
activators	I-Gene_or_gene_product
of	I-Gene_or_gene_product
transcription	I-Gene_or_gene_product
(	O
STATs	B-Gene_or_gene_product
)	O
that	O
directly	O
activate	O
gene	O
expression	O
.	O

Cells	B-Cell
stably	O
transformed	O
by	O
the	O
Src	B-Gene_or_gene_product
oncogene	O
tyrosine	B-Amino_acid
kinase	O
were	O
examined	O
for	O
STAT	B-Gene_or_gene_product
protein	O
activation	O
.	O

Assays	O
of	O
electrophoretic	O
mobility	O
,	O
DNA	B-Cellular_component
-	O
binding	O
specificity	O
,	O
and	O
antigenicity	O
indicated	O
that	O
Stat3	B-Gene_or_gene_product
or	O
a	O
closely	O
related	O
STAT	B-Gene_or_gene_product
family	O
member	O
was	O
constitutively	O
activated	O
by	O
the	O
Src	B-Gene_or_gene_product
oncoprotein	O
.	O

Induction	O
of	O
this	O
DNA	B-Cellular_component
-	O
binding	O
activity	O
was	O
accompanied	O
by	O
tyrosine	B-Amino_acid
phosphorylation	O
of	O
Stat3	B-Gene_or_gene_product
and	O
correlated	O
with	O
Src	B-Gene_or_gene_product
transformation	O
.	O

These	O
findings	O
demonstrate	O
that	O
Src	B-Gene_or_gene_product
can	O
activate	O
STAT	B-Gene_or_gene_product
signaling	O
pathways	O
and	O
raise	O
the	O
possibility	O
that	O
Stat3	B-Gene_or_gene_product
contributes	O
to	O
oncogenesis	O
by	O
Src	B-Gene_or_gene_product
.	O

Bicistronic	O
retroviral	O
vector	O
reveals	O
capacity	O
of	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
erbA	I-Gene_or_gene_product
to	O
induce	O
erythroleukemia	B-Cancer
and	O
to	O
co	O
-	O
operate	O
with	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myb	I-Gene_or_gene_product
.	O

Previous	O
studies	O
have	O
shown	O
that	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
erbA	I-Gene_or_gene_product
and	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myb	I-Gene_or_gene_product
can	O
induce	O
the	O
proliferation	O
of	O
avian	O
erythroid	B-Cell
cells	I-Cell
in	O
culture	O
.	O

To	O
study	O
the	O
combined	O
effects	O
of	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
erbA	I-Gene_or_gene_product
and	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myb	I-Gene_or_gene_product
,	O
the	O
two	O
oncogenes	O
were	O
engineered	O
into	O
a	O
retrovirus	O
bicistronic	O
vector	O
with	O
an	O
internal	O
ribosomal	B-Cellular_component
entry	O
site	O
(	O
IRES	O
)	O
or	O
into	O
a	O
vector	O
with	O
a	O
splice	O
acceptor	O
(	O
SPL	O
)	O
.	O

This	O
allowed	O
coexpression	O
of	O
the	O
two	O
proteins	O
and	O
a	O
comparison	O
with	O
the	O
same	O
vector	O
containing	O
either	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
erbA	I-Gene_or_gene_product
or	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myb	I-Gene_or_gene_product
only	O
.	O

Both	O
the	O
erbA	B-Gene_or_gene_product
IRES	O
and	O
the	O
erbA	B-Gene_or_gene_product
/	O
myb	B-Gene_or_gene_product
IRES	O
virus	O
constructs	O
transformed	O
erythroid	B-Cell
cells	I-Cell
after	O
infection	O
of	O
bone	B-Multi-tissue_structure
marrow	I-Multi-tissue_structure
or	O
blastoderm	B-Cell
cultures	I-Cell
.	O

The	O
erbA	B-Gene_or_gene_product
/	O
myb	B-Gene_or_gene_product
IRES	O
virus	O
exhibited	O
a	O
5	O
-	O
10	O
-	O
fold	O
higher	O
transformed	O
colony	O
forming	O
efficiency	O
than	O
the	O
erbA	B-Gene_or_gene_product
IRES	O
virus	O
in	O
the	O
blastoderm	B-Cell
assay	O
.	O

Surprisingly	O
,	O
when	O
injected	O
into	O
chicken	B-Organism
embryos	B-Developing_anatomical_structure
in	O
the	O
presence	O
of	O
helper	O
virus	O
,	O
both	O
viruses	O
induced	O
an	O
erythroleukemia	B-Cancer
in	O
about	O
half	O
of	O
the	O
animals	O
.	O

In	O
contrast	O
,	O
no	O
leukemia	B-Cancer
was	O
observed	O
with	O
a	O
myb	B-Gene_or_gene_product
IRES	O
virus	O
,	O
with	O
spliced	O
vectors	O
containing	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
erbA	I-Gene_or_gene_product
alone	O
or	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
erbA	I-Gene_or_gene_product
in	O
combination	O
with	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myb	I-Gene_or_gene_product
,	O
nor	O
with	O
erbA	B-Gene_or_gene_product
IRES	O
and	O
erbA	B-Gene_or_gene_product
/	O
myb	B-Gene_or_gene_product
IRES	O
viruses	O
produced	O
in	O
the	O
absence	O
of	O
helper	O
virus	O
.	O

The	O
average	O
latency	O
of	O
leukemia	B-Cancer
induction	O
was	O
shorter	O
for	O
the	O
erbA	B-Gene_or_gene_product
/	O
myb	B-Gene_or_gene_product
IRES	O
virus	O
(	O
3	O
.	O
5	O
weeks	O
)	O
than	O
for	O
the	O
erbA	B-Gene_or_gene_product
IRES	O
virus	O
(	O
5	O
weeks	O
)	O
.	O

Nevertheless	O
,	O
for	O
both	O
viruses	O
the	O
leukemic	B-Cell
blasts	I-Cell
retained	O
full	O
factor	O
dependence	O
for	O
growth	O
.	O

These	O
results	O
show	O
that	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
erbA	I-Gene_or_gene_product
is	O
capable	O
of	O
inducing	O
an	O
erythroleukemia	B-Cancer
when	O
expressed	O
by	O
a	O
high	O
titer	O
bicistronic	O
retrovirus	O
under	O
conditions	O
of	O
virus	O
spreading	O
and	O
that	O
its	O
in	O
vitro	O
and	O
in	O
vivo	O
transforming	O
potential	O
can	O
be	O
enhanced	O
by	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myb	I-Gene_or_gene_product
.	O

VEGF165	B-Gene_or_gene_product
expressed	O
by	O
a	O
replication	O
-	O
deficient	O
recombinant	O
adenovirus	B-Organism
vector	O
induces	O
angiogenesis	O
in	O
vivo	O
.	O

To	O
evaluate	O
the	O
concept	O
that	O
localized	O
delivery	O
of	O
angiogenic	O
factors	O
via	O
virus	O
-	O
mediated	O
gene	O
transfer	O
may	O
be	O
useful	O
in	O
the	O
treatment	O
of	O
ischemic	O
disorders	O
,	O
the	O
replication	O
-	O
deficient	O
adenovirus	B-Organism
(	O
Ad	B-Organism
)	O
vector	O
AdCMV	O
.	O
VEGF165	B-Gene_or_gene_product
(	O
where	O
CMV	B-Organism
is	O
cytomegalovirus	B-Organism
and	O
VEGF	B-Gene_or_gene_product
is	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
)	O
containing	O
the	O
cDNA	B-Cellular_component
for	O
human	B-Organism
VEGF165	B-Gene_or_gene_product
,	O
a	O
secreted	O
endothelial	B-Cell
cell	I-Cell
-	O
specific	O
angiogenic	O
growth	O
factor	O
,	O
was	O
constructed	O
.	O

Human	B-Cell
umbilical	I-Cell
vein	I-Cell
endothelial	I-Cell
cells	I-Cell
(	O
HUVECs	B-Cell
)	O
and	O
rat	B-Cell
aorta	I-Cell
smooth	I-Cell
muscle	I-Cell
cells	I-Cell
(	O
RASMCs	B-Cell
)	O
infected	O
with	O
AdCMV	O
.	O
VEGF165	B-Gene_or_gene_product
(	O
5	O
and	O
20	O
plaque	O
-	O
forming	O
units	O
[	O
pfu	O
]	O
per	O
cell	B-Cell
)	O
demonstrated	O
VEGF	B-Gene_or_gene_product
mRNA	O
expression	O
and	O
protein	O
secretion	O
into	O
the	O
supernatant	O
.	O

Furthermore	O
,	O
the	O
conditioned	O
medium	O
from	O
these	O
cells	B-Cell
enhanced	O
vascular	B-Multi-tissue_structure
permeability	O
in	O
vivo	O
.	O

In	O
contrast	O
,	O
neither	O
VEGF	B-Gene_or_gene_product
mRNA	O
nor	O
secreted	O
protein	O
was	O
found	O
in	O
uninfected	O
HUVECs	B-Cell
or	O
RASMCs	B-Cell
or	O
in	O
cells	B-Cell
infected	O
with	O
the	O
control	O
vector	O
AdCMV	O
.	O
beta	B-Gene_or_gene_product
gal	I-Gene_or_gene_product
(	O
where	O
beta	B-Gene_or_gene_product
gal	I-Gene_or_gene_product
is	O
beta	B-Gene_or_gene_product
-	I-Gene_or_gene_product
galactosidase	I-Gene_or_gene_product
)	O
.	O

Assessment	O
of	O
starved	O
HUVECs	B-Cell
at	O
14	O
days	O
demonstrated	O
sixfold	O
more	O
cells	B-Cell
for	O
AdCMV	O
.	O
VEGF165	B-Gene_or_gene_product
-	O
infected	O
HUVECs	B-Cell
(	O
20	O
pfu	O
per	O
cell	B-Cell
)	O
than	O
for	O
either	O
infected	O
or	O
uninfected	O
control	O
cells	B-Cell
.	O

RASMC	B-Cell
proliferation	O
was	O
unaffected	O
by	O
infection	O
with	O
AdCMV	O
.	O
VEGF165	B-Gene_or_gene_product
.	O

When	O
plated	O
in	O
2	O
%	O
serum	B-Organism_substance
on	O
dishes	O
precoated	O
with	O
reconstituted	O
basement	B-Cellular_component
membrane	I-Cellular_component
(	O
Matrigel	O
)	O
,	O
HUVECs	B-Cell
infected	O
with	O
AdCMV	O
.	O
VEGF165	B-Gene_or_gene_product
(	O
20	O
pfu	O
per	O
cell	B-Cell
)	O
differentiated	O
into	O
capillary	B-Tissue
-	I-Tissue
like	I-Tissue
structures	I-Tissue
.	O

Under	O
similar	O
conditions	O
,	O
both	O
uninfected	O
HUVECs	B-Cell
and	O
HUVECs	B-Cell
infected	O
with	O
AdCMV	O
.	O
beta	B-Gene_or_gene_product
gal	I-Gene_or_gene_product
did	O
not	O
differentiate	O
.	O

To	O
evaluate	O
the	O
ability	O
of	O
AdCMV	O
.	O
VEGF165	B-Gene_or_gene_product
to	O
function	O
in	O
vivo	O
,	O
either	O
AdCMV	O
.	O

VEGF165	B-Gene_or_gene_product
or	O
AdCMV	O
.	O
beta	B-Gene_or_gene_product
gal	I-Gene_or_gene_product
(	O
2	O
x	O
10	O
(	O
10	O
)	O
pfu	O
)	O
was	O
resuspended	O
in	O
0	O
.	O
5	O
mL	O
Matrigel	O
and	O
injected	O
subcutaneously	O
into	O
mice	B-Organism
.	O

Immunohistochemical	O
staining	O
demonstrated	O
VEGF	B-Gene_or_gene_product
in	O
the	O
tissues	B-Tissue
surrounding	O
the	O
Matrigel	O
plugs	O
containing	O
AdCMV	O
.	O
VEGF165	B-Gene_or_gene_product
up	O
to	O
3	O
weeks	O
after	O
injection	O
,	O
whereas	O
no	O
VEGF	B-Gene_or_gene_product
was	O
found	O
in	O
the	O
control	O
plugs	O
with	O
AdCMV	O
.	O
beta	B-Gene_or_gene_product
gal	I-Gene_or_gene_product
.	O

Two	O
weeks	O
after	O
injection	O
,	O
there	O
was	O
histological	O
evidence	O
of	O
neovascularization	O
in	O
the	O
tissues	B-Tissue
surrounding	O
the	O
Matrigel	O
containing	O
AdCMV	O
.	O
VEGF165	B-Gene_or_gene_product
,	O
whereas	O
no	O
significant	O
angiogenesis	O
was	O
observed	O
in	O
response	O
to	O
AdCMV	O
.	O
beta	B-Gene_or_gene_product
gal	I-Gene_or_gene_product
.	O

Furthermore	O
,	O
the	O
Matrigel	O
plugs	O
with	O
AdCMV	O
.	O
VEGF165	B-Gene_or_gene_product
demonstrated	O
hemoglobin	B-Gene_or_gene_product
content	O
fourfold	O
higher	O
than	O
the	O
plugs	O
with	O
AdCMV	O
.	O
beta	B-Gene_or_gene_product
gal	I-Gene_or_gene_product
.	O

Together	O
,	O
these	O
in	O
vitro	O
and	O
in	O
vivo	O
studies	O
are	O
consistent	O
with	O
the	O
concept	O
that	O
Ad	B-Organism
vectors	O
may	O
provide	O
a	O
useful	O
strategy	O
for	O
efficient	O
local	O
delivery	O
of	O
VEGF165	B-Gene_or_gene_product
in	O
the	O
treatment	O
of	O
ischemic	O
diseases	O
.	O

Modulation	O
of	O
apoptosis	O
in	O
human	B-Organism
myeloid	B-Cell
leukemic	I-Cell
cells	I-Cell
by	O
GM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
.	O

Apoptosis	O
(	O
programmed	O
cell	B-Cell
death	O
)	O
regulates	O
cell	B-Cell
population	O
size	O
.	O

To	O
determine	O
the	O
mechanisms	O
whereby	O
hematopoietic	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factors	I-Gene_or_gene_product
(	O
HGFs	B-Gene_or_gene_product
)	O
modulate	O
apoptosis	O
in	O
human	B-Organism
myeloid	B-Cell
leukemic	I-Cell
cells	I-Cell
,	O
we	O
evaluated	O
the	O
roles	O
of	O
protein	O
and	O
mRNA	O
synthesis	O
for	O
altering	O
apoptosis	O
in	O
growth	O
factor	O
-	O
stimulated	O
vs	O
.	O
quiescent	O
leukemic	B-Cell
TF1	I-Cell
cells	I-Cell
.	O

Lysates	B-Organism_substance
of	O
cells	B-Cell
from	O
the	O
granulocyte	B-Gene_or_gene_product
-	I-Gene_or_gene_product
macrophage	I-Gene_or_gene_product
colony	I-Gene_or_gene_product
-	I-Gene_or_gene_product
stimulating	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
GM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
)	O
-	O
dependent	O
myeloid	B-Cell
leukemic	I-Cell
cell	I-Cell
line	I-Cell
TF1	I-Cell
were	O
separated	O
into	O
high	O
molecular	O
weight	O
(	O
HMW	O
)	O
pellets	O
of	O
intact	O
DNA	B-Cellular_component
and	O
supernatants	O
of	O
fragmented	O
low	O
MW	O
(	O
LMW	O
)	O
DNA	B-Cellular_component
,	O
and	O
the	O
DNA	B-Cellular_component
purified	O
from	O
these	O
fractions	O
was	O
quantified	O
.	O

In	O
the	O
absence	O
of	O
both	O
GM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
and	O
fetal	B-Organism_substance
bovine	I-Organism_substance
serum	I-Organism_substance
(	O
FBS	B-Organism_substance
)	O
,	O
70	O
%	O
of	O
the	O
DNA	B-Cellular_component
was	O
fragmented	O
after	O
3	O
days	O
in	O
culture	O
,	O
with	O
a	O
characteristic	O
apoptotic	O
ladder	O
-	O
like	O
pattern	O
on	O
agarose	B-Simple_chemical
gel	O
electrophoresis	O
,	O
whereas	O
this	O
proportion	O
had	O
initially	O
been	O
<	O
5	O
%	O
.	O

In	O
contrast	O
,	O
less	O
than	O
5	O
%	O
of	O
the	O
DNA	B-Cellular_component
was	O
fragmented	O
in	O
cells	B-Cell
incubated	O
with	O
GM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
plus	O
FBS	B-Organism_substance
or	O
GM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
alone	O
.	O

Delayed	O
addition	O
of	O
GM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
,	O
but	O
not	O
FBS	B-Organism_substance
,	O
permitted	O
partial	O
rescue	O
of	O
the	O
cells	B-Cell
,	O
inhibiting	O
increasing	O
rates	O
of	O
accumulation	O
of	O
fragmented	O
DNA	B-Cellular_component
.	O

When	O
the	O
macro	O
-	O
molecular	O
synthesis	O
inhibitor	O
cycloheximide	B-Simple_chemical
(	O
CHX	B-Simple_chemical
)	O
or	O
actinomycin	B-Simple_chemical
D	I-Simple_chemical
(	O
Act	B-Simple_chemical
D	I-Simple_chemical
)	O
was	O
present	O
for	O
26	O
hours	O
in	O
the	O
absence	O
of	O
GM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
and	O
FBS	B-Organism_substance
,	O
apoptosis	O
was	O
inhibited	O
.	O

In	O
contrast	O
,	O
in	O
the	O
presence	O
of	O
GM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
or	O
FBS	B-Organism_substance
,	O
apoptosis	O
was	O
enhanced	O
upon	O
addition	O
of	O
CHX	B-Simple_chemical
or	O
Act	B-Simple_chemical
D	I-Simple_chemical
.	O

The	O
latter	O
effect	O
persisted	O
even	O
with	O
the	O
late	O
addition	O
of	O
CHX	B-Simple_chemical
.	O

These	O
findings	O
indicate	O
that	O
disparate	O
mechanisms	O
of	O
enhancing	O
or	O
inhibiting	O
apoptosis	O
exist	O
in	O
myeloid	B-Cell
leukemic	I-Cell
cells	I-Cell
related	O
to	O
environmental	O
conditions	O
,	O
including	O
HGF	B-Gene_or_gene_product
-	O
regulated	O
cellular	B-Cell
synthesis	O
of	O
distinct	O
proteins	O
and	O
mRNA	O
.	O

Effects	O
of	O
doxycycline	B-Simple_chemical
on	O
in	O
vitro	O
growth	O
,	O
migration	O
,	O
and	O
gelatinase	O
activity	O
of	O
breast	B-Cell
carcinoma	I-Cell
cells	I-Cell
.	O

