UFT	B-Simple_chemical
and	O
its	O
metabolites	O
inhibit	O
the	O
angiogenesis	O
induced	O
by	O
murine	B-Organism
renal	B-Cancer
cell	I-Cancer
carcinoma	I-Cancer
,	O
as	O
determined	O
by	O
a	O
dorsal	B-Multi-tissue_structure
air	I-Multi-tissue_structure
sac	I-Multi-tissue_structure
assay	O
in	O
mice	B-Organism
.	O

UFT	B-Simple_chemical
,	O
an	O
anticancer	B-Cancer
agent	O
that	O
is	O
composed	O
of	O
tegafur	B-Simple_chemical
(	O
FT	B-Simple_chemical
)	O
and	O
uracil	B-Simple_chemical
at	O
a	O
molar	O
ratio	O
of	O
1	O
:	O
4	O
,	O
is	O
widely	O
used	O
in	O
clinical	O
practice	O
in	O
Japan	O
to	O
treat	O
cancer	B-Cancer
patients	B-Organism
requiring	O
a	O
long	O
-	O
term	O
chemotherapy	O
,	O
and	O
it	O
is	O
associated	O
with	O
few	O
side	O
effects	O
,	O
if	O
any	O
.	O

In	O
this	O
study	O
,	O
we	O
have	O
evaluated	O
the	O
inhibitory	O
effect	O
of	O
UFT	B-Simple_chemical
against	O
RENCA	B-Cell
cell	I-Cell
-	O
induced	O
angiogenesis	O
by	O
a	O
dorsal	O
air	O
sac	O
assay	O
.	O

Marked	O
angiogenesis	O
is	O
induced	O
by	O
implantation	O
of	O
a	O
chamber	O
containing	O
RENCA	B-Cell
cells	I-Cell
into	O
mice	B-Organism
.	O

In	O
this	O
model	O
,	O
UFT	B-Simple_chemical
showed	O
a	O
strong	O
angiogenesis	O
-	O
inhibitory	O
effect	O
,	O
whereas	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
and	O
doxifluridine	B-Simple_chemical
were	O
less	O
effective	O
.	O

Additional	O
experiments	O
revealed	O
FT	B-Simple_chemical
to	O
be	O
effective	O
component	O
of	O
UFT	B-Simple_chemical
;	O
uracil	B-Simple_chemical
remained	O
ineffective	O
in	O
the	O
inhibition	O
of	O
angiogenesis	O
.	O

Moreover	O
,	O
we	O
have	O
found	O
that	O
gamma	B-Simple_chemical
-	I-Simple_chemical
hydroxybutyric	I-Simple_chemical
acid	I-Simple_chemical
and	O
gamma	B-Simple_chemical
-	I-Simple_chemical
butyrolactone	I-Simple_chemical
,	O
the	O
metabolites	O
of	O
FT	B-Simple_chemical
,	O
possess	O
a	O
potent	O
angiogenesis	O
inhibitory	O
effect	O
that	O
is	O
amplified	O
when	O
the	O
compounds	O
are	O
administered	O
by	O
a	O
continuous	O
infusion	O
.	O

This	O
may	O
reflect	O
a	O
transition	O
in	O
blood	B-Organism_substance
concentration	O
of	O
each	O
metabolite	O
resulting	O
from	O
the	O
administration	O
of	O
UFT	B-Simple_chemical
.	O

Similar	O
results	O
were	O
also	O
obtained	O
with	O
respect	O
to	O
5	B-Simple_chemical
-	I-Simple_chemical
FU	I-Simple_chemical
.	O

It	O
was	O
suggested	O
that	O
UFT	B-Simple_chemical
has	O
a	O
stronger	O
angiogenesis	O
-	O
inhibitory	O
effect	O
than	O
did	O
other	O
fluorinated	B-Simple_chemical
pyrimidines	I-Simple_chemical
,	O
partly	O
due	O
to	O
its	O
pharmacokinetic	O
properties	O
characterized	O
by	O
maintaining	O
of	O
higher	O
and	O
long	O
-	O
lasting	O
blood	B-Organism_substance
levels	O
of	O
5	B-Simple_chemical
-	I-Simple_chemical
FU	I-Simple_chemical
and	O
partly	O
due	O
the	O
inhibitory	O
effects	O
derived	O
from	O
gamma	B-Simple_chemical
-	I-Simple_chemical
hydroxybutyric	I-Simple_chemical
acid	I-Simple_chemical
and	O
gamma	B-Simple_chemical
-	I-Simple_chemical
butyrolactone	I-Simple_chemical
,	O
UFT	B-Simple_chemical
-	O
specific	O
metabolites	O
.	O

Nitric	B-Gene_or_gene_product
oxide	I-Gene_or_gene_product
synthase	I-Gene_or_gene_product
inhibition	O
results	O
in	O
synergistic	O
anti	O
-	O
tumour	B-Cancer
activity	O
with	O
melphalan	B-Simple_chemical
and	O
tumour	B-Gene_or_gene_product
necrosis	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
-	O
based	O
isolated	O
limb	B-Organism_subdivision
perfusions	O
.	O

Nitric	B-Simple_chemical
oxide	I-Simple_chemical
(	O
NO	B-Simple_chemical
)	O
is	O
an	O
important	O
molecule	O
in	O
regulating	O
tumour	B-Cancer
blood	B-Organism_substance
flow	O
and	O
stimulating	O
tumour	B-Cancer
angiogenesis	O
.	O

Inhibition	O
of	O
NO	B-Gene_or_gene_product
synthase	I-Gene_or_gene_product
by	O
L	B-Simple_chemical
-	I-Simple_chemical
NAME	I-Simple_chemical
might	O
induce	O
an	O
anti	O
-	O
tumour	B-Cancer
effect	O
by	O
limiting	O
nutrients	O
and	O
oxygen	B-Simple_chemical
to	O
reach	O
tumour	B-Tissue
tissue	I-Tissue
or	O
affecting	O
vascular	B-Multi-tissue_structure
growth	O
.	O

The	O
anti	O
-	O
tumour	B-Cancer
effect	O
of	O
L	B-Simple_chemical
-	I-Simple_chemical
NAME	I-Simple_chemical
after	O
systemic	O
administration	O
was	O
studied	O
in	O
a	O
renal	B-Cancer
subcapsular	I-Cancer
CC531	I-Cancer
adenocarcinoma	I-Cancer
model	O
in	O
rats	B-Organism
.	O

Moreover	O
,	O
regional	O
administration	O
of	O
L	B-Simple_chemical
-	I-Simple_chemical
NAME	I-Simple_chemical
,	O
in	O
combination	O
with	O
TNF	B-Gene_or_gene_product
and	O
melphalan	B-Simple_chemical
,	O
was	O
studied	O
in	O
an	O
isolated	O
limb	B-Organism_subdivision
perfusion	O
(	O
ILP	O
)	O
model	O
using	O
BN175	B-Cancer
soft	I-Cancer
-	I-Cancer
tissue	I-Cancer
sarcomas	I-Cancer
.	O

Systemic	O
treatment	O
with	O
L	B-Simple_chemical
-	I-Simple_chemical
NAME	I-Simple_chemical
inhibited	O
growth	O
of	O
adenocarcinoma	B-Cancer
significantly	O
but	O
was	O
accompanied	O
by	O
impaired	O
renal	B-Organ
function	O
.	O

In	O
ILP	O
,	O
reduced	O
tumour	B-Cancer
growth	O
was	O
observed	O
when	O
L	B-Simple_chemical
-	I-Simple_chemical
NAME	I-Simple_chemical
was	O
used	O
alone	O
.	O