Metastatic	O
disease	O
is	O
one	O
of	O
the	O
major	O
causes	O
of	O
death	O
from	O
cancer	B-Cancer
in	O
human	B-Organism
beings	O
.	O

Several	O
enzyme	O
systems	O
have	O
been	O
implicated	O
in	O
the	O
metastatic	O
process	O
,	O
but	O
the	O
metalloproteinases	B-Gene_or_gene_product
(	O
MPs	B-Gene_or_gene_product
)	O
appear	O
to	O
be	O
the	O
major	O
group	O
involved	O
in	O
most	O
instances	O
of	O
neoplastic	O
invasion	O
.	O

Increased	O
MP	B-Gene_or_gene_product
activity	O
has	O
been	O
correlated	O
with	O
the	O
metastatic	O
potential	O
of	O
many	O
cancers	B-Cancer
,	O
including	O
breast	B-Cancer
cancer	I-Cancer
.	O

MPs	B-Gene_or_gene_product
also	O
play	O
a	O
role	O
in	O
tumor	B-Cancer
angiogenesis	O
.	O

Tetracyclines	B-Simple_chemical
are	O
antimicrobial	O
agents	O
that	O
can	O
suppress	O
MP	B-Gene_or_gene_product
activity	O
in	O
a	O
variety	O
of	O
tissues	B-Tissue
,	O
including	O
gingiva	B-Multi-tissue_structure
,	O
bone	B-Tissue
,	O
and	O
cartilage	B-Tissue
.	O

Several	O
reports	O
have	O
indicated	O
that	O
tetracyclines	B-Simple_chemical
can	O
suppress	O
tumor	B-Cancer
MPs	B-Gene_or_gene_product
as	O
well	O
.	O

A	O
synthetic	O
tetracycline	B-Simple_chemical
,	O
doxycycline	B-Simple_chemical
,	O
inhibits	O
migration	O
of	O
human	B-Organism
MDA	B-Cell
-	I-Cell
MB	I-Cell
-	I-Cell
435	I-Cell
breast	I-Cell
adenocarcinoma	I-Cell
cells	I-Cell
through	O
a	O
reconstituted	O
basement	B-Cellular_component
membrane	I-Cellular_component
(	O
Matrigel	O
)	O
,	O
an	O
assay	O
used	O
as	O
an	O
in	O
vitro	O
surrogate	O
for	O
the	O
in	O
vivo	O
process	O
of	O
tumor	B-Cancer
invasion	O
through	O
basement	B-Cellular_component
membranes	I-Cellular_component
.	O

Additionally	O
,	O
doxycycline	B-Simple_chemical
diminishes	O
the	O
proliferation	O
of	O
this	O
breast	B-Cell
cancer	I-Cell
cell	I-Cell
line	I-Cell
and	O
also	O
decreases	O
its	O
gelatinolytic	O
activity	O
,	O
as	O
determined	O
by	O
gel	O
zymography	O
.	O

Transforming	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
1	I-Gene_or_gene_product
promotes	O
spontaneous	O
transformation	O
of	O
cultured	O
rat	B-Organism
liver	B-Cell
epithelial	I-Cell
cells	I-Cell
.	O

The	O
neoplastic	O
transformation	O
of	O
cultured	O
rat	B-Organism
liver	B-Cell
epithelial	I-Cell
cells	I-Cell
by	O
various	O
means	O
has	O
consistently	O
been	O
associated	O
with	O
the	O
development	O
of	O
resistance	O
to	O
the	O
mito	O
-	O
inhibitory	O
effect	O
of	O
transforming	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
(	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
)	O
,	O
suggesting	O
that	O
such	O
phenotype	O
plays	O
a	O
mechanistic	O
role	O
during	O
the	O
transformation	O
of	O
these	O
cells	B-Cell
.	O

We	O
have	O
studied	O
the	O
induction	O
of	O
the	O
"	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
-	O
resistant	O
"	O
phenotype	O
in	O
a	O
clonal	O
strain	O
of	O
early	O
passage	O
WB	B-Cell
-	I-Cell
F344	I-Cell
normal	O
cultured	O
rat	B-Organism
liver	B-Cell
epithelial	I-Cell
cells	I-Cell
,	O
the	O
proliferation	O
of	O
which	O
was	O
markedly	O
inhibited	O
by	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
.	O

The	O
control	B-Cell
WB	I-Cell
cells	I-Cell
in	O
continuous	O
culture	O
slowly	O
developed	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
resistance	O
.	O

However	O
,	O
when	O
the	O
same	O
cells	B-Cell
were	O
exposed	O
to	O
step	O
-	O
wise	O
increases	O
of	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
concentration	O
in	O
their	O
culture	O
medium	O
,	O
the	O
development	O
of	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
resistance	O
was	O
accelerated	O
.	O

Cells	B-Cell
which	O
had	O
been	O
grown	O
in	O
medium	O
containing	O
1	O
ng	O
/	O
ml	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
developed	O
colony	O
-	O
forming	O
capacity	O
in	O
soft	O
agar	O
containing	O
epidermal	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
.	O

Cells	B-Cell
which	O
were	O
grown	O
in	O
media	O
containing	O
5	O
and	O
10	O
ng	O
/	O
ml	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
demonstrated	O
a	O
low	O
level	O
of	O
colony	O
-	O
forming	O
efficiency	O
in	O
soft	O
agar	O
medium	O
without	O
added	O
epidermal	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
and	O
tumorigenicity	O
in	O
isogeneic	O
rats	B-Organism
.	O

These	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
-	O
resistant	O
cells	B-Cell
also	O
exhibited	O
progressively	O
increasing	O
levels	O
of	O
expression	O
of	O
the	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
fos	I-Gene_or_gene_product
and	O
and	O
myc	B-Gene_or_gene_product
mRNA	O
,	O
and	O
increased	O
resistance	O
to	O
the	O
cytotoxicity	O
of	O
Adriamycin	B-Simple_chemical
and	O
melphalan	B-Simple_chemical
.	O

The	O
latter	O
phenomenon	O
was	O
accompanied	O
by	O
an	O
increase	O
in	O
the	O
mdr	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
mRNA	O
expression	O
,	O
cellular	B-Cell
glutathione	B-Simple_chemical
level	O
,	O
and	O
glutathione	B-Gene_or_gene_product
S	I-Gene_or_gene_product
-	I-Gene_or_gene_product
transferase	I-Gene_or_gene_product
activity	O
.	O

The	O
results	O
suggest	O
that	O
chronic	O
exposure	O
to	O
high	O
concentration	O
of	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
promotes	O
the	O
spontaneous	O
neoplastic	O
transformation	O
of	O
cultured	O
rat	B-Organism
liver	B-Cell
epithelial	I-Cell
cells	I-Cell
,	O
and	O
that	O
this	O
process	O
may	O
represent	O
one	O
of	O
the	O
mechanisms	O
of	O
cellular	B-Cell
adaptation	O
for	O
induction	O
of	O
the	O
multidrug	O
-	O
resistant	O
phenotype	O
during	O
the	O
carcinogenesis	O
of	O
epithelial	B-Cell
cells	I-Cell
.	O

Identification	O
of	O
latency	O
-	O
associated	O
transcripts	O
that	O
map	O
antisense	O
to	O
the	O
ICP4	B-Gene_or_gene_product
homolog	I-Gene_or_gene_product
gene	O
of	O
Marek	B-Organism
'	I-Organism
s	I-Organism
disease	I-Organism
virus	I-Organism
.	O

Two	O
small	O
RNAs	O
(	O
0	O
.	O
9	O
and	O
0	O
.	O
75	O
kb	O
)	O
,	O
named	O
Marek	B-Gene_or_gene_product
'	I-Gene_or_gene_product
s	I-Gene_or_gene_product
disease	I-Gene_or_gene_product
virus	I-Gene_or_gene_product
(	I-Gene_or_gene_product
MDV	I-Gene_or_gene_product
)	I-Gene_or_gene_product
small	I-Gene_or_gene_product
RNAs	I-Gene_or_gene_product
(	O
MSRs	B-Gene_or_gene_product
)	O
and	O
a	O
10	O
-	O
kb	O
RNA	O
,	O
all	O
of	O
which	O
map	O
antisense	O
to	O
the	O
MDV	B-Organism
ICP4	B-Gene_or_gene_product
homolog	I-Gene_or_gene_product
gene	O
,	O
have	O
been	O
readily	O
detected	O
in	O
MDCC	B-Cell
-	I-Cell
MSB1	I-Cell
MDV	B-Organism
-	O
transformed	O
T	B-Cell
-	I-Cell
lymphoblastoid	I-Cell
cells	I-Cell
.	O

These	O
RNAs	O
were	O
not	O
detectable	O
in	O
reticuloendotheliosis	B-Organism
virus	I-Organism
-	O
transformed	O
T	B-Cell
cells	I-Cell
.	O

When	O
MDV	B-Organism
was	O
reactivated	O
by	O
treatment	O
of	O
lymphoblastoid	B-Cell
cells	I-Cell
with	O
25	O
micrograms	O
of	O
iododeoxyuridine	B-Simple_chemical
per	O
ml	O
,	O
the	O
relative	O
levels	O
of	O
the	O
transcripts	O
decreased	O
.	O

These	O
RNAs	O
were	O
not	O
detected	O
by	O
Northern	O
(	O
RNA	O
)	O
hybridization	O
in	O
productively	O
infected	O
chicken	B-Organism
embryo	B-Cell
fibroblasts	I-Cell
48	O
h	O
postinfection	O
;	O
however	O
,	O
they	O
were	O
apparent	O
140	O
h	O
postinfection	O
.	O

By	O
using	O
Northern	O
hybridization	O
,	O
RNase	B-Gene_or_gene_product
protection	O
assays	O
,	O
and	O
primer	O
extension	O
analysis	O
,	O
the	O
MSRs	B-Gene_or_gene_product
were	O
determined	O
to	O
map	O
antisense	O
to	O
the	O
predicted	O
translational	O
start	O
site	O
of	O
the	O
ICP4	B-Gene_or_gene_product
homolog	I-Gene_or_gene_product
gene	O
.	O

The	O
conclusion	O
most	O
consistent	O
with	O
the	O
data	O
is	O
that	O
the	O
two	O
MSRs	B-Gene_or_gene_product
are	O
overlapping	O
,	O
spliced	O
RNAs	O
.	O

Both	O
small	O
RNAs	O
contain	O
a	O
latency	O
promoter	O
binding	O
factor	O
consensus	O
recognition	O
sequence	O
located	O
toward	O
their	O
5	O
'	O
ends	O
as	O
well	O
as	O
two	O
potential	O
ICP4	B-Gene_or_gene_product
recognition	O
consensus	O
sequences	O
,	O
one	O
in	O
each	O
orientation	O
.	O

The	O
region	O
contains	O
a	O
number	O
of	O
small	O
open	O
reading	O
frames	O
on	O
each	O
side	O
and	O
within	O
the	O
MSRs	B-Gene_or_gene_product
.	O

Although	O
the	O
exact	O
endpoints	O
are	O
unknown	O
,	O
the	O
large	O
10	O
-	O
kb	O
species	O
spans	O
the	O
entire	O
ICP4	B-Gene_or_gene_product
homolog	I-Gene_or_gene_product
region	O
.	O

We	O
believe	O
that	O
this	O
group	O
of	O
RNAs	O
,	O
which	O
map	O
antisense	O
to	O
the	O
ICP4	B-Gene_or_gene_product
homolog	I-Gene_or_gene_product
gene	O
,	O
are	O
latency	O
-	O
associated	O
transcripts	O
of	O
MDV	B-Organism
.	O

Increased	O
expression	O
of	O
specific	O
protein	B-Gene_or_gene_product
tyrosine	I-Gene_or_gene_product
phosphatases	I-Gene_or_gene_product
in	O
human	B-Organism
breast	B-Cell
epithelial	I-Cell
cells	I-Cell
neoplastically	O
transformed	O
by	O
the	O
neu	B-Gene_or_gene_product
oncogene	O
.	O

Protein	O
tyrosine	B-Amino_acid
phosphorylation	O
/	O
dephosphorylation	O
is	O
a	O
fundamental	O
mechanism	O
in	O
the	O
regulation	O
of	O
cell	B-Cell
proliferation	O
and	O
neoplastic	O
transformation	O
;	O
this	O
metabolic	O
process	O
is	O
modulated	O
by	O
the	O
opposing	O
activities	O
of	O
protein	B-Gene_or_gene_product
tyrosine	I-Gene_or_gene_product
kinases	I-Gene_or_gene_product
and	O
protein	B-Gene_or_gene_product
tyrosine	I-Gene_or_gene_product
phosphatases	I-Gene_or_gene_product
(	O
PTPases	B-Gene_or_gene_product
)	O
.	O

While	O
the	O
role	O
of	O
protein	B-Gene_or_gene_product
tyrosine	I-Gene_or_gene_product
kinases	I-Gene_or_gene_product
has	O
been	O
examined	O
extensively	O
in	O
human	B-Organism
breast	B-Organism_subdivision
tumorigenesis	O
,	O
the	O
role	O
of	O
PTPases	B-Gene_or_gene_product
in	O
this	O
process	O
is	O
virtually	O
unknown	O
.	O

To	O
address	O
this	O
issue	O
,	O
an	O
activated	O
neu	B-Gene_or_gene_product
oncogene	O
was	O
introduced	O
into	O
an	O
immortalized	O
nontumorigenic	O
human	B-Organism
breast	B-Cell
epithelial	I-Cell
cell	I-Cell
line	I-Cell
(	O
184B5	B-Cell
)	O
.	O

This	O
resulted	O
in	O
a	O
substantial	O
increase	O
in	O
P185neu	B-Gene_or_gene_product
expression	O
,	O
which	O
led	O
to	O
the	O
formation	O
of	O
progressively	O
growing	O
carcinomas	B-Cancer
after	O
such	O
cells	B-Cell
were	O
inoculated	O
into	O
athymic	O
nude	B-Organism
mice	I-Organism
.	O

Importantly	O
,	O
a	O
striking	O
increase	O
in	O
the	O
expression	O
of	O
specific	O
PTPases	B-Gene_or_gene_product
,	O
LAR	B-Gene_or_gene_product
and	O
PTP1B	B-Gene_or_gene_product
,	O
was	O
observed	O
in	O
3	O
independently	O
neu	B-Gene_or_gene_product
transformed	O
cell	B-Cell
lines	I-Cell
and	O
their	O
derived	O
tumors	B-Cancer
.	O

This	O
elevation	O
was	O
verified	O
at	O
both	O
the	O
mRNA	O
and	O
protein	O
levels	O
.	O

TC	B-Gene_or_gene_product
-	I-Gene_or_gene_product
PTP	I-Gene_or_gene_product
PTPase	I-Gene_or_gene_product
expression	O
was	O
only	O
slightly	O
increased	O
in	O
these	O
neu	B-Gene_or_gene_product
transformed	O
cells	B-Cell
,	O
and	O
no	O
expression	O
of	O
CD45	B-Gene_or_gene_product
PTPase	I-Gene_or_gene_product
was	O
observed	O
.	O

The	O
level	O
of	O
neu	B-Gene_or_gene_product
expression	O
,	O
as	O
well	O
as	O
the	O
differential	O
expression	O
between	O
P185neu	B-Gene_or_gene_product
and	O
LAR	B-Gene_or_gene_product
/	O
PTP1B	B-Gene_or_gene_product
,	O
directly	O
correlated	O
with	O
tumorigenicity	O
.	O

Furthermore	O
,	O
rat	B-Organism
mammary	B-Cancer
carcinomas	I-Cancer
with	O
elevated	O
neu	B-Gene_or_gene_product
expression	O
(	O
neu	B-Gene_or_gene_product
-	O
induced	O
)	O
also	O
had	O
sharply	O
elevated	O
LAR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
PTPase	I-Gene_or_gene_product
expression	O
when	O
compared	O
to	O
rat	B-Organism
mammary	B-Cancer
carcinomas	I-Cancer
with	O
little	O
or	O
no	O
neu	B-Gene_or_gene_product
expression	O
(	O
7	B-Simple_chemical
,	I-Simple_chemical
12	I-Simple_chemical
-	I-Simple_chemical
dimethylbenzanthracene	I-Simple_chemical
induced	O
)	O
;	O
the	O
level	O
of	O
expression	O
of	O
LAR	B-Gene_or_gene_product
PTPase	I-Gene_or_gene_product
was	O
directly	O
correlated	O
with	O
the	O
level	O
of	O
neu	B-Gene_or_gene_product
expression	O
.	O

Thus	O
,	O
our	O
results	O
provide	O
the	O
first	O
evidence	O
that	O
,	O
in	O
human	B-Organism
breast	B-Cell
carcinoma	I-Cell
cells	I-Cell
and	O
in	O
rat	B-Organism
mammary	B-Cancer
carcinomas	I-Cancer
that	O
have	O
an	O
induced	O
increase	O
in	O
neu	B-Gene_or_gene_product
expression	O
,	O
a	O
consistent	O
and	O
substantial	O
increase	O
in	O
the	O
expression	O
of	O
specific	O
PTPases	B-Gene_or_gene_product
occurs	O
.	O

The	O
relationship	O
between	O
P185neu	B-Gene_or_gene_product
-	O
protein	B-Gene_or_gene_product
tyrosine	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
expression	O
and	O
specific	O
PTPase	B-Gene_or_gene_product
expression	O
may	O
play	O
a	O
critical	O
role	O
in	O
human	B-Organism
breast	B-Organism_subdivision
tumorigenesis	O
.	O

In	O
vivo	O
effects	O
of	O
insulin	B-Gene_or_gene_product
on	O
tumor	B-Cancer
and	O
skeletal	B-Tissue
muscle	I-Tissue
glucose	B-Simple_chemical
metabolism	O
in	O
patients	B-Organism
with	O
lymphoma	B-Cancer
.	O

BACKGROUND	O
:	O
The	O
anabolic	O
properties	O
of	O
insulin	B-Gene_or_gene_product
have	O
been	O
suggested	O
for	O
use	O
to	O
reverse	O
malnutrition	O
associated	O
with	O
cancer	B-Cancer
.	O

The	O
host	O
and	O
tumor	B-Cancer
sensitivities	O
to	O
insulin	B-Gene_or_gene_product
are	O
critical	O
for	O
such	O
treatments	O
,	O
which	O
aim	O
to	O
improve	O
patient	B-Organism
nutrition	O
.	O

The	O
authors	O
studied	O
insulin	B-Gene_or_gene_product
effects	O
on	O
tumor	B-Cancer
and	O
skeletal	B-Tissue
muscle	I-Tissue
metabolism	O
with	O
2	B-Simple_chemical
-	I-Simple_chemical
[	I-Simple_chemical
18F	I-Simple_chemical
]	I-Simple_chemical
-	I-Simple_chemical
fluoro	I-Simple_chemical
-	I-Simple_chemical
2	I-Simple_chemical
-	I-Simple_chemical
deoxy	I-Simple_chemical
-	I-Simple_chemical
D	I-Simple_chemical
-	I-Simple_chemical
glucose	I-Simple_chemical
(	O
[	B-Simple_chemical
18F	I-Simple_chemical
]	I-Simple_chemical
FDG	I-Simple_chemical
)	O
and	O
positron	O
emission	O
tomography	O
(	O
PET	O
)	O
.	O

METHODS	O
:	O
Six	O
patients	B-Organism
with	O
lymphoma	B-Cancer
twice	O
underwent	O
[	B-Simple_chemical
18F	I-Simple_chemical
]	I-Simple_chemical
FDG	I-Simple_chemical
-	O
PET	O
imaging	O
:	O
once	O
after	O
fasting	O
overnight	O
and	O
once	O
during	O
euglycemic	O
hyperinsulinemic	O
clamp	O
.	O

The	O
dynamic	O
uptake	O
of	O
the	O
glucose	B-Simple_chemical
analogue	O
[	B-Simple_chemical
18F	I-Simple_chemical
]	I-Simple_chemical
FDG	I-Simple_chemical
was	O
measured	O
in	O
diseased	B-Multi-tissue_structure
nodes	I-Multi-tissue_structure
and	O
upper	B-Organism_subdivision
arm	I-Organism_subdivision
skeletal	B-Tissue
muscle	I-Tissue
in	O
both	O
metabolic	O
states	O
.	O

The	O
[	B-Simple_chemical
18F	I-Simple_chemical
]	I-Simple_chemical
FDG	I-Simple_chemical
uptake	O
in	O
muscle	B-Organ
and	O
the	O
whole	O
body	B-Organism_subdivision
glucose	B-Simple_chemical
use	O
during	O
euglycemic	O
hyperinsulinemic	O
clamp	O
were	O
compared	O
with	O
those	O
of	O
weight	O
-	O
matched	O
healthy	O
subjects	O
studied	O
under	O
similar	O
circumstances	O
.	O

RESULTS	O
:	O
In	O
lymphomatous	B-Tissue
tissue	I-Tissue
,	O
[	B-Simple_chemical
18F	I-Simple_chemical
]	I-Simple_chemical
FDG	I-Simple_chemical
uptake	O
rates	O
were	O
similar	O
in	O
overnight	O
fasting	O
and	O
euglycemic	O
hyperinsulinemic	O
clamp	O
(	O
38	O
+	O
/	O
-	O
10	O
versus	O
41	O
+	O
/	O
-	O
9	O
mumol	O
/	O
100	O
g	O
/	O
minute	O
,	O
not	O
significant	O
)	O
,	O
whereas	O
glucose	B-Simple_chemical
uptake	O
in	O
skeletal	B-Organ
muscle	I-Organ
was	O
increased	O
by	O
insulin	B-Gene_or_gene_product
(	O
1	O
.	O
7	O
+	O
/	O
-	O
0	O
.	O
2	O
versus	O
3	O
.	O
8	O
+	O
/	O
-	O
0	O
.	O
5	O
mumol	O
/	O
100	O
g	O
/	O
minute	O
,	O
P	O
=	O
0	O
.	O
012	O
)	O
.	O

Both	O
basal	O
(	O
2	O
.	O
3	O
+	O
/	O
-	O
0	O
.	O
2	O
mumol	O
/	O
100	O
g	O
/	O
minute	O
,	O
P	O
=	O
0	O
.	O
061	O
)	O
and	O
insulin	B-Gene_or_gene_product
-	O
stimulated	O
(	O
8	O
.	O
5	O
+	O
/	O
-	O
1	O
.	O
9	O
mumol	O
/	O
100	O
g	O
/	O
minute	O
,	O
P	O
=	O
0	O
.	O
055	O
)	O
skeletal	B-Multi-tissue_structure
arm	I-Multi-tissue_structure
muscle	I-Multi-tissue_structure
glucose	B-Simple_chemical
uptake	O
rates	O
were	O
higher	O
in	O
control	O
subjects	O
than	O
in	O
patients	B-Organism
.	O

Whole	O
body	B-Organism_subdivision
glucose	B-Simple_chemical
use	O
was	O
55	O
%	O
lower	O
in	O
patients	B-Organism
than	O
in	O
control	B-Organism
subjects	I-Organism
(	O
17	O
+	O
/	O
-	O
3	O
mumol	O
/	O
kg	O
/	O
minute	O
versus	O
38	O
+	O
/	O
-	O
3	O
mumol	O
/	O
kg	O
/	O
minute	O
,	O
P	O
=	O
0	O
.	O
002	O
)	O
,	O
consistent	O
with	O
insulin	B-Gene_or_gene_product
resistance	O
in	O
cancer	B-Cancer
.	O

CONCLUSIONS	O
:	O
We	O
found	O
that	O
insulin	B-Gene_or_gene_product
does	O
not	O
induce	O
major	O
changes	O
in	O
glucose	B-Simple_chemical
uptake	O
of	O
lymphomatous	B-Tissue
tissue	I-Tissue
.	O

Although	O
insulin	B-Gene_or_gene_product
sensitivity	O
of	O
skeletal	B-Organ
muscle	I-Organ
was	O
also	O
reduced	O
in	O
patients	B-Organism
with	O
lymphoma	B-Cancer
,	O
the	O
net	O
insulin	B-Gene_or_gene_product
effect	O
may	O
counteract	O
imbalance	O
between	O
glucose	B-Simple_chemical
uptake	O
of	O
tumor	B-Cancer
and	O
muscle	B-Organ
,	O
offering	O
a	O
potential	O
means	O
to	O
circumvent	O
at	O
least	O
some	O
metabolic	O
abnormalities	O
found	O
in	O
cancer	B-Cancer
.	O

Effect	O
of	O
1	B-Simple_chemical
-	I-Simple_chemical
alkylpyrrolidine	I-Simple_chemical
N	I-Simple_chemical
-	I-Simple_chemical
oxides	I-Simple_chemical
on	O
energy	O
metabolism	O
of	O
cancer	B-Cell
cells	I-Cell
.	O

The	O
main	O
purpose	O
of	O
the	O
present	O
investigation	O
was	O
to	O
study	O
the	O
effect	O
of	O
a	O
homologous	O
series	O
of	O
1	B-Simple_chemical
-	I-Simple_chemical
alkylpyrrolidine	I-Simple_chemical
N	I-Simple_chemical
-	I-Simple_chemical
oxides	I-Simple_chemical
on	O
ATP	B-Simple_chemical
-	O
producing	O
processes	O
in	O
Ehrlich	B-Cell
ascites	I-Cell
and	O
L1210	B-Cell
murine	I-Cell
leukemia	I-Cell
cells	I-Cell
.	O

The	O
effect	O
on	O
aerobic	O
glucose	B-Simple_chemical
consumption	O
,	O
lactic	B-Simple_chemical
acid	I-Simple_chemical
formation	O
,	O
content	O
of	O
total	O
(	O
T	O
-	O
SH	O
)	O
and	O
non	O
-	O
protein	O
thiol	O
groups	O
(	O
NP	O
-	O
SH	O
)	O
,	O
endogenous	O
respiration	O
and	O
the	O
level	O
of	O
ATP	B-Simple_chemical
in	O
tumor	B-Cell
cells	I-Cell
incubated	O
in	O
vitro	O
was	O
investigated	O
.	O

1	B-Simple_chemical
-	I-Simple_chemical
Tetradecylpyrrolidine	I-Simple_chemical
N	I-Simple_chemical
-	I-Simple_chemical
oxide	I-Simple_chemical
(	O
TPNO	B-Simple_chemical
)	O
,	O
one	O
of	O
the	O
most	O
active	O
compounds	O
,	O
immediately	O
after	O
addition	O
to	O
the	O
suspension	O
of	O
Ehrlich	B-Cell
cells	I-Cell
in	O
an	O
ice	O
bath	O
,	O
decreased	O
the	O
level	O
of	O
ATP	B-Simple_chemical
to	O
the	O
same	O
extent	O
over	O
the	O
whole	O
concentration	O
range	O
.	O

After	O
2	O
h	O
incubation	O
at	O
37	O
degrees	O
C	O
the	O
drop	O
in	O
the	O
ATP	B-Simple_chemical
level	O
was	O
lower	O
.	O

The	O
decrease	O
in	O
ATP	B-Simple_chemical
level	O
might	O
be	O
explained	O
through	O
the	O
interaction	O
of	O
the	O
amine	B-Simple_chemical
oxide	I-Simple_chemical
with	O
the	O
cell	B-Cellular_component
membrane	I-Cellular_component
integrity	O
.	O

Transformation	O
-	O
associated	O
cytokine	O
9E3	B-Gene_or_gene_product
/	O
CEF4	B-Gene_or_gene_product
is	O
chemotactic	O
for	O
chicken	B-Organism
peripheral	B-Cell
blood	I-Cell
mononuclear	I-Cell
cells	I-Cell
.	O

9E3	B-Gene_or_gene_product
/	O
CEF4	B-Gene_or_gene_product
,	O
which	O
is	O
released	O
from	O
transformed	O
chicken	B-Organism
embryo	B-Cell
fibroblasts	I-Cell
(	O
CEF	B-Cell
)	O
,	O
is	O
a	O
member	O
of	O
the	O
platelet	B-Gene_or_gene_product
factor	I-Gene_or_gene_product
4	I-Gene_or_gene_product
family	O
of	O
inflammatory	O
proteins	O
and	O
may	O
be	O
the	O
avian	O
homolog	O
of	O
interleukin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
.	O

Since	O
the	O
function	O
of	O
9E3	B-Gene_or_gene_product
/	O
CEF4	B-Gene_or_gene_product
is	O
unknown	O
,	O
we	O
examined	O
the	O
effect	O
of	O
the	O
protein	O
on	O
mitogenicity	O
and	O
chemotaxis	O
,	O
as	O
well	O
as	O
its	O
expression	O
,	O
in	O
fibroblasts	B-Cell
and	O
peripheral	B-Cell
blood	I-Cell
cells	I-Cell
.	O

9E3	B-Gene_or_gene_product
/	O
CEF4	B-Gene_or_gene_product
mRNA	O
was	O
expressed	O
in	O
chicken	B-Organism
peripheral	B-Organism_substance
blood	I-Organism_substance
monocytes	I-Organism_substance
,	O
and	O
its	O
expression	O
was	O
stimulated	O
by	O
incubation	O
of	O
the	O
monocytes	B-Cell
with	O
lipopolysaccharide	B-Simple_chemical
or	O
phorbol	B-Simple_chemical
myristic	I-Simple_chemical
acetate	I-Simple_chemical
.	O

Boyden	O
double	O
-	O
membrane	O
analysis	O
of	O
chemotaxis	O
showed	O
that	O
9E3	B-Gene_or_gene_product
/	O
CEF4	B-Gene_or_gene_product
was	O
chemotactic	O
for	O
chicken	B-Organism
peripheral	B-Cell
blood	I-Cell
mononuclear	I-Cell
cells	I-Cell
,	O
as	O
well	O
as	O
for	O
heterophils	B-Cell
.	O

Untransformed	O
CEF	B-Cell
and	O
CEF	B-Cell
transformed	O
with	O
Rous	B-Organism
sarcoma	I-Organism
virus	I-Organism
also	O
migrated	O
to	O
9E3	B-Gene_or_gene_product
/	O
CEF4	B-Gene_or_gene_product
protein	O
,	O
as	O
measured	O
by	O
Boyden	O
single	O
-	O
membrane	O
analysis	O
.	O

9E3	B-Gene_or_gene_product
/	O
CEF4	B-Gene_or_gene_product
was	O
slightly	O
mitogenic	O
for	O
CEF	B-Cell
,	O
causing	O
a	O
doubling	O
of	O
[	B-Simple_chemical
3H	I-Simple_chemical
]	I-Simple_chemical
thymidine	I-Simple_chemical
uptake	O
when	O
added	O
to	O
serum	B-Organism_substance
-	O
starved	O
CEF	B-Cell
.	O
9E3	B-Gene_or_gene_product
/	O
CEF4	B-Gene_or_gene_product
was	O
found	O
associated	O
not	O
only	O
with	O
the	O
cell	B-Cell
and	O
in	O
the	O
culture	O
medium	O
of	O
Rous	B-Organism
sarcoma	I-Organism
virus	I-Organism
-	O
transformed	O
CEF	B-Cell
but	O
also	O
with	O
the	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
.	O

The	O
in	O
vivo	O
role	O
of	O
9E3	B-Gene_or_gene_product
/	O
CEF4	B-Gene_or_gene_product
may	O
be	O
involved	O
with	O
chemotaxis	O
and	O
metastasis	O
,	O
rather	O
than	O
with	O
direct	O
stimulation	O
of	O
mitogenicity	O
.	O

Vitamins	B-Simple_chemical
regulate	O
gene	O
expression	O
and	O
induce	O
differentiation	O
and	O
growth	O
inhibition	O
in	O
cancer	B-Cell
cells	I-Cell
.	O

Their	O
relevance	O
in	O
cancer	B-Cancer
prevention	O
.	O

Although	O
several	O
hypotheses	O
for	O
human	B-Organism
carcinogenesis	O
have	O
been	O
proposed	O
,	O
the	O
specific	O
genetic	O
changes	O
that	O
cause	O
normal	O
cells	B-Cell
to	O
become	O
cancer	B-Cell
cells	I-Cell
have	O
not	O
been	O
identified	O
.	O

In	O
spite	O
of	O
uncertainties	O
regarding	O
the	O
mechanisms	O
of	O
carcinogenesis	O
,	O
several	O
vitamins	B-Simple_chemical
such	O
as	O
beta	B-Simple_chemical
-	I-Simple_chemical
carotene	I-Simple_chemical
and	O
vitamins	B-Simple_chemical
A	I-Simple_chemical
,	O
C	B-Simple_chemical
,	O
and	O
E	B-Simple_chemical
,	O
which	O
can	O
reduce	O
the	O
risk	O
of	O
cancer	B-Cancer
,	O
have	O
been	O
identified	O
,	O
using	O
animal	O
and	O
in	O
vitro	O
models	O
of	O
carcinogenesis	O
.	O

These	O
studies	O
have	O
led	O
to	O
a	O
hypothesis	O
that	O
the	O
supplemental	O
intake	O
of	O
these	O
vitamins	B-Simple_chemical
may	O
reduce	O
the	O
risk	O
of	O
cancer	B-Cancer
.	O

This	O
hypothesis	O
in	O
humans	B-Organism
can	O
be	O
tested	O
only	O
by	O
intervention	O
trials	O
that	O
are	O
in	O
progress	O
.	O

Prospective	O
and	O
retrospective	O
case	O
-	O
controlled	O
experimental	O
designs	O
are	O
not	O
suitable	O
for	O
testing	O
the	O
above	O
hypothesis	O
.	O

The	O
fact	O
that	O
some	O
vitamins	B-Simple_chemical
induce	O
cell	B-Cell
differentiation	O
and	O
/	O
or	O
growth	O
inhibition	O
in	O
tumor	B-Cell
cells	I-Cell
in	O
culture	O
suggests	O
that	O
the	O
use	O
of	O
these	O
vitamins	B-Simple_chemical
in	O
cancer	B-Cancer
prevention	O
has	O
a	O
cellular	B-Cell
basis	O
.	O

In	O
addition	O
to	O
having	O
a	O
direct	O
effect	O
on	O
tumor	B-Cell
cells	I-Cell
,	O
vitamins	B-Simple_chemical
such	O
as	O
alpha	B-Simple_chemical
-	I-Simple_chemical
tocopheryl	I-Simple_chemical
succinate	I-Simple_chemical
and	O
beta	B-Simple_chemical
-	I-Simple_chemical
carotene	I-Simple_chemical
enhance	O
the	O
effect	O
of	O
other	O
agents	O
that	O
induce	O
differentiation	O
in	O
tumor	B-Cell
cells	I-Cell
.	O

Some	O
vitamins	B-Simple_chemical
like	O
beta	B-Simple_chemical
-	I-Simple_chemical
carotene	I-Simple_chemical
,	O
retinoic	B-Simple_chemical
acid	I-Simple_chemical
,	O
alpha	B-Simple_chemical
-	I-Simple_chemical
tocopheryl	I-Simple_chemical
succinate	I-Simple_chemical
,	O
and	O
vitamin	B-Simple_chemical
D	I-Simple_chemical
also	O
regulate	O
the	O
expressions	O
of	O
certain	O
oncogenes	O
and	O
cellular	B-Cell
genes	O
.	O

These	O
are	O
exciting	O
new	O
functions	O
of	O
vitamins	B-Simple_chemical
that	O
nobody	O
could	O
have	O
predicted	O
only	O
a	O
few	O
years	O
ago	O
.	O

Increased	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
and	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H2	I-Gene_or_gene_product
messenger	O
RNA	O
expression	O
and	O
absence	O
of	O
mutations	O
in	O
colon	B-Cancer
carcinomas	I-Cancer
of	O
low	O
and	O
high	O
metastatic	O
potential	O
.	O

BACKGROUND	O
:	O
The	O
murine	B-Organism
nm23	B-Gene_or_gene_product
gene	O
suppresses	O
the	O
metastatic	O
behavior	O
of	O
malignant	B-Cell
rodent	I-Cell
tumor	I-Cell
lines	I-Cell
,	O
and	O
reduced	O
nm23	B-Gene_or_gene_product
expression	O
correlates	O
with	O
increased	O
likelihood	O
of	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastases	O
in	O
human	B-Organism
breast	B-Cancer
cancers	I-Cancer
.	O

More	O
recent	O
data	O
have	O
demonstrated	O
the	O
existence	O
of	O
two	O
human	B-Organism
nm23	B-Gene_or_gene_product
gene	O
homologues	O
,	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
and	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H2	I-Gene_or_gene_product
,	O
and	O
have	O
shown	O
that	O
deletion	O
of	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
alleles	O
occurs	O
in	O
some	O
colon	B-Cancer
carcinomas	I-Cancer
associated	O
with	O
poor	O
prognosis	O
.	O

These	O
findings	O
suggest	O
that	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
encodes	O
for	O
suppression	O
of	O
colon	B-Cancer
carcinoma	I-Cancer
metastasis	O
.	O