In	O
combination	O
with	O
TNF	B-Gene_or_gene_product
or	O
melphalan	B-Simple_chemical
,	O
L	B-Simple_chemical
-	I-Simple_chemical
NAME	I-Simple_chemical
increased	O
response	O
rates	O
significantly	O
compared	O
to	O
perfusions	O
without	O
L	B-Simple_chemical
-	I-Simple_chemical
NAME	I-Simple_chemical
(	O
0	O
-	O
64	O
%	O
and	O
0	O
-	O
63	O
%	O
respectively	O
)	O
.	O

An	O
additional	O
anti	O
-	O
tumour	B-Cancer
effect	O
was	O
demonstrated	O
when	O
L	B-Simple_chemical
-	I-Simple_chemical
NAME	I-Simple_chemical
was	O
added	O
to	O
the	O
synergistic	O
combination	O
of	O
melphalan	B-Simple_chemical
and	O
TNF	B-Gene_or_gene_product
(	O
responses	O
increased	O
from	O
70	O
to	O
100	O
%	O
)	O
.	O

Inhibition	O
of	O
NO	B-Gene_or_gene_product
synthase	I-Gene_or_gene_product
reduces	O
tumour	B-Cancer
growth	O
both	O
after	O
systemic	O
and	O
regional	O
(	O
ILP	O
)	O
treatment	O
.	O

A	O
synergistic	O
anti	O
-	O
tumour	B-Cancer
effect	O
of	O
L	B-Simple_chemical
-	I-Simple_chemical
NAME	I-Simple_chemical
is	O
observed	O
in	O
combination	O
with	O
melphalan	B-Simple_chemical
and	O
/	O
or	O
TNF	B-Gene_or_gene_product
using	O
ILP	O
.	O

These	O
results	O
indicate	O
a	O
possible	O
role	O
of	O
L	B-Simple_chemical
-	I-Simple_chemical
NAME	I-Simple_chemical
for	O
the	O
treatment	O
of	O
solid	O
tumours	B-Cancer
in	O
a	O
systemic	O
or	O
regional	O
setting	O
.	O

Apoptosis	O
induced	O
by	O
1	B-Simple_chemical
'	I-Simple_chemical
-	I-Simple_chemical
acetoxychavicol	I-Simple_chemical
acetate	I-Simple_chemical
in	O
Ehrlich	B-Cell
ascites	I-Cell
tumor	I-Cell
cells	I-Cell
is	O
associated	O
with	O
modulation	O
of	O
polyamine	O
metabolism	O
and	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
activation	O
.	O

The	O
efficacy	O
of	O
the	O
antitumor	B-Cancer
activity	O
of	O
1	B-Simple_chemical
'	I-Simple_chemical
-	I-Simple_chemical
acetoxychavicol	I-Simple_chemical
acetate	I-Simple_chemical
(	O
ACA	B-Simple_chemical
)	O
,	O
reported	O
to	O
be	O
a	O
suppressor	O
of	O
chemically	O
induced	O
carcinogenesis	O
,	O
was	O
evaluated	O
in	O
Ehrlich	B-Cell
ascites	I-Cell
tumor	I-Cell
cells	I-Cell
.	O

ACA	B-Simple_chemical
treatment	O
resulted	O
in	O
changes	O
in	O
morphology	O
and	O
a	O
dose	O
-	O
dependent	O
suppression	O
of	O
cell	B-Cell
viability	O
.	O

Apoptosis	O
,	O
characterized	O
by	O
nuclear	B-Cellular_component
condensation	O
,	O
membrane	B-Cellular_component
blebbing	O
,	O
cell	B-Cell
shrinkage	O
and	O
a	O
significant	O
induction	O
of	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
-	I-Gene_or_gene_product
like	I-Gene_or_gene_product
protease	I-Gene_or_gene_product
activity	O
at	O
8	O
h	O
in	O
a	O
time	O
-	O
course	O
study	O
were	O
observed	O
.	O

Formation	O
of	O
apoptotic	B-Cellular_component
bodies	I-Cellular_component
was	O
preceded	O
by	O
lowering	O
of	O
intracellular	B-Immaterial_anatomical_entity
polyamines	O
,	O
particularly	O
putrescine	B-Simple_chemical
,	O
and	O
both	O
dose	O
-	O
and	O
time	O
-	O
dependent	O
inhibitory	O
and	O
activation	O
effect	O
by	O
ACA	B-Simple_chemical
on	O
ornithine	B-Gene_or_gene_product
decarboxylase	I-Gene_or_gene_product
(	O
ODC	B-Gene_or_gene_product
)	O
and	O
spermidine	B-Gene_or_gene_product
/	I-Gene_or_gene_product
spermine	I-Gene_or_gene_product
N	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
acetyltransferase	I-Gene_or_gene_product
(	O
SSAT	B-Gene_or_gene_product
)	O
,	O
respectively	O
.	O

Administration	O
of	O
exogenous	O
polyamines	O
prevented	O
ACA	B-Simple_chemical
-	O
induced	O
apoptosis	O
represented	O
by	O
a	O
reduction	O
in	O
the	O
number	O
of	O
apoptotic	B-Cellular_component
bodies	I-Cellular_component
and	O
also	O
caused	O
reduction	O
in	O
the	O
induced	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
-	I-Gene_or_gene_product
like	I-Gene_or_gene_product
protease	I-Gene_or_gene_product
activity	O
at	O
8	O
h	O
.	O

These	O
findings	O
suggest	O
that	O
the	O
anticarcinogenic	O
effects	O
of	O
ACA	B-Simple_chemical
might	O
be	O
partly	O
due	O
to	O
perturbation	O
of	O
the	O
polyamine	O
metabolic	O
pathway	O
and	O
triggering	O
of	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
-	I-Gene_or_gene_product
like	I-Gene_or_gene_product
activity	O
,	O
which	O
result	O
in	O
apoptosis	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
-	I-Gene_or_gene_product
B	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
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
expression	O
in	O
renal	B-Cancer
cell	I-Cancer
carcinomas	I-Cancer
:	O
regulation	O
by	O
the	O
von	B-Gene_or_gene_product
Hippel	I-Gene_or_gene_product
-	I-Gene_or_gene_product
Lindau	I-Gene_or_gene_product
gene	O
and	O
hypoxia	O
.	O

Angiogenesis	O
is	O
essential	O
for	O
tumor	B-Cancer
growth	O
and	O
metastasis	O
.	O

It	O
is	O
regulated	O
by	O
numerous	O
angiogenic	O
factors	O
,	O
one	O
of	O
the	O
most	O
important	O
being	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

Recently	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
B	I-Gene_or_gene_product
and	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
,	O
two	O
new	O
VEGF	B-Gene_or_gene_product
family	O
members	O
,	O
have	O
been	O
identified	O
that	O
bind	O
to	O
the	O
tyrosine	O
kinase	O
receptors	O
flt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
VEGFR1	B-Gene_or_gene_product
)	O
,	O
KDR	B-Gene_or_gene_product
(	O
VEGFR2	B-Gene_or_gene_product
)	O
,	O
and	O
flt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
4	I-Gene_or_gene_product
(	O
VEGFR3	B-Gene_or_gene_product
)	O
.	O

Although	O
the	O
importance	O
of	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
A	I-Gene_or_gene_product
has	O
been	O
shown	O
in	O
renal	B-Cancer
carcinomas	I-Cancer
,	O
the	O
contribution	O
of	O
these	O
new	O
ligands	O
in	O
kidney	B-Cancer
tumors	I-Cancer
is	O
not	O
clear	O
.	O