In	O
contrast	O
,	O
we	O
have	O
previously	O
reported	O
that	O
total	O
nm23	B-Gene_or_gene_product
messenger	O
RNA	O
(	O
mRNA	O
)	O
expression	O
is	O
increased	O
to	O
similar	O
levels	O
in	O
colon	B-Cancer
tumors	I-Cancer
of	O
both	O
high	O
and	O
low	O
metastatic	O
potential	O
.	O

PURPOSE	O
:	O
This	O
study	O
was	O
designed	O
to	O
reconcile	O
our	O
previous	O
findings	O
with	O
the	O
recent	O
report	O
of	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
allelic	O
deletion	O
in	O
human	B-Organism
colon	B-Cancer
cancers	I-Cancer
associated	O
with	O
poor	O
prognosis	O
.	O

Our	O
purpose	O
was	O
to	O
examine	O
human	B-Organism
colon	B-Cancer
cancers	I-Cancer
for	O
inactivation	O
of	O
two	O
candidate	O
metastasis	O
suppressor	O
genes	O
,	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
and	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H2	I-Gene_or_gene_product
,	O
either	O
by	O
mutation	O
or	O
by	O
loss	O
of	O
gene	O
transcription	O
.	O

METHODS	O
:	O
We	O
used	O
ribonuclease	B-Gene_or_gene_product
protection	O
assays	O
to	O
analyze	O
human	B-Organism
colon	B-Cancer
tumors	I-Cancer
for	O
the	O
level	O
of	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
(	O
43	O
samples	B-Cancer
)	O
and	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H2	I-Gene_or_gene_product
(	O
41	O
samples	B-Cancer
)	O
transcript	O
(	O
mRNA	O
)	O
expression	O
and	O
the	O
presence	O
of	O
mutations	O
that	O
could	O
inactivate	O
potential	O
suppressor	O
function	O
.	O

RESULTS	O
:	O
We	O
detected	O
only	O
wild	O
-	O
type	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
and	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H2	I-Gene_or_gene_product
mRNA	O
.	O

Expression	O
of	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
mRNA	O
increased	O
in	O
33	O
of	O
41	O
colon	B-Cancer
tumors	I-Cancer
,	O
and	O
expression	O
of	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H2	I-Gene_or_gene_product
mRNA	O
was	O
elevated	O
in	O
28	O
of	O
41	O
colon	B-Cancer
tumors	I-Cancer
relative	O
to	O
that	O
in	O
matched	O
normal	O
mucosa	B-Multi-tissue_structure
.	O

Increases	O
in	O
these	O
mRNA	O
levels	O
were	O
similar	O
in	O
tumors	B-Cancer
of	O
both	O
low	O
and	O
high	O
metastatic	O
potential	O
.	O

CONCLUSIONS	O
:	O
These	O
results	O
suggest	O
that	O
,	O
despite	O
correlation	O
of	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
allelic	O
deletions	O
with	O
colon	B-Cancer
cancers	I-Cancer
associated	O
with	O
poor	O
prognosis	O
,	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
and	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H2	I-Gene_or_gene_product
alleles	O
do	O
not	O
directly	O
mediate	O
metastasis	O
suppression	O
in	O
colon	B-Cancer
carcinoma	I-Cancer
.	O

Our	O
results	O
leave	O
unexplained	O
the	O
observation	O
that	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
allelic	O
deletion	O
correlates	O
with	O
metastatic	O
potential	O
of	O
colon	B-Cancer
carcinomas	I-Cancer
.	O

IMPLICATIONS	O
:	O
These	O
findings	O
also	O
contrast	O
with	O
the	O
demonstration	O
of	O
nm23	B-Gene_or_gene_product
metastasis	O
suppressor	O
activity	O
in	O
murine	B-Organism
melanoma	B-Cancer
and	O
with	O
the	O
correlation	O
of	O
loss	O
of	O
nm23	B-Gene_or_gene_product
expression	O
in	O
breast	B-Cancer
cancer	I-Cancer
with	O
poor	O
prognosis	O
.	O

It	O
may	O
be	O
that	O
metastasis	O
suppression	O
by	O
the	O
nm23	B-Gene_or_gene_product
gene	O
is	O
a	O
tissue	B-Tissue
-	O
specific	O
phenomenon	O
.	O

E	B-Gene_or_gene_product
mu	I-Gene_or_gene_product
-	I-Gene_or_gene_product
bcl	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
transgene	O
facilitates	O
spontaneous	O
transformation	O
of	O
early	O
pre	B-Cell
-	I-Cell
B	I-Cell
and	O
immunoglobulin	B-Gene_or_gene_product
-	O
secreting	O
cells	B-Cell
but	O
not	O
T	B-Cell
cells	I-Cell
.	O

To	O
assess	O
the	O
lymphoid	B-Cell
tumorigenic	O
potential	O
of	O
bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
mice	B-Organism
of	O
five	O
independent	O
strains	O
expressing	O
a	O
bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
transgene	O
in	O
B	B-Cell
and	O
/	O
or	O
T	B-Cell
cells	I-Cell
were	O
monitored	O
for	O
disease	O
up	O
to	O
12	O
months	O
of	O
age	O
.	O

Lymphoma	B-Cancer
prevalence	O
was	O
minimal	O
in	O
the	O
T	B-Cell
lineage	I-Cell
but	O
significant	O
,	O
although	O
low	O
(	O
3	O
-	O
15	O
%	O
)	O
,	O
in	O
the	O
B	B-Cell
lineage	I-Cell
.	O

The	O
principal	O
types	O
of	O
tumors	B-Cancer
were	O
plasmacytomas	B-Cancer
secreting	O
immunoglobulin	B-Gene_or_gene_product
and	O
novel	O
lymphomas	B-Cancer
that	O
expressed	O
markers	O
such	O
as	O
Sca	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
CD4	B-Gene_or_gene_product
,	O
Thy	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
CD34	B-Gene_or_gene_product
and	O
CD45	B-Gene_or_gene_product
(	O
B220	B-Gene_or_gene_product
)	O
,	O
consistent	O
with	O
an	O
origin	O
very	O
early	O
in	O
B	B-Cell
-	I-Cell
lymphoid	I-Cell
development	O
.	O

Rearrangement	O
of	O
the	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
gene	O
was	O
common	O
in	O
the	O
plasmacytomas	B-Cancer
,	O
implying	O
a	O
synergistic	O
role	O
for	O
myc	B-Gene_or_gene_product
and	O
bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
in	O
their	O
etiology	O
,	O
but	O
was	O
not	O
detected	O
in	O
the	O
lymphomas	B-Cancer
.	O

Tumorigenic	O
activity	O
of	O
rho	B-Gene_or_gene_product
genes	O
from	O
Aplysia	B-Organism
californica	I-Organism
.	O

rho	B-Gene_or_gene_product
genes	O
have	O
been	O
found	O
in	O
both	O
lower	O
and	O
higher	O
eucaryotes	O
.	O

They	O
code	O
for	O
proteins	O
of	O
21	O
kDa	O
,	O
highly	O
conserved	O
in	O
evolution	O
,	O
which	O
belong	O
to	O
the	O
superfamily	O
of	O
ras	B-Gene_or_gene_product
GTPases	I-Gene_or_gene_product
.	O

Among	O
the	O
members	O
of	O
this	O
superfamily	O
there	O
are	O
proteins	O
with	O
a	O
regulatory	O
function	O
,	O
such	O
as	O
ras	B-Gene_or_gene_product
,	O
and	O
proteins	O
involved	O
in	O
vesicular	B-Multi-tissue_structure
trafficking	O
,	O
such	O
as	O
the	O
family	O
of	O
rab	B-Gene_or_gene_product
proteins	O
.	O

We	O
have	O
investigated	O
the	O
putative	O
role	O
of	O
rho	B-Gene_or_gene_product
proteins	O
from	O
Aplysia	B-Organism
californica	I-Organism
as	O
transforming	O
GTPases	B-Gene_or_gene_product
utilizing	O
the	O
wild	O
-	O
type	O
and	O
a	O
Val	B-Organism
-	I-Organism
14	I-Organism
mutant	I-Organism
,	O
equivalent	O
to	O
the	O
oncogenic	O
Val	O
-	O
12	O
mutation	O
of	O
ras	B-Gene_or_gene_product
genes	O
found	O
in	O
animal	O
and	O
human	B-Organism
tumors	B-Cancer
.	O

Over	O
-	O
expression	O
of	O
either	O
rho	B-Gene_or_gene_product
gene	O
was	O
sufficient	O
to	O
confer	O
anchorage	O
-	O
and	O
serum	B-Organism_substance
-	O
independent	O
growth	O
.	O

Moreover	O
,	O
when	O
introduced	O
into	O
nude	B-Organism
mice	I-Organism
,	O
selected	O
clones	O
generated	O
from	O
either	O
gene	O
were	O
able	O
to	O
induce	O
tumors	B-Cancer
,	O
although	O
those	O
carrying	O
the	O
mutated	O
version	O
were	O
more	O
efficient	O
.	O

Pathological	O
analysis	O
indicated	O
that	O
generated	O
tumors	B-Cancer
corresponded	O
to	O
well	O
-	O
differentiated	O
fibrosarcomas	B-Cancer
with	O
distinct	O
and	O
intersecting	O
bundles	B-Cell
and	O
spindle	B-Cell
cells	I-Cell
.	O

By	O
contrast	O
,	O
ras	B-Gene_or_gene_product
-	O
induced	O
tumors	B-Cancer
were	O
poorly	O
differentiated	O
fibrosarcomas	B-Cancer
.	O

Thus	O
,	O
our	O
results	O
indicate	O
that	O
under	O
appropriate	O
conditions	O
rho	B-Gene_or_gene_product
genes	O
function	O
as	O
oncogenes	O
and	O
may	O
have	O
a	O
role	O
in	O
the	O
regulation	O
of	O
proliferation	O
in	O
fibroblast	B-Cell
cells	I-Cell
.	O

Pancreatic	B-Organ
somatostatin	B-Gene_or_gene_product
-	O
secreting	O
gangliocytic	B-Cancer
paraganglioma	I-Cancer
with	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastases	O
.	O

Gangliocytic	B-Cancer
paraganglioma	I-Cancer
(	O
GPG	B-Cancer
)	O
with	O
local	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
was	O
found	O
in	O
the	O
pancreas	B-Organ
of	O
a	O
74	O
-	O
yr	O
-	O
old	O
female	B-Organism
who	O
presented	O
with	O
diarrhea	O
,	O
steatorrhea	O
,	O
vomiting	O
,	O
nausea	O
,	O
and	O
abdominal	B-Organism_subdivision
pain	O
.	O

A	O
Whipple	O
procedure	O
led	O
to	O
a	O
complete	O
resolution	O
of	O
these	O
symptoms	O
and	O
a	O
return	O
of	O
an	O
elevated	O
stomatostatin	B-Gene_or_gene_product
level	O
to	O
normal	O
.	O

This	O
is	O
the	O
first	O
description	O
of	O
GPG	B-Cancer
in	O
this	O
location	O
and	O
the	O
first	O
endocrinologically	O
active	O
GPG	B-Cancer
.	O

Stimulation	O
of	O
endothelial	B-Cell
cell	I-Cell
migration	O
by	O
vascular	B-Gene_or_gene_product
permeability	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
/	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
through	O
cooperative	O
mechanisms	O
involving	O
the	O
alphavbeta3	B-Gene_or_gene_product
integrin	I-Gene_or_gene_product
,	O
osteopontin	B-Gene_or_gene_product
,	O
and	O
thrombin	B-Gene_or_gene_product
.	O

We	O
have	O
identified	O
several	O
mechanisms	O
by	O
which	O
the	O
angiogenic	O
cytokine	O
vascular	B-Gene_or_gene_product
permeability	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
/	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VPF	B-Gene_or_gene_product
/	O
VEGF	B-Gene_or_gene_product
)	O
likely	O
regulates	O
endothelial	B-Cell
cells	I-Cell
(	O
EC	B-Cell
)	O
migration	O
.	O

VPF	B-Gene_or_gene_product
/	O
VEGF	B-Gene_or_gene_product
induced	O
dermal	B-Cell
microvascular	I-Cell
EC	I-Cell
expression	O
of	O
mRNAs	O
encoding	O
the	O
alphav	B-Gene_or_gene_product
and	O
beta3	B-Gene_or_gene_product
integrin	I-Gene_or_gene_product
subunits	I-Gene_or_gene_product
resulting	O
in	O
increased	O
levels	O
of	O
the	O
alphavbeta3	B-Gene_or_gene_product
heterodimer	O
at	O
the	O
cell	B-Cellular_component
surface	I-Cellular_component
,	O
and	O
VPF	B-Gene_or_gene_product
/	O
VEGF	B-Gene_or_gene_product
also	O
induced	O
mRNA	O
encoding	O
osteopontin	B-Gene_or_gene_product
(	O
OPN	B-Gene_or_gene_product
)	O
,	O
an	O
alphavbeta3	B-Gene_or_gene_product
ligand	O
.	O

OPN	B-Gene_or_gene_product
promoted	O
EC	B-Cell
migration	O
in	O
vitro	O
;	O
and	O
VPF	B-Gene_or_gene_product
/	O
VEGF	B-Gene_or_gene_product
induction	O
of	O
alphavbeta3	B-Gene_or_gene_product
was	O
accompanied	O
by	O
increased	O
EC	B-Cell
migration	O
toward	O
OPN	B-Gene_or_gene_product
.	O

Because	O
thrombin	B-Gene_or_gene_product
cleavage	O
of	O
OPN	B-Gene_or_gene_product
results	O
in	O
substantial	O
enhancement	O
of	O
OPN	B-Gene_or_gene_product
'	O
s	O
adhesive	O
properties	O
,	O
and	O
because	O
VPF	B-Gene_or_gene_product
/	O
VEGF	B-Gene_or_gene_product
promotes	O
increased	O
microvascular	B-Tissue
permeability	O
leading	O
to	O
activation	O
of	O
the	O
extrinsic	O
coagulation	O
pathway	O
,	O
we	O
also	O
investigated	O
whether	O
VPF	B-Gene_or_gene_product
/	O
VEGF	B-Gene_or_gene_product
facilitates	O
thrombin	B-Gene_or_gene_product
cleavage	O
of	O
OPN	B-Gene_or_gene_product
in	O
vivo	O
.	O

Consistent	O
with	O
this	O
hypothesis	O
,	O
co	O
-	O
injection	O
of	O
VPF	B-Gene_or_gene_product
/	O
VEGF	B-Gene_or_gene_product
together	O
with	O
OPN	B-Gene_or_gene_product
resulted	O
in	O
rapid	O
cleavage	O
of	O
OPN	B-Gene_or_gene_product
by	O
endogenous	O
thrombin	B-Gene_or_gene_product
.	O

Furthermore	O
,	O
in	O
comparison	O
with	O
native	O
OPN	B-Gene_or_gene_product
,	O
thrombin	B-Gene_or_gene_product
-	O
cleaved	O
OPN	B-Gene_or_gene_product
stimulated	O
a	O
greater	O
rate	O
of	O
EC	B-Cell
migration	O
in	O
vitro	O
,	O
which	O
was	O
additive	O
to	O
the	O
increased	O
migration	O
associated	O
with	O
induction	O
of	O
alpha	B-Gene_or_gene_product
v	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
3	I-Gene_or_gene_product
.	O

Thus	O
,	O
these	O
data	O
demonstrate	O
cooperative	O
mechanisms	O
for	O
VPF	B-Gene_or_gene_product
/	O
VEGF	B-Gene_or_gene_product
regulation	O
of	O
EC	B-Cell
migration	O
involving	O
the	O
alphavbeta3	B-Gene_or_gene_product
integrin	I-Gene_or_gene_product
,	O
the	O
alphavbeta3	B-Gene_or_gene_product
ligand	O
OPN	B-Gene_or_gene_product
,	O
and	O
thrombin	B-Gene_or_gene_product
cleavage	O
of	O
OPN	B-Gene_or_gene_product
.	O

These	O
findings	O
also	O
illustrate	O
an	O
operational	O
link	O
between	O
VPF	B-Gene_or_gene_product
/	O
VEGF	B-Gene_or_gene_product
induction	O
of	O
EC	B-Cell
gene	O
expression	O
and	O
VPF	B-Gene_or_gene_product
/	O
VEGF	B-Gene_or_gene_product
enhancement	O
of	O
microvascular	B-Tissue
permeability	O
,	O
suggesting	O
that	O
these	O
distinct	O
biological	O
activities	O
may	O
act	O
accordingly	O
to	O
stimulate	O
EC	B-Cell
migration	O
during	O
angiogenesis	O
.	O

c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
erb	I-Gene_or_gene_product
B2	I-Gene_or_gene_product
overexpression	O
decreases	O
the	O
benefit	O
of	O
adjuvant	O
tamoxifen	B-Simple_chemical
in	O
early	B-Cancer
-	I-Cancer
stage	I-Cancer
breast	I-Cancer
cancer	I-Cancer
without	O
axillary	B-Multi-tissue_structure
lymph	I-Multi-tissue_structure
node	I-Multi-tissue_structure
metastases	O
.	O

PURPOSE	O
:	O
We	O
studied	O
retrospectively	O
the	O
interaction	O
between	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
erbB2	I-Gene_or_gene_product
overexpression	O
and	O
adjuvant	O
tomoxifen	B-Simple_chemical
in	O
node	B-Multi-tissue_structure
-	O
negative	O
breast	B-Cancer
cancer	I-Cancer
patients	B-Organism
enrolled	O
in	O
the	O
Gruppo	O
Universitario	O
Napoletano	O
1	O
(	O
GUN	O
-	O
1	O
)	O
trial	O
.	O

PATIENTS	O
AND	O
METHODS	O
:	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
erbB2	I-Gene_or_gene_product
,	O
evaluated	O
by	O
immunohistochemistry	O
in	O
145	O
of	O
173	O
patients	B-Organism
randomly	O
assigned	O
to	O
2	O
-	O
year	O
adjuvant	O
tamoxifen	B-Simple_chemical
or	O
no	O
further	O
therapy	O
,	O
was	O
considered	O
overexpressed	O
if	O
greater	O
than	O
10	O
%	O
of	O
the	O
cells	B-Cell
showed	O
specific	O
membrane	B-Cellular_component
staining	O
.	O

The	O
role	O
of	O
each	O
prognostic	O
variable	O
and	O
their	O
independent	O
effect	O
were	O
studied	O
using	O
the	O
Cox	O
model	O
.	O

Disease	O
-	O
free	O
(	O
DFS	O
)	O
and	O
overall	O
(	O
OAS	O
)	O
survival	O
curves	O
were	O
estimated	O
by	O
the	O
Kaplan	O
-	O
Meier	O
method	O
.	O