We	O
have	O
,	O
therefore	O
,	O
measured	O
the	O
mRNA	O
level	O
of	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
B	I-Gene_or_gene_product
and	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
together	O
with	O
their	O
receptors	O
by	O
RNase	B-Gene_or_gene_product
protection	O
assay	O
(	O
RPA	O
)	O
in	O
26	O
normal	B-Multi-tissue_structure
kidney	I-Multi-tissue_structure
samples	I-Multi-tissue_structure
and	O
45	O
renal	B-Cancer
cell	I-Cancer
cancers	I-Cancer
.	O

We	O
observed	O
a	O
significant	O
up	O
-	O
regulation	O
of	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
B	I-Gene_or_gene_product
(	O
P	O
=	O
0	O
.	O
002	O
)	O
but	O
not	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
(	O
P	O
=	O
0	O
.	O
3	O
)	O
in	O
neoplastic	B-Organ
kidney	I-Organ
compared	O
with	O
normal	B-Tissue
tissues	I-Tissue
.	O

In	O
addition	O
,	O
although	O
VEGF	B-Gene_or_gene_product
receptors	I-Gene_or_gene_product
were	O
higher	O
in	O
tumors	B-Cancer
than	O
normal	O
kidney	B-Organ
,	O
there	O
was	O
a	O
significant	O
up	O
-	O
regulation	O
of	O
only	O
flt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
P	O
=	O
0	O
.	O
003	O
)	O
but	O
not	O
KDR	B-Gene_or_gene_product
(	O
P	O
=	O
0	O
.	O
12	O
)	O
or	O
flt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
4	I-Gene_or_gene_product
(	O
P	O
=	O
0	O
.	O
09	O
)	O
.	O

There	O
was	O
also	O
a	O
significant	O
correlation	O
between	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
and	O
both	O
of	O
its	O
receptors	O
flt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
4	I-Gene_or_gene_product
(	O
P	O
=	O
0	O
.	O
006	O
)	O
and	O
KDR	B-Gene_or_gene_product
(	O
P	O
=	O
0	O
.	O
03	O
)	O
but	O
no	O
association	O
between	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
B	I-Gene_or_gene_product
and	O
its	O
receptor	O
flt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
P	O
=	O
0	O
.	O
23	O
)	O
.	O

A	O
significant	O
increase	O
was	O
observed	O
in	O
flt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
P	O
less	O
than	O
0	O
.	O
001	O
)	O
,	O
KDR	B-Gene_or_gene_product
(	O
P	O
=	O
0	O
.	O
02	O
)	O
,	O
and	O
flt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
4	I-Gene_or_gene_product
(	O
P	O
=	O
0	O
.	O
01	O
)	O
but	O
not	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
B	I-Gene_or_gene_product
(	O
P	O
=	O
0	O
.	O
82	O
)	O
or	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
(	O
P	O
=	O
0	O
.	O
52	O
)	O
expression	O
in	O
clear	B-Cell
cell	I-Cell
compared	O
with	O
chromophil	B-Cancer
(	I-Cancer
papillary	I-Cancer
)	I-Cancer
carcinomas	I-Cancer
.	O

No	O
significant	O
association	O
was	O
demonstrated	O
between	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
B	I-Gene_or_gene_product
,	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
,	O
flt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
KDR	B-Gene_or_gene_product
,	O
and	O
flt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
4	I-Gene_or_gene_product
with	O
patient	B-Organism
sex	O
,	O
patient	B-Organism
age	O
,	O
or	O
tumor	B-Cancer
size	O
(	O
P	O
greater	O
than	O
0	O
.	O
05	O
)	O
.	O

The	O
effect	O
of	O
von	B-Gene_or_gene_product
Hippel	I-Gene_or_gene_product
-	I-Gene_or_gene_product
Lindau	I-Gene_or_gene_product
(	O
VHL	B-Gene_or_gene_product
)	O
gene	O
and	O
hypoxia	O
on	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
B	I-Gene_or_gene_product
and	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
expression	O
in	O
the	O
renal	B-Cell
carcinoma	I-Cell
cell	I-Cell
line	I-Cell
786	I-Cell
-	I-Cell
0	I-Cell
transfected	O
with	O
wild	O
-	O
type	O
and	O
mutant	O
VHL	B-Gene_or_gene_product
was	O
determined	O
by	O
growing	O
cells	B-Cell
under	O
21	O
%	O
O2	O
-	O
and	O
0	O
.	O
1	O
%	O
O2	O
.	O

In	O
wild	O
-	O
type	O
VHL	B-Gene_or_gene_product
cells	O
,	O
whereas	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
A	I-Gene_or_gene_product
was	O
significantly	O
up	O
-	O
regulated	O
under	O
hypoxic	O
compared	O
with	O
normoxic	O
conditions	O
(	O
P	O
less	O
than	O
0	O
.	O
001	O
)	O
,	O
expression	O
of	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
was	O
reduced	O
(	O
P	O
less	O
than	O
0	O
.	O
002	O
)	O
.	O

Nevertheless	O
,	O
the	O
repression	O
of	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
was	O
lost	O
in	O
mutant	O
VHL	B-Gene_or_gene_product
cell	O
lines	O
under	O
hypoxia	O
.	O

In	O
contrast	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
B	I-Gene_or_gene_product
was	O
not	O
regulated	O
by	O
VHL	B-Gene_or_gene_product
despite	O
clear	O
up	O
-	O
regulation	O
in	O
vivo	O
.	O

These	O
findings	O
strongly	O
support	O
an	O
enhanced	O
role	O
for	O
this	O
pathway	O
in	O
clear	B-Cancer
cell	I-Cancer
carcinomas	I-Cancer
by	O
regulating	O
angiogenesis	O
and	O
/	O
or	O
lymphangiogenesis	O
.	O

The	O
study	O
shows	O
that	O
clear	B-Cancer
cell	I-Cancer
tumors	I-Cancer
are	O
able	O
to	O
up	O
-	O
regulate	O
angiogenic	O
growth	O
factor	O
receptors	O
more	O
efficiently	O
than	O
chromophil	B-Cancer
(	O
papillary	B-Cancer
)	O
,	O
that	O
clear	B-Cancer
cell	I-Cancer
tumors	I-Cancer
can	O
use	O
pathways	O
independent	O
of	O
VHL	B-Gene_or_gene_product
to	O
regulate	O
angiogenesis	O
,	O
and	O
that	O
this	O
combined	O
regulation	O
may	O
account	O
for	O
their	O
more	O
aggressive	O
phenotype	O
,	O
which	O
suggests	O
that	O
targeting	O
VEGFR1	B-Gene_or_gene_product
(	O
flt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
l	I-Gene_or_gene_product
)	O
may	O
be	O
particularly	O
effective	O
in	O
these	O
tumor	B-Cancer
types	O
.	O

Glucose	B-Simple_chemical
catabolism	O
in	O
cancer	B-Cell
cells	I-Cell
:	O
identification	O
and	O
characterization	O
of	O
a	O
marked	O
activation	O
response	O
of	O
the	O
type	B-Gene_or_gene_product
II	I-Gene_or_gene_product
hexokinase	I-Gene_or_gene_product
gene	O
to	O
hypoxic	O
conditions	O
.	O

One	O
of	O
the	O
most	O
common	O
signatures	O
of	O
highly	O
malignant	B-Cancer
tumors	I-Cancer
is	O
their	O
capacity	O
to	O
metabolize	O
more	O
glucose	B-Simple_chemical
to	O
lactic	B-Simple_chemical
acid	I-Simple_chemical
than	O
their	O
tissues	B-Tissue
of	O
origin	O
.	O

Hepatomas	B-Cancer
exhibiting	O
this	O
phenotype	O
are	O
dependent	O
on	O
the	O
high	O
expression	O
of	O
type	B-Gene_or_gene_product
II	I-Gene_or_gene_product
hexokinase	I-Gene_or_gene_product
,	O
which	O
supplies	O
such	O
tumors	B-Cancer
with	O
abundant	O
amounts	O
of	O
glucose	B-Simple_chemical
6	I-Simple_chemical
-	I-Simple_chemical
phosphate	I-Simple_chemical
,	O
a	O
significant	O
carbon	B-Simple_chemical
and	O
energy	O
source	O
especially	O
under	O
hypoxic	O
conditions	O
.	O

Here	O
we	O
report	O
that	O
the	O
distal	O
region	O
of	O
the	O
hepatoma	B-Cancer
type	B-Gene_or_gene_product
II	I-Gene_or_gene_product
hexokinase	I-Gene_or_gene_product
promoter	O
displays	O
consensus	O
motifs	O
for	O
hypoxia	B-Gene_or_gene_product
-	I-Gene_or_gene_product
inducible	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
that	O
overlap	O
E	O
-	O
box	O
sequences	O
known	O
to	O
be	O
related	O
in	O
other	O
gene	O
promoters	O
to	O
glucose	B-Simple_chemical
response	O
.	O

Moreover	O
,	O
we	O
show	O
that	O
subjecting	O
transfected	O
hepatoma	B-Cell
cells	I-Cell
to	O
hypoxic	O
conditions	O
activates	O
the	O
type	B-Gene_or_gene_product
II	I-Gene_or_gene_product
hexokinase	I-Gene_or_gene_product
promoter	O
almost	O
3	O
-	O
fold	O
,	O
a	O
value	O
that	O
approaches	O
7	O
-	O
fold	O
in	O
the	O
presence	O
of	O
glucose	B-Simple_chemical
.	O

Consistent	O
with	O
these	O
findings	O
is	O
the	O
induction	O
under	O
hypoxic	O
conditions	O
of	O
the	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
protein	O
.	O

Reporter	O
gene	O
analyses	O
with	O
a	O
series	O
of	O
nested	O
deletion	O
mutants	O
of	O
the	O
hepatoma	B-Cancer
type	B-Gene_or_gene_product
II	I-Gene_or_gene_product
hexokinase	I-Gene_or_gene_product
promoter	O
show	O
that	O
a	O
significant	O
fraction	O
of	O
the	O
total	O
activation	O
observed	O
under	O
hypoxic	O
conditions	O
localizes	O
to	O
the	O
distal	O
region	O
where	O
the	O
overlapping	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
/	O
E	O
-	O
box	O
sequences	O
are	O
located	O
.	O

Finally	O
,	O
DNase	B-Gene_or_gene_product
I	I-Gene_or_gene_product
footprint	O
analysis	O
with	O
a	O
segment	O
of	O
the	O
promoter	O
containing	O
these	O
elements	O
reveals	O
the	O
binding	O
of	O
several	O
nuclear	B-Cellular_component
proteins	O
.	O

In	O
summary	O
,	O
these	O
novel	O
studies	O
identify	O
and	O
characterize	O
a	O
marked	O
glucose	B-Simple_chemical
-	O
modulated	O
activation	O
response	O
of	O
the	O
type	B-Gene_or_gene_product
II	I-Gene_or_gene_product
hexokinase	I-Gene_or_gene_product
gene	O
to	O
hypoxic	O
conditions	O
within	O
highly	O
glycolytic	O
hepatoma	B-Cell
cells	I-Cell
,	O
a	O
property	O
that	O
may	O
help	O
assure	O
that	O
such	O
cells	B-Cell
exhibit	O
a	O
growth	O
and	O
survival	O
advantage	O
over	O
their	O
parental	B-Cell
cells	I-Cell
of	O
origin	O
.	O

Membrane	O
-	O
anchored	O
Cbl	B-Gene_or_gene_product
suppresses	O
Hck	B-Gene_or_gene_product
protein	O
-	O
tyrosine	B-Amino_acid
kinase	O
mediated	O
cellular	B-Cell
transformation	O
.	O

The	O
mammalian	B-Organism
proto	O
-	O
oncogene	O
Cbl	B-Gene_or_gene_product
and	O
its	O
cellular	B-Cell
homologues	O
in	O
Caenorhabditis	B-Organism
elegans	I-Organism
(	O
Sli	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
and	O
Drosophila	B-Organism
(	O
D	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Cbl	I-Gene_or_gene_product
)	O
are	O
negative	O
regulators	O
of	O
some	O
growth	B-Gene_or_gene_product
factor	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
signaling	O
pathways	O
.	O

Herein	O
we	O
show	O
that	O
Cbl	B-Gene_or_gene_product
can	O
negatively	O
regulate	O
another	O
signaling	O
molecule	O
,	O
namely	O
theSrc	B-Gene_or_gene_product
-	O
family	O
kinase	O
Hck	B-Gene_or_gene_product
by	O
targeting	O
it	O
for	O
degradation	O
.	O

Hck	B-Gene_or_gene_product
-	O
mediated	O
cellular	B-Cell
transformation	O
of	O
murine	B-Organism
fibroblasts	B-Cell
is	O
reverted	O
by	O
ectopic	O
expression	O
of	O
a	O
membrane	B-Cellular_component
-	O
anchored	O
allele	O
of	O
Cbl	B-Gene_or_gene_product
as	O
assessed	O
by	O
the	O
cellular	B-Cell
morphology	O
,	O
suppression	O
of	O
anchorage	O
independent	O
growth	O
,	O
and	O
an	O
overall	O
reduction	O
in	O
the	O
total	O
tyrosine	B-Amino_acid
phosphorylation	O
levels	O
within	O
the	O
cells	B-Cell
.	O

The	O
expression	O
of	O
Cbl	B-Gene_or_gene_product
at	O
the	O
plasma	B-Cellular_component
membrane	I-Cellular_component
targets	O
both	O
Hck	B-Gene_or_gene_product
and	O
itself	O
for	O
ubiquitination	O
and	O
degradation	O
,	O
requiring	O
an	O
intact	O
RING	O
finger	O
.	O

Pharmacological	O
inhibition	O
of	O
the	O
proteasome	O
prevents	O
the	O
degradation	O
of	O
Hck	B-Gene_or_gene_product
correlating	O
with	O
an	O
increase	O
in	O
the	O
phosphotyrosine	B-Amino_acid
levels	O
within	O
the	O
cells	B-Cell
.	O

Activated	O
Hck	B-Gene_or_gene_product
and	O
membrane	B-Cellular_component
-	O
anchored	O
Cbl	B-Gene_or_gene_product
are	O
present	O
in	O
similar	O
subcellular	B-Cellular_component
localizations	O
and	O
co	O
-	O
immunoprecipitate	O
,	O
suggesting	O
that	O
their	O
interaction	O
is	O
required	O
for	O
subsequent	O
ubiquitination	O
and	O
degradation	O
.	O

Interestingly	O
,	O
both	O
constitutively	O
active	O
and	O
kinase	O
-	O
inactive	O
Hck	B-Gene_or_gene_product
interact	O
with	O
and	O
are	O
targeted	O
for	O
degradation	O
by	O
Cbl	B-Gene_or_gene_product
.	O

This	O
work	O
illustrates	O
alternate	O
means	O
to	O
regulate	O
Src	B-Gene_or_gene_product
-	O
family	O
kinases	O
,	O
and	O
suggests	O
that	O
Cbl	B-Gene_or_gene_product
may	O
be	O
able	O
to	O
suppress	O
many	O
signaling	O
pathways	O
that	O
are	O
activated	O
in	O
various	O
proliferative	O
syndromes	O
including	O
cancer	B-Cancer
.	O