RESULTS	O
:	O
As	O
of	O
November	O
30	O
,	O
1994	O
,	O
the	O
median	O
follow	O
-	O
up	O
period	O
was	O
12	O
years	O
.	O

c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
erbB2	I-Gene_or_gene_product
was	O
overexpressed	O
in	O
43	O
of	O
145	O
patients	B-Organism
(	O
29	O
.	O
7	O
%	O
)	O
,	O
which	O
directly	O
correlated	O
with	O
tumor	B-Cancer
size	O
and	O
inversely	O
with	O
estrogen	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
(	O
ER	B-Gene_or_gene_product
)	O
level	O
.	O

At	O
univariate	O
analysis	O
,	O
overexpression	O
of	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
erbB2	I-Gene_or_gene_product
did	O
not	O
affect	O
either	O
DFS	O
or	O
OAS	O
;	O
tamoxifen	B-Simple_chemical
had	O
a	O
greater	O
effect	O
on	O
reducing	O
the	O
risk	O
of	O
recurrence	O
than	O
of	O
death	O
.	O

Addition	O
of	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
erbB2	I-Gene_or_gene_product
to	O
a	O
multivariate	O
Cox	O
model	O
that	O
contained	O
menopausal	O
status	O
,	O
tumor	B-Cancer
size	O
,	O
nuclear	B-Cellular_component
grade	O
,	O
and	O
treatment	O
as	O
covariates	O
did	O
not	O
affect	O
the	O
significance	O
of	O
the	O
model	O
for	O
DSF	O
or	O
OAS	O
,	O
whereas	O
addition	O
of	O
the	O
first	O
-	O
order	O
interaction	O
between	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
erbB2	I-Gene_or_gene_product
and	O
tamoxifen	B-Simple_chemical
was	O
statistically	O
significant	O
both	O
for	O
DFS	O
and	O
OAS	O
.	O

The	O
same	O
result	O
was	O
obtained	O
when	O
the	O
model	O
contained	O
ER	B-Gene_or_gene_product
status	O
and	O
ER	B-Gene_or_gene_product
-	O
tamoxifen	B-Simple_chemical
interaction	O
.	O

Indeed	O
,	O
adjuvant	O
tamoxifen	B-Simple_chemical
significantly	O
prolonged	O
DFS	O
and	O
OAS	O
in	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
erbB2	I-Gene_or_gene_product
-	O
negative	O
cases	O
,	O
whereas	O
it	O
had	O
no	O
effect	O
on	O
DFS	O
and	O
OAS	O
in	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
erbB2	I-Gene_or_gene_product
-	O
positive	O
patients	B-Organism
.	O

CONCLUSION	O
:	O
In	O
early	O
-	O
stage	O
breast	B-Cancer
cancer	I-Cancer
patients	B-Organism
,	O
overexpression	O
of	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
erbB2	I-Gene_or_gene_product
is	O
a	O
marker	O
of	O
lack	O
of	O
efficacy	O
of	O
adjuvant	O
tamoxifen	B-Simple_chemical
.	O

Gene	O
transfer	O
of	O
naked	O
DNA	B-Cellular_component
encoding	O
for	O
three	O
isoforms	O
of	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
stimulates	O
collateral	B-Multi-tissue_structure
development	O
in	O
vivo	O
.	O

Vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
is	O
a	O
naturally	O
secreted	O
endothelial	B-Cell
cell	I-Cell
-	O
specific	O
mitogen	O
.	O

We	O
investigated	O
the	O
hypothesis	O
that	O
naked	O
DNA	B-Cellular_component
encoding	O
for	O
VEGF	B-Gene_or_gene_product
could	O
be	O
used	O
in	O
a	O
strategy	O
of	O
arterial	B-Multi-tissue_structure
gene	O
therapy	O
to	O
stimulate	O
collateral	B-Multi-tissue_structure
artery	I-Multi-tissue_structure
development	O
.	O

Plasmid	B-Cellular_component
DNA	I-Cellular_component
encoding	O
each	O
of	O
the	O
three	O
principal	O
human	B-Organism
VEGF	B-Gene_or_gene_product
isoforms	O
(	O
phVEGF121	B-Gene_or_gene_product
,	O
phVEGF165	B-Gene_or_gene_product
,	O
or	O
phVEGF189	B-Gene_or_gene_product
)	O
was	O
applied	O
to	O
the	O
hydrogel	O
polymer	O
coating	O
of	O
an	O
angioplasty	O
balloon	O
and	O
delivered	O
percutaneously	O
to	O
one	O
iliac	B-Multi-tissue_structure
artery	I-Multi-tissue_structure
of	O
rabbits	B-Organism
with	O
operatively	O
induced	O
hindlimb	B-Organism_subdivision
ischemia	O
.	O

Compared	O
with	O
control	O
animals	O
transfected	O
with	O
LacZ	B-Gene_or_gene_product
,	O
site	O
-	O
specific	O
transfection	O
of	O
phVEGF	B-Gene_or_gene_product
resulted	O
in	O
augmented	O
collateral	B-Multi-tissue_structure
vessel	I-Multi-tissue_structure
development	O
documented	O
by	O
serial	O
angiography	O
,	O
and	O
improvement	O
in	O
calf	B-Organism_subdivision
blood	B-Organism_substance
pressure	O
ratio	O
(	O
ischemic	B-Organism_subdivision
to	O
normal	B-Organism_subdivision
limb	I-Organism_subdivision
)	O
,	O
resting	O
and	O
maximum	O
blood	B-Organism_substance
flow	O
,	O
and	O
capillary	B-Tissue
to	O
myocyte	B-Cell
ratio	O
.	O

Similar	O
results	O
were	O
obtained	O
with	O
phVEGF121	B-Gene_or_gene_product
,	O
phVEGF165	B-Gene_or_gene_product
,	O
and	O
phVEGF189	B-Gene_or_gene_product
,	O
which	O
suggests	O
that	O
these	O
isoforms	O
are	O
biologically	O
equivalent	O
with	O
respect	O
to	O
in	O
vivo	O
angiogenesis	O
.	O

The	O
fact	O
that	O
viral	O
or	O
other	O
adjunctive	O
vectors	O
were	O
not	O
required	O
further	O
suggests	O
that	O
secreted	O
gene	O
products	O
may	O
have	O
potential	O
therapeutic	O
utility	O
even	O
when	O
the	O
number	O
of	O
successfully	O
transfected	O
cells	B-Cell
remains	O
low	O
.	O

Arterial	B-Multi-tissue_structure
gene	O
transfer	O
of	O
naked	O
DNA	B-Cellular_component
encoding	O
for	O
a	O
secreted	O
angiogenic	O
cytokine	O
,	O
thus	O
,	O
represents	O
a	O
potential	O
alternative	O
to	O
recombinant	O
protein	O
administration	O
for	O
stimulating	O
collateral	B-Multi-tissue_structure
vessel	I-Multi-tissue_structure
development	O
.	O

Comparison	O
of	O
transformation	O
by	O
manganese	B-Simple_chemical
sulfate	I-Simple_chemical
and	O
5	B-Simple_chemical
-	I-Simple_chemical
azacytidine	I-Simple_chemical
in	O
Rat	B-Cell
6	I-Cell
cells	I-Cell
overexpressing	O
the	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
oncogene	O
.	O

Rat	B-Cell
6	I-Cell
cells	I-Cell
are	O
not	O
transformed	O
by	O
treatment	O
with	O
the	O
well	O
-	O
known	O
carcinogens	O
benzo	B-Simple_chemical
[	I-Simple_chemical
a	I-Simple_chemical
]	I-Simple_chemical
pyrene	I-Simple_chemical
(	O
BP	B-Simple_chemical
)	O
or	O
N	B-Simple_chemical
-	I-Simple_chemical
methyl	I-Simple_chemical
-	I-Simple_chemical
N	I-Simple_chemical
-	I-Simple_chemical
nitro	I-Simple_chemical
-	I-Simple_chemical
N	I-Simple_chemical
'	I-Simple_chemical
-	I-Simple_chemical
nitrosoguanidine	I-Simple_chemical
(	O
MNNG	B-Simple_chemical
)	O
.	O

Upon	O
retroviral	B-Organism
transduction	O
of	O
the	O
mouse	B-Organism
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
gene	O
,	O
Rat	B-Cell
6	I-Cell
cells	I-Cell
showed	O
mildly	O
altered	O
morphology	O
and	O
formed	O
microcolonies	O
in	O
soft	O
agar	O
;	O
furthermore	O
,	O
they	O
could	O
be	O
transformed	O
by	O
BP	B-Simple_chemical
and	O
MNNG	B-Simple_chemical
to	O
form	O
large	O
colonies	O
in	O
agar	O
(	O
Hsiao	O
et	O
al	O
.	O
(	O
1992	O
)	O
Mol	O
.	O
Carcinogenesis	O
,	O
5	O
,	O
140	O
-	O
154	O
)	O
.	O

In	O
the	O
current	O
report	O
,	O
we	O
tested	O
the	O
sensitivity	O
of	O
the	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
-	O
overexpressing	O
cells	O
(	O
Rat	O
6	O
/	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
)	O
to	O
two	O
additional	O
chemicals	O
:	O
5	B-Simple_chemical
-	I-Simple_chemical
azacytidine	I-Simple_chemical
and	O
MnSO4	B-Simple_chemical
.	O

These	O
chemicals	O
differ	O
from	O
the	O
direct	O
-	O
acting	O
mutagens	O
tested	O
previously	O
.	O

5	B-Simple_chemical
-	I-Simple_chemical
Azacytidine	I-Simple_chemical
,	O
a	O
potent	O
DNA	B-Cellular_component
methylation	O
inhibitor	O
,	O
induced	O
growth	O
of	O
large	O
colonies	O
in	O
soft	O
agar	O
cultures	O
of	O
Rat	B-Cell
6	I-Cell
or	O
Rat	O
6	O
/	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
cells	O
.	O

On	O
the	O
other	O
hand	O
,	O
MnSO4	B-Simple_chemical
only	O
induced	O
transformation	O
in	O
Rat	O
6	O
/	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
cells	O
,	O
but	O
not	O
the	O
parental	O
Rat	B-Cell
6	I-Cell
cells	I-Cell
.	O

Transformants	B-Cell
induced	O
by	O
5	B-Simple_chemical
-	I-Simple_chemical
azacytidine	I-Simple_chemical
lost	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
-	O
induced	O
apoptotic	O
cell	B-Cell
death	O
,	O
whereas	O
MnSO4	B-Simple_chemical
-	O
induced	O
transformants	B-Cell
showed	O
a	O
higher	O
degree	O
of	O
apoptosis	O
than	O
the	O
parental	O
Rat	O
6	O
/	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
cells	O
.	O

These	O
results	O
suggest	O
that	O
MnSO4	B-Simple_chemical
co	O
-	O
operates	O
with	O
overexpressed	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
in	O
inducing	O
transformation	O
,	O
while	O
5	B-Simple_chemical
-	I-Simple_chemical
azacytidine	I-Simple_chemical
transformation	O
is	O
independent	O
of	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
overexpression	O
and	O
may	O
involve	O
alterations	O
in	O
the	O
regulation	O
of	O
apoptosis	O
.	O

Regulation	O
of	O
p53	B-Gene_or_gene_product
-	O
dependent	O
apoptosis	O
,	O
transcriptional	O
repression	O
,	O
and	O
cell	B-Cell
transformation	O
by	O
phosphorylation	O
of	O
the	O
55	O
-	O
kilodalton	O
E1B	B-Gene_or_gene_product
protein	O
of	O
human	B-Organism
adenovirus	I-Organism
type	I-Organism
5	I-Organism
.	O

The	O
adenovirus	B-Organism
type	I-Organism
5	I-Organism
55	O
-	O
kDa	O
E1B	B-Gene_or_gene_product
protein	O
(	O
E1B	B-Gene_or_gene_product
-	O
55kDa	O
)	O
cooperates	O
with	O
E1A	B-Gene_or_gene_product
gene	O
products	O
to	O
induce	O
cell	B-Cell
transformation	O
.	O

E1A	B-Gene_or_gene_product
proteins	O
stimulate	O
DNA	B-Cellular_component
synthesis	O
and	O
cell	B-Cell
proliferation	O
;	O
however	O
,	O
they	O
also	O
cause	O
rapid	O
cell	B-Cell
death	O
by	O
p53	B-Gene_or_gene_product
-	O
dependent	O
and	O
p53	B-Gene_or_gene_product
-	O
independent	O
apoptosis	O
.	O

It	O
is	O
believed	O
that	O
the	O
role	O
of	O
the	O
E1B	B-Gene_or_gene_product
-	O
55kDa	O
protein	O
in	O
transformation	O
is	O
to	O
protect	O
against	O
p53	B-Gene_or_gene_product
-	O
dependent	O
apoptosis	O
by	O
binding	O
to	O
and	O
inactivating	O
p53	B-Gene_or_gene_product
.	O

It	O
has	O
been	O
shown	O
previously	O
that	O
the	O
55	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kDa	I-Gene_or_gene_product
polypeptide	O
abrogates	O
p53	B-Gene_or_gene_product
-	O
mediated	O
transactivation	O
and	O
that	O
mutants	O
defective	O
in	O
p53	B-Gene_or_gene_product
binding	O
are	O
unable	O
to	O
cooperate	O
with	O
E1A	B-Gene_or_gene_product
in	O
transformation	O
.	O

We	O
have	O
previously	O
mapped	O
phosphorylation	O
sites	O
near	O
the	O
carboxy	O
terminus	O
of	O
the	O
E1B	B-Gene_or_gene_product
-	O
55kDa	O
protein	O
at	O
Ser	B-Amino_acid
-	I-Amino_acid
490	I-Amino_acid
and	O
Ser	B-Amino_acid
-	I-Amino_acid
491	I-Amino_acid
,	O
which	O
lie	O
within	O
casein	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
II	I-Gene_or_gene_product
consensus	O
sequences	O
.	O

Conversion	O
of	O
these	O
sites	O
to	O
alanine	B-Amino_acid
residues	O
greatly	O
reduced	O
transforming	O
activity	O
,	O
and	O
although	O
the	O
mutant	O
55	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kDa	I-Gene_or_gene_product
protein	O
was	O
found	O
to	O
interact	O
with	O
p53	B-Gene_or_gene_product
at	O
normal	O
levels	O
,	O
it	O
was	O
somewhat	O
defective	O
for	O
suppression	O
of	O
p53	B-Gene_or_gene_product
transactivation	O
activity	O
.	O

We	O
now	O
report	O
that	O
a	O
nearby	O
residue	O
,	O
Thr	B-Amino_acid
-	I-Amino_acid
495	I-Amino_acid
,	O
also	O
appears	O
to	O
be	O
phosphorylated	O
.	O

We	O
demonstrate	O
directly	O
that	O
the	O
wild	O
-	O
type	O
55	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kDa	I-Gene_or_gene_product
protein	O
is	O
able	O
to	O
block	O
E1A	B-Gene_or_gene_product
-	O
induced	O
p53	B-Gene_or_gene_product
-	O
dependent	O
apoptosis	O
,	O
whereas	O
cells	B-Cell
infected	O
by	O
mutant	B-Organism
pm490	I-Organism
/	I-Organism
1	I-Organism
/	I-Organism
5A	I-Organism
,	O
which	O
contains	O
alanine	B-Amino_acid
residues	O
at	O
all	O
three	O
phosphorylation	O
sites	O
,	O
exhibited	O
extensive	O
DNA	B-Cellular_component
fragmentation	O
and	O
classic	O
apoptotic	O
cell	B-Cell
death	O
.	O

The	O
E1B	B-Gene_or_gene_product
-	O
55kDa	O
product	O
has	O
been	O
shown	O
to	O
exhibit	O
intrinsic	O
transcriptional	O
repression	O
activity	O
when	O
localized	O
to	O
promoters	O
,	O
such	O
as	O
by	O
fusion	O
with	O
the	O
GAL4	B-Gene_or_gene_product
DNA	B-Cellular_component
-	O
binding	O
domain	O
,	O
even	O
in	O
the	O
absence	O
of	O
p53	B-Gene_or_gene_product
.	O

Such	O
repression	O
activity	O
was	O
totally	O
absent	O
with	O
mutant	O
pm490	O
/	O
1	O
/	O
5A	O
.	O

These	O
data	O
suggested	O
that	O
inhibition	O
of	O
p53	B-Gene_or_gene_product
-	O
dependent	O
apoptosis	O
may	O
depend	O
on	O
the	O
transcriptional	O
repression	O
function	O
of	O
the	O
55	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kDa	I-Gene_or_gene_product
protein	O
,	O
which	O
appears	O
to	O
be	O
regulated	O
be	O
phosphorylation	O
at	O
the	O
carboxy	O
terminus	O
.	O

Non	O
-	O
invasive	O
grading	O
of	O
primary	O
brain	B-Cancer
tumours	I-Cancer
:	O
results	O
of	O
a	O
comparative	O
study	O
between	O
SPET	O
with	O
123I	B-Simple_chemical
-	I-Simple_chemical
alpha	I-Simple_chemical
-	I-Simple_chemical
methyl	I-Simple_chemical
tyrosine	I-Simple_chemical
and	O
PET	O
with	O
18F	B-Simple_chemical
-	I-Simple_chemical
deoxyglucose	I-Simple_chemical
.	O

Use	O
of	O
iodine	B-Simple_chemical
-	I-Simple_chemical
123	I-Simple_chemical
-	I-Simple_chemical
alpha	I-Simple_chemical
-	I-Simple_chemical
methyl	I-Simple_chemical
tyrosine	I-Simple_chemical
(	O
123I	B-Simple_chemical
-	I-Simple_chemical
IMT	I-Simple_chemical
)	O
allows	O
investigation	O
of	O
the	O
amino	B-Amino_acid
acid	I-Amino_acid
transport	O
rate	O
in	O
gliomas	B-Cancer
.	O

It	O
was	O
the	O
aim	O
of	O
this	O
study	O
to	O
compare	O
the	O
value	O
of	O
measurement	O
of	O
glucose	B-Simple_chemical
metabolism	O
with	O
that	O
of	O
measurement	O
of	O
123I	B-Simple_chemical
-	I-Simple_chemical
IMT	I-Simple_chemical
uptake	O
for	O
the	O
non	O
-	O
invasive	O
grading	O
of	O
brain	B-Cancer
tumours	I-Cancer
.	O