Multiple	O
stages	O
of	O
malignant	O
transformation	O
of	O
human	B-Organism
endothelial	B-Cell
cells	I-Cell
modelled	O
by	O
co	O
-	O
expression	O
of	O
telomerase	B-Gene_or_gene_product
reverse	I-Gene_or_gene_product
transcriptase	I-Gene_or_gene_product
,	O
SV40	B-Gene_or_gene_product
T	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
and	O
oncogenic	O
N	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
.	O

We	O
have	O
modelled	O
multiple	O
stages	O
of	O
malignant	O
transformation	O
of	O
human	B-Organism
endothelial	B-Cell
cells	I-Cell
(	O
ECs	B-Cell
)	O
by	O
overexpressing	O
the	O
catalytic	O
subunit	O
of	O
human	B-Organism
telomerase	B-Gene_or_gene_product
(	O
hTERT	B-Gene_or_gene_product
)	O
,	O
together	O
with	O
SV40	B-Organism
T	B-Gene_or_gene_product
antigen	I-Gene_or_gene_product
(	O
SV40T	B-Gene_or_gene_product
)	O
and	O
oncogenic	O
N	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
.	O

Transfection	O
with	O
hTERT	B-Gene_or_gene_product
alone	O
,	O
led	O
to	O
the	O
immortalization	O
of	O
two	O
out	O
of	O
three	O
cultures	O
of	O
bone	B-Cell
marrow	I-Cell
-	I-Cell
derived	I-Cell
ECs	I-Cell
(	O
BMECs	B-Cell
)	O
.	O

One	O
hTERT	B-Gene_or_gene_product
transduced	O
BMEC	B-Cell
culture	O
underwent	O
a	O
long	O
proliferative	O
lag	O
before	O
resuming	O
proliferation	O
.	O

BMECs	B-Cell
transfected	O
with	O
hTERT	B-Gene_or_gene_product
alone	O
were	O
functionally	O
and	O
phenotypically	O
normal	O
.	O

BMECs	B-Cell
transfected	O
with	O
SV40T	B-Gene_or_gene_product
(	O
BMSVTs	B-Cell
)	O
had	O
an	O
extended	O
lifespan	O
,	O
but	O
eventually	O
succumbed	O
to	O
crisis	O
.	O

BMSVTs	B-Cell
exhibited	O
a	O
partially	O
transformed	O
phenotype	O
,	O
demonstrating	O
growth	O
factor	O
independence	O
,	O
altered	O
antigen	O
expression	O
and	O
forming	O
tiny	O
,	O
infrequent	O
colonies	O
in	O
vitro	O
.	O

Transduction	O
of	O
BMSVTs	B-Cell
with	O
hTERT	B-Gene_or_gene_product
resulted	O
in	O
immortalization	O
of	O
4	O
out	O
of	O
4	O
cultures	B-Cell
.	O

BMSVTs	B-Cell
immortalized	O
with	O
hTERT	B-Gene_or_gene_product
formed	O
large	O
colonies	O
in	O
vitro	O
and	O
small	O
transient	O
tumours	B-Cancer
in	O
vivo	O
.	O

BMECs	B-Cell
co	O
-	O
expressing	O
SV40T	B-Gene_or_gene_product
,	O
hTERT	B-Gene_or_gene_product
and	O
N	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
exhibited	O
an	O
overtly	O
transformed	O
phenotype	O
;	O
forming	O
very	O
large	O
colonies	O
with	O
an	O
altered	O
morphology	O
and	O
generating	O
rapidly	O
growing	O
tumours	B-Cancer
in	O
vivo	O
.	O

These	O
investigations	O
demonstrate	O
transformation	O
of	O
human	B-Organism
ECs	B-Cell
to	O
an	O
overtly	O
malignant	O
phenotype	O
.	O

This	O
model	O
will	O
be	O
useful	O
for	O
understanding	O
mechanisms	O
underlying	O
vascular	B-Pathological_formation
and	O
angiogenic	B-Pathological_formation
neoplasias	I-Pathological_formation
,	O
as	O
well	O
as	O
for	O
testing	O
drugs	O
designed	O
to	O
curtail	O
aberrant	O
EC	B-Cell
growth	O
.	O

Retinal	B-Multi-tissue_structure
microangiopathies	O
overlying	O
pigment	B-Tissue
epithelial	I-Tissue
detachment	O
in	O
age	O
-	O
related	O
macular	B-Tissue
degeneration	O
.	O

PURPOSE	O
:	O
To	O
evaluate	O
alterations	O
in	O
the	O
retinal	B-Multi-tissue_structure
vasculature	I-Multi-tissue_structure
overlying	O
pigment	B-Pathological_formation
epithelial	I-Pathological_formation
detachments	I-Pathological_formation
(	O
PED	B-Pathological_formation
)	O
in	O
exudative	O
age	O
-	O
related	O
macular	B-Tissue
degeneration	O
(	O
ARMD	O
)	O
using	O
indocyanine	B-Simple_chemical
green	I-Simple_chemical
and	O
fluorescein	B-Simple_chemical
angiography	O
.	O

METHODS	O
:	O
Forty	O
-	O
one	O
patients	B-Organism
(	O
41	O
eyes	B-Organ
)	O
with	O
a	O
clinical	O
diagnosis	O
of	O
exudative	O
ARMD	O
with	O
PED	B-Pathological_formation
underwent	O
simultaneous	O
fluorescein	B-Simple_chemical
and	O
indocyanine	B-Simple_chemical
green	I-Simple_chemical
angiography	O
,	O
also	O
under	O
high	O
(	O
10	O
degrees	O
)	O
magnification	O
.	O

Vascular	B-Multi-tissue_structure
abnormalities	O
in	O
the	O
retina	B-Multi-tissue_structure
were	O
compared	O
between	O
patients	B-Organism
with	O
vascularized	O
(	O
n	O
=	O
34	O
,	O
group	O
1	O
)	O
and	O
nonvascularized	O
(	O
n	O
=	O
7	O
,	O
group	O
2	O
)	O
PED	B-Pathological_formation
on	O
indocyanine	B-Simple_chemical
green	I-Simple_chemical
angiography	O
and	O
correlated	O
with	O
the	O
size	O
of	O
the	O
PED	B-Pathological_formation
and	O
the	O
presence	O
of	O
serous	B-Multi-tissue_structure
retinal	I-Multi-tissue_structure
detachment	O
.	O

RESULTS	O
:	O
In	O
all	O
,	O
67	O
vascular	B-Multi-tissue_structure
abnormalities	O
were	O
found	O
by	O
indocyanine	B-Simple_chemical
green	I-Simple_chemical
angiography	O
and	O
only	O
22	O
by	O
fluorescein	B-Simple_chemical
angiography	O
;	O
this	O
finding	O
was	O
statistically	O
significant	O
(	O
P	O
less	O
than	O
0	O
.	O
0001	O
)	O
.	O

The	O
finding	O
of	O
retinal	B-Multi-tissue_structure
vasculopathy	O
(	O
32	O
patients	B-Organism
in	O
group	O
1	O
and	O
two	O
patients	B-Organism
in	O
group	O
2	O
)	O
was	O
directly	O
correlated	O
with	O
the	O
presence	O
of	O
choroidal	B-Multi-tissue_structure
neovascularizations	O
(	O
P	O
=	O
0	O
.	O
002	O
)	O
.	O

There	O
was	O
also	O
a	O
direct	O
correlation	O
between	O
the	O
presence	O
of	O
choroidal	B-Multi-tissue_structure
neovascularization	O
and	O
size	O
of	O
the	O
PED	B-Pathological_formation
(	O
P	O
=	O
0	O
.	O
03	O
)	O
.	O

The	O
number	O
of	O
retinal	B-Multi-tissue_structure
vascular	I-Multi-tissue_structure
findings	O
was	O
not	O
significantly	O
correlated	O
with	O
serous	B-Organism_substance
elevation	O
of	O
the	O
retina	B-Multi-tissue_structure
.	O

CONCLUSIONS	O
:	O
Retinal	B-Multi-tissue_structure
vasculopathies	O
may	O
be	O
observed	O
in	O
eyes	B-Organ
with	O
PED	B-Pathological_formation
and	O
are	O
detectable	O
by	O
indocyanine	B-Simple_chemical
green	I-Simple_chemical
and	O
fluorescein	B-Simple_chemical
angiography	O
.	O

Fas	B-Gene_or_gene_product
-	O
Fas	B-Gene_or_gene_product
ligand	I-Gene_or_gene_product
signaling	O
pathway	O
mediates	O
an	O
interleukin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
12	I-Gene_or_gene_product
-	O
induced	O
rejection	O
of	O
a	O
murine	B-Organism
prostate	B-Cancer
tumor	I-Cancer
system	O
.	O

BACKGROUND	O
:	O
Recent	O
data	O
suggest	O
that	O
anti	O
-	O
tumor	B-Cancer
activities	O
of	O
interleukin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
12	I-Gene_or_gene_product
(	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
12	I-Gene_or_gene_product
)	O
involve	O
the	O
induction	O
of	O
apoptosis	O
.	O

Fas	B-Gene_or_gene_product
(	O
APO	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
/	O
CD95	B-Gene_or_gene_product
)	O
is	O
a	O
type	O
I	O
membrane	B-Cellular_component
protein	O
that	O
is	O
capable	O
of	O
initiating	O
an	O
apoptosis	O
signaling	O
pathway	O
when	O
bound	O
to	O
its	O
ligand	O
(	O
FasL	B-Gene_or_gene_product
)	O
.	O

We	O
undertook	O
this	O
study	O
to	O
test	O
the	O
hypothesis	O
that	O
Fas	B-Gene_or_gene_product
-	O
FasL	B-Gene_or_gene_product
-	O
mediated	O
apoptosis	O
plays	O
a	O
role	O
in	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
12	I-Gene_or_gene_product
-	O
induced	O
tumor	B-Cancer
regression	O
.	O

METHODS	O
:	O
An	O
mIL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
12	I-Gene_or_gene_product
expression	O
vector	O
driven	O
by	O
cytomegalovirus	B-Organism
promoter	O
was	O
used	O
to	O
express	O
murine	B-Organism
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
12	I-Gene_or_gene_product
cDNA	O
in	O
the	O
RM	B-Cell
-	I-Cell
9	I-Cell
murine	I-Cell
prostate	I-Cell
carcinoma	I-Cell
cell	I-Cell
line	I-Cell
.	O

Control	O
RM	B-Cell
-	I-Cell
9	I-Cell
cells	I-Cell
and	O
RM	B-Cell
-	I-Cell
9	I-Cell
cells	I-Cell
stably	O
transfected	O
with	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
12	I-Gene_or_gene_product
gene	O
(	O
RM	O
-	O
9	O
-	O
IL12	B-Gene_or_gene_product
)	O
were	O
inoculated	O
subcutaneously	B-Immaterial_anatomical_entity
in	O
4	O
-	O
to	O
6	O
-	O
week	O
-	O
old	O
male	O
C57BL	B-Organism
/	I-Organism
J6	I-Organism
mice	I-Organism
.	O

Tumor	B-Cancer
size	O
was	O
measured	O
every	O
3	O
days	O
.	O

Western	O
blot	O
and	O
immunohistochemical	O
assays	O
were	O
used	O
to	O
evaluate	O
Fas	B-Gene_or_gene_product
and	O
FasL	B-Gene_or_gene_product
protein	O
expression	O
.	O

In	O
situ	O
fluorescent	O
end	O
labeling	O
was	O
used	O
to	O
label	O
apoptotic	O
cells	B-Cell
.	O

RESULTS	O
:	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
12	I-Gene_or_gene_product
-	O
expressing	O
RM	B-Cell
-	I-Cell
9	I-Cell
prostate	I-Cell
carcinoma	I-Cell
cells	I-Cell
transplanted	O
into	O
C57BL	B-Organism
/	I-Organism
J6	I-Organism
mice	I-Organism
grew	O
more	O
slowly	O
than	O
control	O
RM	B-Cell
-	I-Cell
9	I-Cell
cells	I-Cell
and	O
vector	O
control	O
RM	B-Cell
-	I-Cell
9	I-Cell
-	I-Cell
Luc	I-Cell
cells	I-Cell
.	O

The	O
average	O
survival	O
time	O
of	O
the	O
RM	B-Organism
-	I-Organism
9	I-Organism
-	I-Organism
IL12	I-Organism
mice	I-Organism
was	O
longer	O
than	O
53	O
days	O
,	O
whereas	O
the	O
mean	O
survival	O
for	O
mice	B-Organism
transplanted	O
with	O
control	O
RM	B-Cell
-	I-Cell
9	I-Cell
cells	I-Cell
was	O
only	O
16	O
days	O
.	O

Apoptotic	O
cells	B-Cell
were	O
more	O
numerous	O
in	O
RM	B-Cancer
-	I-Cancer
9	I-Cancer
-	I-Cancer
IL12	I-Cancer
tumors	I-Cancer
:	O
10	O
.	O
3	O
%	O
vs	O
.	O
1	O
.	O
5	O
%	O
in	O
control	O
(	O
P	O
=	O
0	O
.	O
001	O
)	O
.	O

Fas	B-Gene_or_gene_product
and	O
FasL	B-Gene_or_gene_product
proteins	O
were	O
increased	O
approximately	O
twofold	O
in	O
the	O
RM	B-Cancer
-	I-Cancer
9	I-Cancer
-	I-Cancer
IL12	I-Cancer
tumors	I-Cancer
compared	O
with	O
the	O
RM	B-Cancer
-	I-Cancer
9	I-Cancer
control	I-Cancer
tumors	I-Cancer
as	O
determined	O
by	O
Western	O
blot	O
and	O
immunohistochemical	O
analyses	O
(	O
P	O
<	O
0	O
.	O
05	O
)	O
.	O

CONCLUSION	O
:	O
The	O
Fas	B-Gene_or_gene_product
-	O
FasL	B-Gene_or_gene_product
-	O
mediated	O
apoptosis	O
pathway	O
may	O
contribute	O
to	O
the	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
12	I-Gene_or_gene_product
-	O
induced	O
rejection	O
of	O
prostate	B-Cancer
carcinoma	I-Cancer
.	O

Adaptor	O
protein	O
Crk	B-Gene_or_gene_product
is	O
required	O
for	O
ephrin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
B1	I-Gene_or_gene_product
-	O
induced	O
membrane	B-Cellular_component
ruffling	O
and	O
focal	B-Cellular_component
complex	I-Cellular_component
assembly	O
of	O
human	B-Organism
aortic	B-Cell
endothelial	I-Cell
cells	I-Cell
.	O

Endothelial	B-Cell
cell	I-Cell
migration	O
is	O
an	O
essential	O
step	O
in	O
vasculogenesis	O
and	O
angiogenesis	O
,	O
in	O
which	O
receptor	B-Gene_or_gene_product
tyrosine	I-Gene_or_gene_product
kinases	I-Gene_or_gene_product
play	O
a	O
pivotal	O
role	O
.	O