The	O
study	O
population	O
comprised	O
23	O
patients	B-Organism
with	O
histopathologically	O
proven	O
primary	O
brain	B-Cancer
tumours	I-Cancer
;	O
14	O
had	O
high	B-Cancer
-	I-Cancer
grade	I-Cancer
gliomas	I-Cancer
,	O
and	O
nine	O
low	B-Cancer
-	I-Cancer
grade	I-Cancer
brain	I-Cancer
neoplasms	I-Cancer
.	O

Glucose	B-Simple_chemical
metabolism	O
was	O
studied	O
using	O
an	O
ECAT	O
EXACT	O
47	O
positron	O
emission	O
tomography	O
(	O
PET	O
)	O
camera	O
and	O
fluorine	B-Simple_chemical
-	I-Simple_chemical
18	I-Simple_chemical
fluorodeoxyglucose	I-Simple_chemical
(	O
18F	B-Simple_chemical
-	I-Simple_chemical
FDG	I-Simple_chemical
)	O
;	O
123I	B-Simple_chemical
-	I-Simple_chemical
IMT	I-Simple_chemical
uptake	O
was	O
measured	O
with	O
the	O
triple	O
-	O
headed	O
single	O
-	O
photon	O
emission	O
tomography	O
(	O
SPET	O
)	O
camera	O
,	O
MULTISPECT	O
3	O
.	O

18F	B-Simple_chemical
-	I-Simple_chemical
FDG	I-Simple_chemical
and	O
123I	B-Simple_chemical
-	I-Simple_chemical
IMT	I-Simple_chemical
uptake	O
was	O
quantified	O
as	O
ratios	O
between	O
the	O
uptake	O
by	O
the	O
tumour	B-Cancer
and	O
contralateral	O
regions	O
of	O
reference	O
.	O

Glucose	B-Simple_chemical
metabolism	O
and	O
amino	B-Amino_acid
acid	I-Amino_acid
uptake	O
of	O
the	O
brain	B-Cancer
tumours	I-Cancer
correlated	O
significantly	O
(	O
r	O
=	O
0	O
.	O
71	O
,	O
P	O
<	O
0	O
.	O
001	O
)	O
.	O

Assuming	O
discrimination	O
thresholds	O
between	O
high	B-Cancer
-	I-Cancer
grade	I-Cancer
and	O
low	B-Cancer
-	I-Cancer
grade	I-Cancer
tumours	I-Cancer
of	O
0	O
.	O
8	O
for	O
18F	B-Simple_chemical
-	I-Simple_chemical
FDG	I-Simple_chemical
uptake	O
and	O
1	O
.	O
8	O
for	O
123I	B-Simple_chemical
-	I-Simple_chemical
IMT	I-Simple_chemical
uptake	O
,	O
the	O
accuracy	O
values	O
of	O
18F	B-Simple_chemical
-	I-Simple_chemical
FDG	I-Simple_chemical
PET	O
and	O
123I	B-Simple_chemical
-	I-Simple_chemical
IMT	I-Simple_chemical
SPET	O
for	O
differentiating	O
between	O
high	B-Cancer
-	I-Cancer
grade	I-Cancer
and	O
low	B-Cancer
-	I-Cancer
grade	I-Cancer
tumours	I-Cancer
were	O
21	O
/	O
23	O
(	O
91	O
%	O
)	O
and	O
19	O
/	O
23	O
(	O
83	O
%	O
)	O
,	O
respectively	O
.	O

The	O
difference	O
in	O
diagnostic	O
performance	O
was	O
not	O
significant	O
on	O
receiver	O
operating	O
characteristic	O
analysis	O
(	O
P	O
>	O
0	O
.	O
4	O
)	O
.	O

It	O
is	O
concluded	O
that	O
there	O
is	O
no	O
major	O
difference	O
between	O
the	O
PET	O
investigation	O
of	O
glucose	B-Simple_chemical
metabolism	O
and	O
the	O
less	O
expensive	O
SPET	O
measurement	O
of	O
amino	B-Amino_acid
acid	I-Amino_acid
uptake	O
in	O
terms	O
of	O
their	O
accuracy	O
in	O
evaluating	O
the	O
malignancy	O
grade	O
of	O
primary	O
brain	B-Cancer
tumours	I-Cancer
.	O

This	O
encourages	O
the	O
performance	O
of	O
further	O
studies	O
to	O
analyse	O
the	O
potential	O
impact	O
of	O
123I	B-Simple_chemical
-	I-Simple_chemical
IMT	I-Simple_chemical
SPET	O
on	O
the	O
therapeutic	O
management	O
of	O
patients	B-Organism
with	O
brain	B-Cancer
tumours	I-Cancer
.	O

The	O
Wilms	B-Cancer
tumor	I-Cancer
suppressor	O
gene	O
WT1	B-Gene_or_gene_product
induces	O
G1	O
arrest	O
and	O
apoptosis	O
in	O
myeloblastic	B-Cell
leukemia	I-Cell
M1	I-Cell
cells	I-Cell
.	O

WT1	B-Gene_or_gene_product
was	O
isolated	O
as	O
a	O
tumor	B-Cancer
suppressor	O
gene	O
of	O
Wilms	B-Cancer
tumor	I-Cancer
.	O

However	O
,	O
high	O
expression	O
of	O
WT1	B-Gene_or_gene_product
correlates	O
with	O
poor	O
prognosis	O
in	O
acute	B-Cancer
leukemia	I-Cancer
.	O

In	O
addition	O
suppression	O
of	O
WT1	B-Gene_or_gene_product
expression	O
by	O
WT1	B-Gene_or_gene_product
anti	O
-	O
sense	O
oligonucleotide	O
inhibits	O
proliferation	O
of	O
leukemia	B-Cell
cells	I-Cell
,	O
suggesting	O
that	O
WT1	B-Gene_or_gene_product
is	O
important	O
for	O
their	O
proliferation	O
.	O

To	O
further	O
elucidate	O
the	O
biological	O
significance	O
of	O
WT1	B-Gene_or_gene_product
in	O
leukemic	O
cell	B-Cell
growth	O
,	O
we	O
overexpressed	O
exogenous	O
WT1	B-Gene_or_gene_product
in	O
murine	B-Organism
M1	B-Cell
myeloblastic	I-Cell
leukemia	I-Cell
cells	I-Cell
using	O
the	O
isopropyl	B-Simple_chemical
-	I-Simple_chemical
beta	I-Simple_chemical
-	I-Simple_chemical
D	I-Simple_chemical
-	I-Simple_chemical
thiogalactoside	I-Simple_chemical
(	O
IPTG	B-Simple_chemical
)	O
-	O
controlled	O
expression	O
system	O
.	O

We	O
found	O
that	O
induction	O
of	O
one	O
splicing	O
variant	O
of	O
WT1	B-Gene_or_gene_product
[	O
WT1	B-Gene_or_gene_product
-	I-Gene_or_gene_product
17AA	I-Gene_or_gene_product
(	I-Gene_or_gene_product
+	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	I-Gene_or_gene_product
KTS	I-Gene_or_gene_product
(	I-Gene_or_gene_product
-	I-Gene_or_gene_product
)	I-Gene_or_gene_product
]	O
in	O
M1	B-Cell
cells	I-Cell
induces	O
cell	B-Cell
cycle	O
arrest	O
and	O
apoptotic	O
cell	B-Cell
death	O
.	O

These	O
results	O
suggest	O
that	O
the	O
role	O
of	O
WT1	B-Gene_or_gene_product
is	O
different	O
depending	O
on	O
the	O
type	O
of	O
leukemia	B-Cell
cell	I-Cell
in	O
which	O
it	O
is	O
expressed	O
.	O

[	O
Expression	O
of	O
metastasis	O
suppressor	O
gene	O
nm23	B-Gene_or_gene_product
in	O
human	B-Organism
hepatocellular	B-Cancer
carcinoma	I-Cancer
]	O
.	O

For	O
the	O
purpose	O
of	O
investigating	O
the	O
relationship	O
between	O
the	O
metastatic	O
potential	O
of	O
the	O
tumor	B-Cancer
as	O
well	O
as	O
the	O
expression	O
of	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
mRNA	O
,	O
and	O
for	O
determing	O
the	O
location	O
of	O
the	O
positive	O
sites	O
in	O
the	O
cells	B-Cell
,	O
tumor	B-Cancer
metastasis	O
suppressor	O
gene	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
in	O
human	B-Organism
hepatocellular	B-Cancer
carcinoma	I-Cancer
(	O
and	O
the	O
nonneoplastic	B-Multi-tissue_structure
area	I-Multi-tissue_structure
surrounding	O
the	O
tumor	B-Cancer
)	O
was	O
detected	O
by	O
in	O
situ	O
hybridization	O
using	O
digoxiginin	B-Simple_chemical
-	O
labeled	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
antisense	O
complementary	O
RNA	O
probe	O
.	O

The	O
primary	O
results	O
indicated	O
(	O
i	O
)	O
positive	O
results	O
of	O
in	O
situ	O
hybridization	O
are	O
presence	O
of	O
granules	B-Cellular_component
or	O
masses	O
in	O
the	O
cytoplasm	B-Organism_substance
;	O
(	O
ii	O
)	O
the	O
less	O
expression	O
of	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
mRNA	O
,	O
the	O
higher	O
metastatic	O
rate	O
of	O
the	O
human	B-Organism
hepatocellular	B-Cancer
carcinoma	I-Cancer
(	O
P	O
<	O
0	O
.	O
05	O
)	O
;	O
(	O
iii	O
)	O
expression	O
of	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
mRNA	O
dose	O
not	O
correlate	O
with	O
some	O
other	O
factors	O
such	O
as	O
tumor	B-Cancer
size	O
and	O
the	O
background	O
of	O
other	O
liver	B-Organ
diseases	O
.	O

CD56	B-Gene_or_gene_product
+	O
putative	O
natural	B-Cancer
killer	I-Cancer
cell	I-Cancer
lymphomas	I-Cancer
:	O
production	O
of	O
cytolytic	O
effectors	O
and	O
related	O
proteins	O
mediating	O
tumor	B-Cell
cell	I-Cell
apoptosis	O
?	O

Apoptosis	O
is	O
a	O
regulated	O
form	O
of	O
cell	B-Cell
death	O
that	O
may	O
be	O
triggered	O
by	O
natural	B-Cell
killer	I-Cell
(	O
NK	B-Cell
)	O
or	O
cytotoxic	B-Cell
T	I-Cell
cells	I-Cell
,	O
which	O
effect	O
target	O
cell	B-Cell
lysis	O
by	O
cytolytic	O
effector	O
and	O
related	O
proteins	O
through	O
complex	O
intracellular	B-Immaterial_anatomical_entity
signals	O
.	O

This	O
study	O
was	O
aimed	O
to	O
investigate	O
whether	O
there	O
is	O
selective	O
expression	O
of	O
these	O
cytolytic	O
markers	O
in	O
the	O
putative	O
NK	B-Cancer
-	I-Cancer
cell	I-Cancer
lymphomas	I-Cancer
and	O
whether	O
there	O
is	O
correlation	O
with	O
zonal	O
tumor	B-Cell
cell	I-Cell
death	O
in	O
these	O
tumors	B-Cancer
.	O

Expression	O
of	O
the	O
cytolytic	O
effectors	O
perforin	B-Gene_or_gene_product
,	O
granzyme	B-Gene_or_gene_product
B9	I-Gene_or_gene_product
,	O
and	O
the	O
granule	B-Cellular_component
membrane	I-Cellular_component
protein	O
TIA1	B-Gene_or_gene_product
were	O
examined	O
in	O
24	O
putative	O
NK	B-Cancer
-	I-Cancer
cell	I-Cancer
lymphomas	I-Cancer
,	O
18	O
postthymic	O
T	B-Cancer
-	I-Cancer
cell	I-Cancer
lymphomas	I-Cancer
(	O
one	O
case	O
CD8	B-Gene_or_gene_product
+	O
CD56	B-Gene_or_gene_product
+	O
and	O
three	O
anaplastic	B-Cancer
large	I-Cancer
cell	I-Cancer
lymphomas	I-Cancer
(	O
ALCL	B-Cancer
)	O
,	O
three	O
T	B-Cancer
-	I-Cancer
lymphoblastic	I-Cancer
lymphomas	I-Cancer
,	O
and	O
20	O
B	B-Cancer
-	I-Cancer
cell	I-Cancer
lymphomas	I-Cancer
.	O

Nineteen	O
(	O
79	O
%	O
)	O
putative	O
NK	B-Cancer
-	I-Cancer
cell	I-Cancer
lymphomas	I-Cancer
expressed	O
perforin	B-Gene_or_gene_product
,	O
and	O
all	O
24	O
cases	O
expressed	O
granzyme	B-Gene_or_gene_product
B9	I-Gene_or_gene_product
and	O
TIA1	B-Gene_or_gene_product
.	O

The	O
only	O
CD8	B-Gene_or_gene_product
+	O
CD56	B-Gene_or_gene_product
+	O
postthymic	O
T	O
-	O
cell	O
lymphoma	O
also	O
expressed	O
all	O
three	O
cytolytic	O
markers	O
,	O
two	O
CD8	B-Gene_or_gene_product
-	O
ALCL	O
expressed	O
TIA1	B-Gene_or_gene_product
;	O
other	O
postthymic	B-Cancer
T	I-Cancer
-	I-Cancer
cell	I-Cancer
,	O
T	B-Cancer
-	I-Cancer
lymphoblastic	I-Cancer
,	O
and	O
B	B-Cancer
-	I-Cancer
cell	I-Cancer
lymphomas	I-Cancer
were	O
consistently	O
negative	O
.	O

There	O
was	O
strong	O
correlation	O
between	O
percentage	O
perforin	B-Gene_or_gene_product
-	O
positive	O
cells	B-Cell
and	O
zonal	B-Cell
tumor	I-Cell
cell	I-Cell
death	O
.	O

Angioinvasion	O
,	O
in	O
contrast	O
,	O
was	O
present	O
only	O
in	O
a	O
proportion	O
(	O
37	O
%	O
)	O
of	O
these	O
lymphomas	B-Cancer
despite	O
the	O
frequent	O
presence	O
of	O
zonal	O
tumor	B-Cell
cell	I-Cell
death	O
(	O
71	O
%	O
)	O
.	O

We	O
propose	O
that	O
cytolytic	O
effector	O
and	O
related	O
proteins	O
produced	O
by	O
putative	O
NK	B-Cell
and	O
some	O
CD8	B-Gene_or_gene_product
+	O
CD56	B-Gene_or_gene_product
+	O
postthymic	O
T	O
-	O
cell	O
lymphomas	O
,	O
probably	O
in	O
conjunction	O
with	O
other	O
mechanisms	O
,	O
may	O
effect	O
massive	O
tumor	B-Cell
cell	I-Cell
apoptosis	O
.	O

The	O
frequent	O
expression	O
of	O
cytolytic	O
effector	O
markers	O
in	O
the	O
CD2	B-Gene_or_gene_product
+	O
surface	O
CD3	B-Gene_or_gene_product
-	O
CD56	B-Gene_or_gene_product
+	O
putative	O
NK	O
-	O
cell	O
lymphomas	O
lends	O
further	O
support	O
to	O
their	O
probable	O
NK	B-Cell
cell	I-Cell
origin	O
.	O

Prophylactic	O
thyroidectomy	O
in	O
MEN	B-Cancer
IIA	I-Cancer
:	O
does	O
the	O
calcitonin	B-Gene_or_gene_product
level	O
correlate	O
with	O
tumor	B-Cancer
spread	O
?	O

BACKGROUND	O
:	O
The	O
fate	O
of	O
patients	B-Organism
with	O
multiple	B-Cancer
endocrine	I-Cancer
neoplasia	I-Cancer
of	I-Cancer
type	I-Cancer
II	I-Cancer
A	I-Cancer
(	O
MEN	B-Cancer
II	I-Cancer
A	I-Cancer
)	O
is	O
determined	O
by	O
medullary	B-Cancer
thyroid	I-Cancer
carcinoma	I-Cancer
,	O
which	O
occurs	O
in	O
all	O
cases	O
.	O

This	O
has	O
led	O
to	O
the	O
therapeutic	O
concept	O
of	O
prophylactic	O
thyroidectomy	O
in	O
affected	O
family	O
members	O
with	O
the	O
goal	O
of	O
removing	O
the	O
thyroid	B-Organ
before	O
the	O
manifestation	O
of	O
carcinoma	B-Cancer
.	O

We	O
investigated	O
a	O
prophylactically	O
thyroidectomized	O
MEN	B-Cancer
II	I-Cancer
A	I-Cancer
population	O
to	O
determine	O
whether	O
the	O
highly	O
specific	O
and	O
sensitive	O
tumor	B-Cancer
marker	O
calcitonin	B-Gene_or_gene_product
correlates	O
with	O
tumor	B-Cancer
spread	O
.	O

PATIENTS	O
AND	O
METHODS	O
:	O
Fifteen	O
patients	B-Organism
with	O
MEN	B-Cancer
II	I-Cancer
A	I-Cancer
(	O
aged	O
4	O
-	O
24	O
years	O
)	O
who	O
had	O
undergone	O
prophylactic	O
thyroidectomy	O
since	O
1990	O
were	O
included	O
in	O
the	O
study	O
.	O

Baseline	O
and	O
pentagastrin	B-Simple_chemical
-	O
stimulated	O
calcitonin	B-Gene_or_gene_product
levels	O
were	O
preoperatively	O
determined	O
in	O
all	O
cases	O
.	O

The	O
indication	O
for	O
surgery	O
was	O
established	O
on	O
the	O
basis	O
of	O
pathologic	O
calcitonin	B-Gene_or_gene_product
levels	O
in	O
the	O
first	O
seven	O
patients	B-Organism
and	O
on	O
the	O
basis	O
of	O
detected	O
RET	B-Gene_or_gene_product
proto	O
-	O
oncogene	O
mutation	O
in	O
the	O
other	O
eight	O
patients	B-Organism
.	O

Bilateral	O
central	O
lymphadenectomy	O
was	O
performed	O
in	O
all	O
patients	B-Organism
in	O
addition	O
to	O
thyroidectomy	O
.	O

RESULTS	O
:	O
Histology	O
demonstrated	O
C	B-Cell
-	I-Cell
cell	I-Cell
hyperplasia	O
in	O
five	O
patients	B-Organism
(	O
aged	O
4	O
-	O
13	O
years	O
)	O
,	O
unilateral	O
medullary	B-Cancer
microcarcinoma	I-Cancer
in	O
six	O
(	O
aged	O
9	O
-	O
17	O
years	O
)	O
and	O
a	O
bilateral	O
medullary	B-Cancer
microcarcinoma	I-Cancer
in	O
three	O
cases	O
(	O
aged	O
17	O
-	O
24	O
years	O
)	O
.	O