We	O
investigated	O
the	O
mechanism	O
by	O
which	O
ephrin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
B1	I-Gene_or_gene_product
promotes	O
membrane	B-Cellular_component
ruffling	O
in	O
human	B-Organism
aortic	B-Cell
endothelial	I-Cell
cells	I-Cell
,	O
because	O
membrane	B-Cellular_component
ruffling	O
heralds	O
cell	B-Cell
body	I-Cell
migration	O
.	O

We	O
especially	O
focused	O
on	O
the	O
role	O
of	O
Crk	B-Gene_or_gene_product
adaptor	O
protein	O
in	O
EphB	B-Gene_or_gene_product
-	O
mediated	O
signaling	O
.	O

Using	O
DsRed	B-Simple_chemical
-	O
tagged	O
Crk	B-Gene_or_gene_product
and	O
a	O
fluorescent	O
time	O
-	O
lapse	O
microscope	O
,	O
we	O
showed	O
that	O
Crk	B-Gene_or_gene_product
was	O
recruited	O
to	O
the	O
nascent	B-Cellular_component
focal	I-Cellular_component
complex	I-Cellular_component
after	O
ephrin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
B1	I-Gene_or_gene_product
stimulation	O
.	O

Furthermore	O
,	O
we	O
found	O
that	O
p130	B-Gene_or_gene_product
(	I-Gene_or_gene_product
Cas	I-Gene_or_gene_product
)	I-Gene_or_gene_product
,	O
but	O
not	O
paxillin	B-Gene_or_gene_product
,	O
recruited	O
Crk	B-Gene_or_gene_product
to	O
the	O
nascent	B-Cellular_component
focal	I-Cellular_component
complex	I-Cellular_component
.	O

The	O
necessity	O
of	O
Crk	B-Gene_or_gene_product
in	O
ephrin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
B1	I-Gene_or_gene_product
-	O
induced	O
membrane	B-Cellular_component
ruffling	O
was	O
shown	O
both	O
by	O
the	O
overexpression	O
of	O
dominant	O
negative	O
Crk	B-Gene_or_gene_product
mutants	O
and	O
by	O
the	O
depletion	O
of	O
Crk	B-Gene_or_gene_product
by	O
using	O
RNA	O
interference	O
.	O

Then	O
,	O
we	O
examined	O
the	O
role	O
of	O
two	O
major	O
downstream	O
molecules	O
of	O
Crk	B-Gene_or_gene_product
,	O
Rac1	B-Gene_or_gene_product
and	O
Rap1	B-Gene_or_gene_product
.	O

The	O
dominant	O
negative	O
mutant	O
of	O
Rac1	B-Gene_or_gene_product
completely	O
inhibited	O
ephrin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
B1	I-Gene_or_gene_product
-	O
induced	O
membrane	B-Cellular_component
ruffling	O
and	O
focal	B-Cellular_component
complex	I-Cellular_component
assembly	O
.	O

In	O
contrast	O
,	O
rap1GAPII	B-Gene_or_gene_product
,	O
a	O
negative	O
regulator	O
of	O
Rap1	B-Gene_or_gene_product
,	O
did	O
not	O
inhibit	O
ephrin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
B1	I-Gene_or_gene_product
-	O
induced	O
membrane	B-Cellular_component
ruffling	O
.	O

However	O
,	O
in	O
rap1GAPII	B-Gene_or_gene_product
-	O
expressing	O
cells	O
,	O
ephrin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
B1	I-Gene_or_gene_product
did	O
not	O
induce	O
membrane	B-Cellular_component
spreading	O
,	O
probably	O
due	O
to	O
instability	O
of	O
the	O
focal	B-Cellular_component
complex	I-Cellular_component
.	O

These	O
results	O
indicated	O
that	O
Crk	B-Gene_or_gene_product
plays	O
a	O
critical	O
role	O
in	O
Rac1	B-Gene_or_gene_product
-	O
induced	O
membrane	B-Cellular_component
ruffling	O
and	O
Rap1	B-Gene_or_gene_product
-	O
mediated	O
nascent	B-Cellular_component
focal	I-Cellular_component
complex	I-Cellular_component
stabilization	O
contributing	O
to	O
ephrin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
B1	I-Gene_or_gene_product
-	O
induced	O
human	B-Organism
aortic	B-Cell
endothelial	I-Cell
cells	I-Cell
migration	O
.	O

Postoperative	O
progression	O
of	O
pulmonary	B-Organ
metastasis	O
in	O
osteosarcoma	B-Cancer
.	O

Early	O
relapse	O
with	O
distant	O
metastasis	O
often	O
is	O
observed	O
in	O
patients	B-Organism
with	O
cancer	B-Cancer
after	O
resection	O
of	O
the	O
primary	O
tumor	B-Cancer
.	O

It	O
is	O
considered	O
that	O
resection	O
of	O
the	O
primary	O
tumor	B-Cancer
induces	O
activation	O
of	O
systemic	O
angiogenesis	O
and	O
enhances	O
progression	O
of	O
remote	O
metastasis	O
.	O

The	O
authors	O
show	O
that	O
resection	O
of	O
the	O
primary	O
osteosarcoma	B-Cancer
tumor	I-Cancer
enhances	O
progression	O
of	O
pulmonary	B-Organ
metastasis	O
in	O
animal	O
osteosarcoma	B-Cancer
models	O
.	O

Matrigel	O
plug	O
neovascularization	O
assay	O
revealed	O
that	O
systemic	O
angiogenic	O
activity	O
was	O
elevated	O
after	O
primary	O
tumor	B-Cancer
removal	O
(	O
tumor	B-Cancer
intact	O
group	O
,	O
1	O
.	O
61	O
+	O
/	O
-	O
0	O
.	O
21	O
g	O
/	O
dL	O
;	O
tumor	B-Cancer
removed	O
group	O
,	O
4	O
.	O
92	O
+	O
/	O
-	O
0	O
.	O
35	O
g	O
/	O
dL	O
)	O
.	O

In	O
addition	O
,	O
serum	B-Organism_substance
concentration	O
of	O
the	O
angiogenesis	O
inhibitor	O
,	O
endostatin	B-Gene_or_gene_product
,	O
decreased	O
significantly	O
after	O
primary	O
tumor	B-Cancer
removal	O
.	O

Treatment	O
with	O
the	O
antiangiogenic	O
reagent	O
TNP	B-Simple_chemical
-	I-Simple_chemical
470	I-Simple_chemical
suppressed	O
postoperative	O
progression	O
of	O
pulmonary	B-Organ
metastasis	O
.	O

These	O
results	O
indicate	O
the	O
possibility	O
that	O
activation	O
of	O
angiogenic	O
activity	O
after	O
resection	O
of	O
osteosarcoma	B-Cancer
tumors	I-Cancer
enhances	O
progression	O
of	O
pulmonary	B-Organ
metastasis	O
.	O

The	O
current	O
data	O
also	O
suggest	O
that	O
administration	O
of	O
antiangiogenic	O
reagents	O
can	O
prevent	O
progression	O
of	O
pulmonary	B-Organ
metastasis	O
in	O
osteosarcoma	B-Cancer
postoperatively	O
.	O

Antiapoptotic	O
effect	O
of	O
coagulation	B-Gene_or_gene_product
factor	I-Gene_or_gene_product
VIIa	I-Gene_or_gene_product
.	O

Binding	O
of	O
factor	B-Gene_or_gene_product
VIIa	I-Gene_or_gene_product
(	O
FVIIa	B-Gene_or_gene_product
)	O
to	O
its	O
cellular	B-Cell
receptor	O
tissue	B-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
TF	B-Gene_or_gene_product
)	O
was	O
previously	O
shown	O
to	O
induce	O
various	O
intracellular	B-Immaterial_anatomical_entity
signaling	O
events	O
,	O
which	O
were	O
thought	O
to	O
be	O
responsible	O
for	O
TF	B-Gene_or_gene_product
-	O
mediated	O
biologic	O
effects	O
,	O
including	O
angiogenesis	O
,	O
tumor	B-Cancer
metastasis	O
,	O
and	O
restenosis	O
.	O