One	O
9	O
-	O
year	O
-	O
old	O
boy	B-Organism
with	O
bilateral	O
microcarcinoma	B-Cancer
already	O
had	O
a	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
.	O

The	O
mean	O
baseline	O
calcitonin	B-Gene_or_gene_product
level	O
correlated	O
with	O
the	O
histologic	O
findings	O
(	O
r	O
=	O
0	O
.	O
71	O
,	O
P	O
=	O
0	O
.	O
003	O
)	O
but	O
there	O
was	O
no	O
correlation	O
between	O
pentagastrin	B-Simple_chemical
-	O
stimulated	O
calcitonin	B-Gene_or_gene_product
levels	O
and	O
histology	O
(	O
r	O
=	O
0	O
.	O
21	O
,	O
P	O
=	O
0	O
.	O
47	O
)	O
.	O

CONCLUSION	O
:	O
In	O
MEN	B-Cancer
II	I-Cancer
A	I-Cancer
patients	B-Organism
undergoing	O
prophylactic	O
thyroidectomy	O
,	O
baseline	O
but	O
not	O
stimulated	O
calcitonin	B-Gene_or_gene_product
levels	O
already	O
correlate	O
with	O
the	O
histologic	O
tumor	B-Cancer
stage	O
at	O
the	O
stage	O
of	O
clinically	O
occult	O
C	B-Cell
-	I-Cell
cell	I-Cell
hyperplasia	O
or	O
medullary	B-Cancer
microcarcinoma	I-Cancer
.	O

However	O
,	O
biochemical	O
screening	O
cannot	O
reliably	O
discriminate	O
the	O
transition	O
from	O
C	B-Cell
-	I-Cell
cell	I-Cell
hyperplasia	O
to	O
invasive	O
microcarcinoma	B-Cancer
.	O

Individuals	O
with	O
MEN	B-Cancer
IIA	I-Cancer
should	O
therefore	O
undergo	O
early	O
prophylactic	O
thyroidectomy	O
once	O
the	O
diagnosis	O
is	O
confirmed	O
by	O
molecular	O
genetic	O
testing	O
.	O

Induction	O
of	O
Ets	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
in	O
endothelial	B-Cell
cells	I-Cell
during	O
reendothelialization	O
after	O
denuding	O
injury	O
.	O

Ets	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
a	O
transcription	O
factor	O
,	O
is	O
induced	O
in	O
endothelial	B-Cell
cells	I-Cell
(	O
ECs	B-Cell
)	O
during	O
angiogenesis	O
.	O

Here	O
,	O
we	O
investigated	O
the	O
expression	O
of	O
Ets	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
during	O
reendothelialization	O
.	O

When	O
a	O
confluent	O
monolayer	B-Cell
of	O
human	B-Cell
umbilical	I-Cell
vein	I-Cell
endothelial	I-Cell
cell	I-Cell
line	I-Cell
,	O
ECV304	B-Cell
,	O
was	O
denuded	O
,	O
ECV304	B-Cell
at	O
the	O
wound	B-Pathological_formation
edge	I-Pathological_formation
expressed	O
Ets	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

An	O
immunohistochemical	O
analysis	O
revealed	O
that	O
Ets	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
accumulated	O
in	O
migrating	B-Cell
cells	I-Cell
at	O
the	O
wound	B-Pathological_formation
edge	I-Pathological_formation
and	O
returned	O
to	O
basal	O
level	O
when	O
reendothelialization	O
was	O
accomplished	O
.	O

This	O
induction	O
of	O
Ets	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
could	O
be	O
reproduced	O
in	O
in	O
vivo	O
denudation	O
of	O
rat	B-Organism
aortic	B-Tissue
endothelium	I-Tissue
by	O
a	O
balloon	O
catheter	O
.	O

The	O
induction	O
of	O
Ets	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
in	O
ECs	B-Cell
after	O
denudation	O
was	O
regulated	O
transcriptionally	O
,	O
and	O
humeral	B-Gene_or_gene_product
factors	I-Gene_or_gene_product
released	O
from	O
injured	O
ECs	B-Cell
might	O
not	O
be	O
responsible	O
.	O

Mitogen	B-Gene_or_gene_product
-	I-Gene_or_gene_product
activated	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
(	O
MAPK	B-Gene_or_gene_product
)	O
activities	O
were	O
investigated	O
to	O
explore	O
the	O
mechanism	O
of	O
this	O
induction	O
.	O

Although	O
extracellular	B-Gene_or_gene_product
signal	I-Gene_or_gene_product
-	I-Gene_or_gene_product
regulated	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
1	I-Gene_or_gene_product
/	O
2	B-Gene_or_gene_product
(	O
ERK1	B-Gene_or_gene_product
/	O
2	B-Gene_or_gene_product
)	O
,	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Jun	I-Gene_or_gene_product
N	I-Gene_or_gene_product
-	I-Gene_or_gene_product
terminal	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
JNK1	B-Gene_or_gene_product
)	O
,	O
and	O
p38	B-Gene_or_gene_product
were	O
activated	O
after	O
denudation	O
,	O
the	O
activation	O
of	O
ERK1	B-Gene_or_gene_product
and	O
p38	B-Gene_or_gene_product
was	O
more	O
rapid	O
and	O
prominent	O
.	O

PD98059	B-Simple_chemical
,	O
a	O
specific	O
MAPK	B-Gene_or_gene_product
/	I-Gene_or_gene_product
ERK	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
(	I-Gene_or_gene_product
MEK	I-Gene_or_gene_product
)	I-Gene_or_gene_product
1	I-Gene_or_gene_product
inhibitor	O
,	O
did	O
not	O
affect	O
the	O
induction	O
of	O
ets	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
mRNA	O
,	O
whereas	O
SB203580	B-Simple_chemical
,	O
a	O
specific	O
p38	B-Gene_or_gene_product
inhibitor	O
,	O
almost	O
completely	O
abrogated	O
its	O
induction	O
.	O

These	O
results	O
indicate	O
that	O
Ets	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
is	O
induced	O
in	O
ECs	B-Cell
after	O
denudation	O
through	O
activation	O
of	O
p38	B-Gene_or_gene_product
.	O

This	O
induction	O
of	O
Ets	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
may	O
be	O
relevant	O
for	O
reendothelialization	O
by	O
regulating	O
the	O
expression	O
of	O
certain	O
genes	O
.	O

SV40	B-Organism
Tag	B-Gene_or_gene_product
transformation	O
of	O
the	O
normal	O
invasive	O
trophoblast	B-Cell
results	O
in	O
a	O
premalignant	O
phenotype	O
.	O

I	O
.	O

Mechanisms	O
responsible	O
for	O
hyperinvasiveness	O
and	O
resistance	O
to	O
anti	O
-	O
invasive	O
action	O
of	O
TGFbeta	B-Gene_or_gene_product
.	O

Invasion	O
of	O
the	O
uterus	B-Organ
by	O
first	O
trimester	O
human	B-Organism
placental	B-Cell
extravillous	I-Cell
trophoblast	I-Cell
(	I-Cell
EVT	I-Cell
)	I-Cell
cells	I-Cell
depends	O
on	O
mechanisms	O
shared	O
by	O
malignant	B-Cell
cells	I-Cell
.	O

However	O
,	O
unlike	O
tumor	B-Cancer
invasion	O
,	O
trophoblast	B-Cell
invasion	O
of	O
the	O
uterus	B-Organ
is	O
stringently	O
controlled	O
in	O
situ	O
by	O
local	O
molecules	O
such	O
as	O
transforming	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	I-Gene_or_gene_product
TGF	I-Gene_or_gene_product
)	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
.	O

Since	O
EVT	B-Cell
cells	I-Cell
possess	O
active	O
invasion	O
-	O
associated	O
genes	O
but	O
are	O
nontumorigenic	O
,	O
our	O
objective	O
was	O
to	O
induce	O
premalignant	O
and	O
then	O
malignant	O
phenotype	O
into	O
a	O
normal	O
EVT	B-Cell
cell	I-Cell
line	I-Cell
in	O
order	O
to	O
identify	O
the	O
molecular	O
basis	O
of	O
tumor	B-Cancer
progression	O
.	O

Simian	B-Organism
virus	I-Organism
40	I-Organism
large	B-Gene_or_gene_product
T	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
(	O
SV40	B-Organism
Tag	B-Gene_or_gene_product
)	O
was	O
introduced	O
into	O
a	O
normal	O
human	B-Organism
first	O
trimester	O
invasive	O
EVT	B-Cell
cell	I-Cell
line	I-Cell
,	O
HTR8	B-Cell
,	O
established	O
in	O
our	O
laboratory	O
.	O

Since	O
the	O
HTR8	B-Cell
line	I-Cell
has	O
a	O
limited	O
in	O
vitro	O
lifespan	O
of	O
12	O
-	O
15	O
passages	O
,	O
SV40	B-Organism
Tag	B-Gene_or_gene_product
-	O
transformed	O
cells	B-Cell
were	O
selected	O
on	O
the	O
basis	O
of	O
extended	O
lifespan	O
.	O

A	O
long	O
-	O
lived	O
line	B-Cell
,	O
RSVT	B-Cell
-	I-Cell
2	I-Cell
,	O
was	O
produced	O
and	O
an	O
immortalized	O
subclone	O
,	O
RSVT2	B-Cell
/	I-Cell
C	I-Cell
,	O
was	O
further	O
derived	O
under	O
a	O
forced	O
crisis	O
regimen	O
.	O

We	O
examined	O
transformation	O
-	O
induced	O
alterations	O
in	O
proliferative	O
and	O
invasive	O
abilities	O
,	O
responses	O
to	O
the	O
invasion	O
and	O
proliferation	O
-	O
regulating	O
growth	O
factor	O
TGFbeta	B-Gene_or_gene_product
and	O
changes	O
in	O
gene	O
expression	O
for	O
invasion	O
-	O
associated	O
enzymes	O
or	O
enzyme	O
inhibitors	O
.	O

RSVT	B-Cell
-	I-Cell
2	I-Cell
and	O
RSVT2	B-Cell
/	I-Cell
C	I-Cell
cell	I-Cell
lines	I-Cell
were	O
hyperproliferative	O
and	O
hyperinvasive	O
when	O
compared	O
with	O
the	O
parental	O
HTR8	B-Cell
cell	I-Cell
line	I-Cell
.	O

They	O
were	O
also	O
variably	O
resistant	O
to	O
the	O
anti	O
-	O
proliferative	O
and	O
anti	O
-	O
invasive	O
signals	O
from	O
TGFbeta	B-Gene_or_gene_product
.	O

Since	O
both	O
cell	B-Cell
lines	I-Cell
remained	O
non	O
-	O
tumorigenic	O
in	O
nude	B-Organism
mice	I-Organism
,	O
these	O
properties	O
indicate	O
that	O
they	O
attained	O
a	O
premalignant	O
phenotype	O
.	O

Both	O
cell	B-Cell
lines	I-Cell
showed	O
reduced	O
expression	O
of	O
tissue	B-Gene_or_gene_product
inhibitor	I-Gene_or_gene_product
of	I-Gene_or_gene_product
metalloproteases	I-Gene_or_gene_product
(	I-Gene_or_gene_product
TIMP	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
while	O
TIMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
plasminogen	B-Gene_or_gene_product
activator	I-Gene_or_gene_product
inhibitor	I-Gene_or_gene_product
(	I-Gene_or_gene_product
PAI	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	I-Gene_or_gene_product
I	I-Gene_or_gene_product
expression	O
was	O
was	O
also	O
reduced	O
in	O
RSVT2	B-Cell
/	I-Cell
C	I-Cell
cells	I-Cell
,	O
thus	O
contributing	O
to	O
their	O
hyperinvasiveness	O
.	O

Their	O
resistance	O
to	O
the	O
anti	O
-	O
invasive	O
action	O
of	O
TGFbeta	B-Gene_or_gene_product
was	O
explained	O
by	O
the	O
failure	O
of	O
TGFbeta	B-Gene_or_gene_product
to	O
upregulate	O
TIMPs	B-Gene_or_gene_product
and	O
PAI	B-Gene_or_gene_product
-	I-Gene_or_gene_product
I	I-Gene_or_gene_product
,	O
in	O
contrast	O
to	O
the	O
TGFbeta	B-Gene_or_gene_product
-	O
induced	O
upregulation	O
noted	O
in	O
parental	O
HTR8	B-Cell
cells	I-Cell
.	O

Expression	O
of	O
two	O
type	B-Gene_or_gene_product
II	I-Gene_or_gene_product
-	I-Gene_or_gene_product
like	I-Gene_or_gene_product
tumor	I-Gene_or_gene_product
hexokinase	I-Gene_or_gene_product
RNA	O
transcripts	O
in	O
cancer	B-Cell
cell	I-Cell
lines	I-Cell
.	O

To	O
maintain	O
an	O
elevated	O
glycolytic	O
rate	O
,	O
cancerous	O
or	O
proliferating	O
cells	B-Cell
alter	O
the	O
expression	O
pattern	O
of	O
rate	O
limiting	O
glycolytic	B-Gene_or_gene_product
enzymes	I-Gene_or_gene_product
.	O

Since	O
glucose	B-Simple_chemical
phosphorylation	O
is	O
the	O
first	O
step	O
in	O
glycolysis	O
,	O
hexokinase	B-Gene_or_gene_product
(	O
HK	B-Gene_or_gene_product
)	O
,	O
the	O
first	O
rate	O
limiting	O
glycolytic	B-Gene_or_gene_product
enzyme	I-Gene_or_gene_product
,	O
can	O
play	O
a	O
key	O
regulatory	O
role	O
in	O
this	O
process	O
.	O

A	O
low	O
-	O
Km	O
,	O
mitochondrial	B-Cellular_component
type	B-Gene_or_gene_product
II	I-Gene_or_gene_product
-	I-Gene_or_gene_product
like	I-Gene_or_gene_product
tumor	I-Gene_or_gene_product
HK	I-Gene_or_gene_product
is	O
described	O
as	O
the	O
predominant	O
form	O
in	O
hepatomas	B-Cancer
.	O

However	O
,	O
recent	O
identification	O
of	O
a	O
high	O
-	O
Km	O
glucose	B-Simple_chemical
phosphorylating	O
activity	O
in	O
a	O
range	O
of	O
cancer	B-Cell
cells	I-Cell
prompted	O
us	O
to	O
characterize	O
glucose	B-Simple_chemical
phosphorylating	O
enzymes	O
of	O
cancer	B-Cell
cells	I-Cell
at	O
the	O
molecular	O
level	O
.	O

Highly	O
sensitive	O
reverse	O
-	O
transcription	O
polymerase	O
chain	O
reaction	O
identifies	O
an	O
induction	O
and	O
overexpression	O
of	O
a	O
type	B-Gene_or_gene_product
II	I-Gene_or_gene_product
-	I-Gene_or_gene_product
like	I-Gene_or_gene_product
tumor	I-Gene_or_gene_product
HK	I-Gene_or_gene_product
RNA	O
in	O
a	O
range	O
of	O
cancer	B-Cell
cell	I-Cell
lines	I-Cell
irrespective	O
of	O
tissue	B-Tissue
origin	O
.	O

In	O
addition	O
,	O
we	O
report	O
here	O
the	O
identification	O
of	O
two	O
RNA	O
transcripts	O
of	O
type	B-Gene_or_gene_product
II	I-Gene_or_gene_product
-	I-Gene_or_gene_product
like	I-Gene_or_gene_product
tumor	I-Gene_or_gene_product
HK	I-Gene_or_gene_product
of	O
approximately	O
5	O
.	O
5	O
and	O
approximately	O
4	O
.	O
0	O
kb	O
in	O
these	O
cancer	B-Cell
cells	I-Cell
lines	I-Cell
,	O
including	O
muscle	B-Organ
-	O
derived	O
L6	B-Cell
myoblast	I-Cell
cells	I-Cell
.	O

Interestingly	O
,	O
under	O
normal	O
conditions	O
muscle	B-Cell
cells	I-Cell
express	O
only	O
a	O
approximately	O
5	O
.	O
5	O
-	O
kb	O
type	B-Gene_or_gene_product
II	I-Gene_or_gene_product
HK	I-Gene_or_gene_product
RNA	O
transcript	O
.	O

A	O
significant	O
amount	O
of	O
type	B-Gene_or_gene_product
I	I-Gene_or_gene_product
HK	I-Gene_or_gene_product
RNA	O
was	O
also	O
found	O
expressed	O
in	O
cancer	B-Cell
cell	I-Cell
lines	I-Cell
.	O

RNA	O
encoding	O
glucokinase	B-Gene_or_gene_product
(	O
GK	B-Gene_or_gene_product
)	O
,	O
the	O
high	O
-	O
Km	O
HK	B-Gene_or_gene_product
isozyme	O
,	O
was	O
found	O
only	O
in	O
cancer	B-Cell
cells	I-Cell
originating	O
from	O
liver	B-Organ
and	O
pancreas	B-Organ
,	O
which	O
express	O
GK	B-Gene_or_gene_product
under	O
normal	O
conditions	O
.	O

Homeobox	B-Gene_or_gene_product
genes	O
in	O
hematopoiesis	O
and	O
leukemogenesis	O
.	O

Homeobox	B-Gene_or_gene_product
genes	O
have	O
a	O
strikingly	O
conserved	O
61	O
-	O
amino	B-Amino_acid
acid	I-Amino_acid
sequence	O
,	O
encoding	O
DNA	B-Cellular_component
-	O
binding	O
domain	O
.	O

Some	O
homeobox	B-Gene_or_gene_product
genes	O
,	O
located	O
in	O
four	O
clusters	O
are	O
designated	O
HOX	B-Gene_or_gene_product
A	I-Gene_or_gene_product
through	O
D	B-Gene_or_gene_product
,	O
while	O
others	O
are	O
known	O
as	O
divergent	O
homeobox	B-Gene_or_gene_product
genes	O
.	O

Many	O
researchers	O
have	O
demonstrated	O
that	O
hematopoietic	B-Cell
cells	I-Cell
express	O
many	O
homeobox	B-Gene_or_gene_product
genes	O
.	O

Induced	O
over	O
-	O
and	O
under	O
-	O
expression	O
of	O
these	O
genes	O
has	O
been	O
used	O
to	O
demonstrate	O
their	O
affect	O
on	O
some	O
aspects	O
of	O
hematopoiesis	O
and	O
leukemogenesis	O
.	O

Recently	O
,	O
knock	O
-	O
out	O
mice	B-Organism
of	O
homeobox	B-Gene_or_gene_product
genes	O
by	O
targeted	O
disruption	O
have	O
also	O
been	O
used	O
to	O
examine	O
their	O
hematological	O
effect	O
.	O

There	O
are	O
examples	O
of	O
the	O
aberrant	O
expression	O
of	O
a	O
homeobox	B-Gene_or_gene_product
gene	O
causing	O
leukemias	B-Cancer
in	O
humans	B-Organism
.	O

In	O
pre	B-Cancer
-	I-Cancer
B	I-Cancer
acute	I-Cancer
lymphoblastic	I-Cancer
leukemia	I-Cancer
(	O
ALL	B-Cancer
)	O
with	O
t	O
(	O
1	O
;	O
19	O
)	O
translocation	O
,	O
a	O
fusion	O
protein	O
is	O
created	O
between	O
E2A	B-Gene_or_gene_product
and	O
a	O
homeobox	B-Gene_or_gene_product
gene	O
PBX	B-Gene_or_gene_product
.	O

In	O
T	B-Cancer
-	I-Cancer
cell	I-Cancer
ALL	I-Cancer
with	O
t	O
(	O
10	O
;	O
14	O
)	O
translocation	O
,	O
the	O
HOX	B-Gene_or_gene_product
11	I-Gene_or_gene_product
gene	O
is	O
deregulated	O
.	O

In	O
acute	B-Cancer
myeloid	I-Cancer
leukemia	I-Cancer
(	O
AML	B-Cancer
)	O
with	O
t	O
(	O
7	O
;	O
11	O
)	O
translocation	O
,	O
the	O
HOX	B-Gene_or_gene_product
A9	I-Gene_or_gene_product
gene	O
is	O
rearranged	O
.	O

In	O
this	O
review	O
article	O
,	O
many	O
functions	O
of	O
homeobox	B-Gene_or_gene_product
genes	O
both	O
at	O
the	O
early	O
stem	B-Cell
cell	I-Cell
level	O
as	O
well	O
as	O
at	O
the	O
later	O
stages	O
of	O
hematopoietic	O
differentiation	O
,	O
and	O
the	O
leukemogenic	O
effect	O
of	O
altered	O
homeobox	B-Gene_or_gene_product
genes	O
are	O
discussed	O
.	O

Store	O
-	O
operated	O
calcium	B-Simple_chemical
entry	O
promotes	O
shape	O
change	O
in	O
pulmonary	B-Cell
endothelial	I-Cell
cells	I-Cell
expressing	O
Trp1	B-Gene_or_gene_product
.	O

Activation	O
of	O
Ca2	B-Simple_chemical
+	I-Simple_chemical
entry	O
is	O
known	O
to	O
produce	O
endothelial	B-Cell
cell	I-Cell
shape	O
change	O
,	O
leading	O
to	O
increased	O
permeability	O
,	O
leukocyte	B-Cell
migration	O
,	O
and	O
initiation	O
of	O
angiogenesis	O
in	O
conduit	B-Cell
-	I-Cell
vessel	I-Cell
endothelial	I-Cell
cells	I-Cell
.	O

The	O
mode	O
of	O
Ca2	B-Simple_chemical
+	I-Simple_chemical
entry	O
regulating	O
cell	B-Cell
shape	O
is	O
unknown	O
.	O

We	O
hypothesized	O
that	O
activation	O
of	O
store	O
-	O
operated	O
Ca2	B-Simple_chemical
+	I-Simple_chemical
channels	O
(	O
SOCs	O
)	O
is	O
sufficient	O
to	O
promote	O
cell	B-Cell
shape	O
change	O
necessary	O
for	O
these	O
processes	O
.	O

SOC	O
activation	O
in	O
rat	B-Organism
pulmonary	B-Cell
arterial	I-Cell
endothelial	I-Cell
cells	I-Cell
increased	O
free	O
cytosolic	B-Organism_substance
Ca2	B-Simple_chemical
+	I-Simple_chemical
that	O
was	O
dependent	O
on	O
a	O
membrane	B-Cellular_component
current	O
having	O
a	O
net	O
inward	O
component	O
of	O
5	O
.	O
45	O
+	O
/	O
-	O
0	O
.	O
90	O
pA	O
/	O
pF	O
at	O
-	O
80	O
mV	O
.	O

Changes	O
in	O
endothelial	B-Cell
cell	I-Cell
shape	O
accompanied	O
SOC	O
activation	O
and	O
were	O
dependent	O
on	O
Ca2	B-Simple_chemical
+	I-Simple_chemical
entry	O
-	O
induced	O
reconfiguration	O
of	O
peripheral	O
(	O
cortical	B-Multi-tissue_structure
)	O
filamentous	B-Gene_or_gene_product
actin	I-Gene_or_gene_product
(	O
F	B-Gene_or_gene_product
-	I-Gene_or_gene_product
actin	I-Gene_or_gene_product
)	O
.	O

Because	O
the	O
identity	O
of	O
pulmonary	O
endothelial	B-Cell
SOCs	O
is	O
unknown	O
,	O
but	O
mammalian	O
homologues	O
of	O
the	O
Drosophila	B-Organism
melanogaster	I-Organism
transient	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
potential	I-Gene_or_gene_product
(	O
trp	B-Gene_or_gene_product
)	O
gene	O
have	O
been	O
proposed	O
to	O
form	O
Ca2	B-Simple_chemical
+	I-Simple_chemical
entry	O
channels	O
in	O
nonexcitable	O
cells	B-Cell
,	O
we	O
performed	O
RT	O
-	O
PCR	O
using	O
Trp	B-Gene_or_gene_product
oligonucleotide	O
primers	O
in	O
both	O
rat	B-Organism
and	O
human	B-Organism
pulmonary	B-Cell
arterial	I-Cell
endothelial	I-Cell
cells	I-Cell
.	O

Both	O
cell	B-Cell
types	O
were	O
found	O
to	O
express	O
Trp1	B-Gene_or_gene_product
,	O
but	O
neither	O
expressed	O
Trp3	B-Gene_or_gene_product
nor	O
Trp6	B-Gene_or_gene_product
.	O

Our	O
study	O
indicates	O
that	O
1	O
)	O
Ca2	B-Simple_chemical
+	I-Simple_chemical
entry	O
in	O
pulmonary	B-Cell
endothelial	I-Cell
cells	I-Cell
through	O
SOCs	O
produces	O
cell	B-Cell
shape	O
change	O
that	O
is	O
dependent	O
on	O
site	O
-	O
specific	O
rearrangement	O
of	O
the	O
microfilamentous	B-Cellular_component
cytoskeleton	I-Cellular_component
and	O
2	O
)	O
Trp1	B-Gene_or_gene_product
may	O
be	O
a	O
component	O
of	O
pulmonary	B-Cell
endothelial	I-Cell
SOCs	O
.	O

Glucose	B-Simple_chemical
and	O
lactate	B-Simple_chemical
metabolism	O
in	O
C6	B-Cell
glioma	I-Cell
cells	I-Cell
:	O
evidence	O
for	O
the	O
preferential	O
utilization	O
of	O
lactate	B-Simple_chemical
for	O
cell	B-Cell
oxidative	O
metabolism	O
.	O

13C	B-Simple_chemical
and	O
1H	B-Simple_chemical
nuclear	O
magnetic	O
resonance	O
spectroscopy	O
(	O
NMR	O
)	O
was	O
used	O
to	O
investigate	O
the	O
metabolism	O
of	O
L	B-Simple_chemical
-	I-Simple_chemical
lactate	I-Simple_chemical
and	O
D	B-Simple_chemical
-	I-Simple_chemical
glucose	I-Simple_chemical
in	O
C6	B-Cell
glioma	I-Cell
cells	I-Cell
.	O

The	O
13C	B-Simple_chemical
enrichment	O
of	O
cell	B-Cell
metabolites	O
was	O
examined	O
after	O
a	O
4	O
-	O
h	O
incubation	O
in	O
media	O
containing	O
5	O
.	O
5	O
mM	O
glucose	B-Simple_chemical
and	O
11	O
mM	O
lactate	B-Simple_chemical
,	O
each	O
metabolite	O
being	O
alternatively	O
labelled	O
with	O
either	O
[	B-Simple_chemical
1	I-Simple_chemical
-	I-Simple_chemical
13C	I-Simple_chemical
]	I-Simple_chemical
D	I-Simple_chemical
-	I-Simple_chemical
glucose	I-Simple_chemical
or	O
[	B-Simple_chemical
3	I-Simple_chemical
-	I-Simple_chemical
13C	I-Simple_chemical
]	I-Simple_chemical
L	I-Simple_chemical
-	I-Simple_chemical
lactate	I-Simple_chemical
.	O

The	O
results	O
indicated	O
that	O
exogenous	O
lactate	B-Simple_chemical
was	O
the	O
major	O
substrate	O
for	O
oxidative	O
metabolism	O
.	O

They	O
were	O
consistent	O
with	O
the	O
concept	O
of	O
the	O
existence	O
of	O
2	O
pools	O
of	O
both	O
lactate	B-Simple_chemical
and	O
pyruvate	B-Simple_chemical
,	O
of	O
which	O
1	O
pool	O
was	O
closely	O
connected	O
with	O
exogenous	O
lactate	B-Simple_chemical
and	O
oxidative	O
metabolism	O
,	O
and	O
the	O
other	O
pool	O
was	O
closely	O
related	O
to	O
glycolysis	O
and	O
disconnected	O
from	O
oxidative	O
metabolism	O
.	O

The	O
molecular	O
basis	O
of	O
this	O
behaviour	O
could	O
be	O
related	O
to	O
different	O
locations	O
for	O
the	O
lactate	B-Gene_or_gene_product
dehydrogenase	I-Gene_or_gene_product
isoenzymes	I-Gene_or_gene_product
,	O
as	O
suggested	O
by	O
their	O
immunohistochemical	O
labelling	O
.	O

Tumors	B-Cancer
of	O
the	O
retinal	B-Tissue
pigment	I-Tissue
epithelium	I-Tissue
metastasize	O
to	O
inguinal	O
lymph	B-Multi-tissue_structure
nodes	I-Multi-tissue_structure
and	O
spleen	B-Organ
in	O
tyrosinase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
related	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
1	I-Gene_or_gene_product
/	O
SV40	B-Gene_or_gene_product
T	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
transgenic	O
mice	O
.	O

The	O
pigment	B-Tissue
epithelium	I-Tissue
of	O
the	O
retina	B-Multi-tissue_structure
(	O
RPE	B-Tissue
)	O
is	O
derived	O
from	O
the	O
optic	B-Multi-tissue_structure
cup	I-Multi-tissue_structure
and	O
is	O
essential	O
for	O
function	O
and	O
development	O
of	O
the	O
eye	B-Organ
.	O

We	O
produced	O
a	O
transgenic	O
mouse	B-Organism
line	O
that	O
expresses	O
simian	B-Organism
virus	I-Organism
(	O
SV40	B-Organism
)	O
transforming	O
sequences	O
under	O
control	O
of	O
the	O
1	O
.	O
4	O
kb	O
tyrosinase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
related	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
TRP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
promoter	O
,	O
targeting	O
expression	O
of	O
T	B-Gene_or_gene_product
antigen	I-Gene_or_gene_product
(	O
Tag	B-Gene_or_gene_product
)	O
to	O
the	O
RPE	B-Tissue
.	O

In	O
transgenic	O
embryos	B-Developing_anatomical_structure
,	O
RPE	B-Cell
cells	I-Cell
proliferated	O
in	O
the	O
anterior	B-Multi-tissue_structure
part	I-Multi-tissue_structure
of	O
the	O
eye	B-Organ
and	O
near	O
the	O
optic	B-Multi-tissue_structure
nerve	I-Multi-tissue_structure
.	O

This	O
resulted	O
in	O
formation	O
of	O
tumors	B-Cancer
,	O
which	O
were	O
pigmented	O
and	O
of	O
epithelial	B-Tissue
origin	O
.	O

In	O
3	O
months	O
-	O
old	O
mice	B-Organism
,	O
pigmented	B-Cell
cells	I-Cell
were	O
detected	O
in	O
spleen	B-Organ
and	O
inguinal	B-Multi-tissue_structure
lymph	I-Multi-tissue_structure
nodes	I-Multi-tissue_structure
.	O

In	O
spleen	B-Organ
,	O
tyrosinase	B-Gene_or_gene_product
,	O
TRP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
SV40	B-Organism
Tag	B-Gene_or_gene_product
were	O
expressed	O
and	O
tyrosinase	B-Gene_or_gene_product
was	O
enzymatically	O
active	O
.	O

Pigmented	O
regions	O
were	O
positive	O
for	O
an	O
epithelial	B-Tissue
marker	O
,	O
cytokeratin	B-Gene_or_gene_product
.	O

Cell	B-Cell
lines	I-Cell
were	O
established	O
from	O
tumor	B-Cancer
and	O
metastases	O
and	O
kept	O
in	O
culture	O
for	O
more	O
than	O
2	O
months	O
.	O

These	O
were	O
pigmented	O
,	O
and	O
maintained	O
expression	O
of	O
tyrosinase	B-Gene_or_gene_product
,	O
TRP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
cytokeratin	B-Gene_or_gene_product
and	O
SV40	B-Organism
Tag	B-Gene_or_gene_product
.	O

This	O
demonstrates	O
that	O
RPE	B-Cell
tumor	I-Cell
cells	I-Cell
metastasize	O
to	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
and	O
spleen	B-Organ
.	O

In	O
conclusion	O
,	O
the	O
metastasis	O
from	O
TRP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
/	O
Tag	B-Gene_or_gene_product
RPE	O
tumors	O
towards	O
spleen	B-Organ
and	O
lymph	B-Multi-tissue_structure
nodes	I-Multi-tissue_structure
serves	O
as	O
potential	O
tool	O
to	O
investigate	O
biology	O
and	O
metastasis	O
of	O
tumors	B-Cancer
derived	O
from	O
the	O
pigment	B-Tissue
epithelium	I-Tissue
.	O

Effect	O
of	O
U	B-Gene_or_gene_product
-	I-Gene_or_gene_product
995	I-Gene_or_gene_product
,	O
a	O
potent	O
shark	B-Organism
cartilage	B-Tissue
-	O
derived	O
angiogenesis	O
inhibitor	O
,	O
on	O
anti	O
-	O
angiogenesis	O
and	O
anti	O
-	O
tumor	B-Cancer
activities	O
.	O

BACKGROUND	O
:	O
A	O
potent	O
angiogenesis	O
inhibitor	O
,	O
U	B-Gene_or_gene_product
-	I-Gene_or_gene_product
995	I-Gene_or_gene_product
,	O
has	O
been	O
purified	O
from	O
the	O
cartilage	B-Tissue
of	O
the	O
blue	B-Organism
shark	I-Organism
(	O
Prionace	B-Organism
glauca	I-Organism
)	O
.	O

U	B-Gene_or_gene_product
-	I-Gene_or_gene_product
995	I-Gene_or_gene_product
is	O
composed	O
of	O
two	O
single	O
peptides	O
with	O
molecular	O
mass	O
of	O
10	O
and	O
14	O
kDa	O
,	O
respectively	O
.	O

MATERIALS	O
AND	O
METHODS	O
:	O
U	B-Gene_or_gene_product
-	I-Gene_or_gene_product
995	I-Gene_or_gene_product
was	O
designed	O
to	O
study	O
human	B-Cell
umbilical	I-Cell
vein	I-Cell
endothelial	I-Cell
cell	I-Cell
(	O
HUVEC	B-Cell
)	O
migration	O
and	O
proliferation	O
in	O
vitro	O
and	O
angiogenesis	O
induced	O
by	O
TNF	B-Gene_or_gene_product
alpha	I-Gene_or_gene_product
in	O
chicken	B-Organism
chorioallantoic	B-Multi-tissue_structure
membrane	I-Multi-tissue_structure
(	O
CAM	B-Multi-tissue_structure
)	O
.	O

Furthermore	O
,	O
we	O
determined	O
the	O
ability	O
of	O
U	B-Gene_or_gene_product
-	I-Gene_or_gene_product
995	I-Gene_or_gene_product
to	O
inhibiting	O
tumor	B-Cell
cell	I-Cell
growth	O
and	O
metastasis	O
.	O

RESULTS	O
:	O
U	B-Gene_or_gene_product
-	I-Gene_or_gene_product
995	I-Gene_or_gene_product
(	O
15	O
and	O
30	O
micrograms	O
/	O
ml	O
)	O
markedly	O
inhibited	O
HUVEC	B-Cell
migration	O
and	O
,	O
at	O
15	O
-	O
50	O
micrograms	O
/	O
ml	O
produced	O
a	O
dose	O
-	O
dependent	O
decline	O
in	O
[	O
3H	O
]	O
-	O
thymidine	B-Simple_chemical
incorporation	O
.	O

30	O
and	O
50	O
micrograms	O
/	O
ml	O
of	O
U	B-Gene_or_gene_product
-	I-Gene_or_gene_product
995	I-Gene_or_gene_product
,	O
when	O
added	O
to	O
TNF	B-Gene_or_gene_product
alpha	I-Gene_or_gene_product
-	O
induced	O
angiogenesis	O
caused	O
discontinuous	O
and	O
disrupted	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
.	O

Moreover	O
,	O
U	B-Gene_or_gene_product
-	I-Gene_or_gene_product
995	I-Gene_or_gene_product
(	O
30	O
micrograms	O
/	O
ml	O
)	O
markedly	O
prevented	O
collagenase	B-Gene_or_gene_product
-	O
induced	O
collagenolysis	O
.	O

In	O
addition	O
,	O
when	O
200	O
micrograms	O
U	B-Gene_or_gene_product
-	I-Gene_or_gene_product
995	I-Gene_or_gene_product
was	O
injected	O
i	O
.	O
p	O
.	O
into	O
mice	B-Organism
it	O
suppressed	O
sarcoma	B-Cell
-	I-Cell
180	I-Cell
cell	I-Cell
growth	O
and	O
B16	B-Cell
-	I-Cell
F10	I-Cell
mouse	I-Cell
melanoma	I-Cell
cell	I-Cell
metastasis	O
in	O
vivo	O
.	O

CONCLUSIONS	O
:	O
These	O
results	O
suggest	O
that	O
the	O
anti	O
-	O
angiogenic	O
effects	O
of	O
U	B-Gene_or_gene_product
-	I-Gene_or_gene_product
995	I-Gene_or_gene_product
may	O
be	O
be	O
due	O
to	O
interference	O
with	O
the	O
proliferation	O
and	O
migration	O
of	O
HUVECs	B-Cell
as	O
well	O
as	O
inhibition	O
of	O
collagenolysis	O
,	O
thereby	O
leading	O
to	O
inhibition	O
of	O
both	O
angiogenesis	O
and	O
tumor	B-Cell
cell	I-Cell
growth	O
.	O