To	O
understand	O
the	O
mechanisms	O
behind	O
these	O
processes	O
,	O
we	O
have	O
examined	O
the	O
effect	O
of	O
FVIIa	B-Gene_or_gene_product
on	O
apoptosis	O
.	O

Serum	B-Organism_substance
deprivation	O
-	O
induced	O
apoptosis	O
of	O
BHK	B-Cell
(	I-Cell
+	I-Cell
TF	I-Cell
)	I-Cell
cells	I-Cell
was	O
characterized	O
by	O
apoptotic	O
blebs	B-Cellular_component
,	O
nuclei	B-Cellular_component
with	O
chromatin	B-Cellular_component
-	O
condensed	O
bodies	O
,	O
DNA	B-Cellular_component
degradation	O
,	O
and	O
activation	O
of	O
caspase	B-Gene_or_gene_product
3	I-Gene_or_gene_product
.	O

FVIIa	B-Gene_or_gene_product
markedly	O
decreased	O
the	O
number	O
of	O
cells	B-Cell
with	O
apoptotic	O
morphology	O
and	O
prevented	O
the	O
DNA	B-Cellular_component
degradation	O
as	O
measured	O
by	O
means	O
of	O
TdT	B-Gene_or_gene_product
-	O
mediated	O
dUTP	B-Simple_chemical
nick	O
end	O
labeling	O
(	O
TUNEL	O
)	O
.	O

The	O
antiapoptotic	O
effect	O
of	O
FVIIa	B-Gene_or_gene_product
was	O
confirmed	O
by	O
the	O
observation	O
that	O
FVIIa	B-Gene_or_gene_product
attenuated	O
caspase	B-Gene_or_gene_product
3	I-Gene_or_gene_product
activation	O
.	O

FVIIa	B-Gene_or_gene_product
-	O
induced	O
antiapoptotic	O
effect	O
was	O
dependent	O
on	O
its	O
proteolytic	O
activity	O
and	O
TF	B-Gene_or_gene_product
but	O
independent	O
of	O
factor	B-Gene_or_gene_product
Xa	I-Gene_or_gene_product
and	O
thrombin	B-Gene_or_gene_product
.	O

FVIIa	B-Gene_or_gene_product
-	O
induced	O
cell	B-Cell
survival	O
correlated	O
with	O
the	O
activation	O
of	O
Akt	B-Gene_or_gene_product
and	O
was	O
inhibited	O
markedly	O
by	O
the	O
specific	O
PI3	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
inhibitor	O
,	O
LY294002	B-Simple_chemical
.	O

Blocking	O
the	O
activation	O
of	O
p44	B-Gene_or_gene_product
/	I-Gene_or_gene_product
42	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
by	O
the	O
specific	O
mitogen	B-Gene_or_gene_product
-	I-Gene_or_gene_product
induced	I-Gene_or_gene_product
extracellular	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
(	O
MEK	B-Gene_or_gene_product
)	O
inhibitor	O
,	O
U0126	B-Simple_chemical
,	O
impaired	O
modestly	O
the	O
ability	O
of	O
FVIIa	B-Gene_or_gene_product
to	O
promote	O
cell	B-Cell
survival	O
.	O

In	O
conclusion	O
,	O
FVIIa	B-Gene_or_gene_product
binding	O
to	O
TF	B-Gene_or_gene_product
provided	O
protection	O
against	O
apoptosis	O
induced	O
by	O
growth	O
factor	O
deprivation	O
,	O
primarily	O
through	O
activation	O
of	O
PI3	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
/	O
Akt	B-Gene_or_gene_product
pathway	O
,	O
and	O
to	O
a	O
lesser	O
extent	O
,	O
p44	B-Gene_or_gene_product
/	I-Gene_or_gene_product
42	I-Gene_or_gene_product
MAPK	I-Gene_or_gene_product
pathway	O
.	O

Cloning	O
and	O
characterization	O
of	O
the	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
of	O
a	O
specific	O
Epstein	B-Organism
-	I-Organism
Barr	I-Organism
virus	I-Organism
variant	O
derived	O
from	O
the	O
nasopharyngeal	B-Cancer
carcinoma	I-Cancer
in	O
the	O
Taiwanese	O
population	O
.	O

A	O
DNA	O
fragment	O
containing	O
Epstein	B-Organism
-	I-Organism
Barr	I-Organism
virus	I-Organism
(	O
EBV	B-Organism
)	O
terminal	O
fragment	O
sequence	O
was	O
obtained	O
from	O
a	O
genomic	O
library	O
of	O
nasopharyngeal	B-Cancer
carcinoma	I-Cancer
(	O
NPC	B-Cancer
)	O
.	O

One	O
of	O
the	O
clones	O
(	O
clone	O
1510	O
)	O
contained	O
the	O
gene	O
encoding	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

Sequence	O
analysis	O
revealed	O
that	O
this	O
gene	O
had	O
95	O
%	O
homology	O
with	O
the	O
LMP	B-Gene_or_gene_product
sequence	O
of	O
the	O
B95	B-Organism
-	I-Organism
8	I-Organism
strain	O
.	O

Among	O
the	O
sequence	O
variations	O
,	O
there	O
was	O
a	O
change	O
from	O
G	O
to	O
T	O
at	O
nucleotide	O
position	O
169	O
,	O
426	O
,	O
resulting	O
in	O
the	O
loss	O
of	O
an	O
XhoI	B-Gene_or_gene_product
site	O
in	O
exon	O
1	O
of	O
the	O
LMP	B-Gene_or_gene_product
gene	O
.	O

A	O
pair	O
of	O
primers	O
bracketing	O
the	O
XhoI	B-Gene_or_gene_product
site	O
were	O
designed	O
to	O
synthesize	O
the	O
EBV	B-Organism
DNA	B-Cellular_component
fragment	O
from	O
nucleotides	O
169	O
,	O
081	O
-	O
169	O
,	O
577	O
by	O
using	O
the	O
polymerase	O
chain	O
reaction	O
(	O
PCR	O
)	O
method	O
.	O

The	O
PCR	O
products	O
were	O
then	O
subject	O
to	O
XhoI	B-Gene_or_gene_product
digestion	O
and	O
to	O
DNA	B-Cellular_component
sequencing	O
analysis	O
.	O

This	O
restriction	O
enzyme	O
site	O
polymorphism	O
along	O
with	O
the	O
sequence	O
variations	O
were	O
also	O
observed	O
in	O
50	O
biopsy	B-Tissue
tissues	I-Tissue
as	O
well	O
as	O
in	O
the	O
throat	B-Organism_subdivision
washings	O
of	O
6	O
out	O
of	O
20	O
healthy	O
individuals	B-Organism
that	O
we	O
examined	O
,	O
indicating	O
that	O
the	O
EBV	B-Organism
strain	O
predominantly	O
existing	O
in	O
these	O
biopsy	B-Tissue
tissues	I-Tissue
was	O
different	O
from	O
strains	O
of	O
B95	B-Organism
-	I-Organism
8	I-Organism
,	O
Jijoye	B-Organism
or	O
nude	B-Organism
mouse	I-Organism
passaged	O
cells	B-Cell
(	O
C15	B-Cell
)	O
with	O
an	O
African	O
origin	O
,	O
but	O
closely	O
resembled	O
other	O
nude	B-Organism
mouse	I-Organism
passaged	O
CAO	B-Cell
cells	I-Cell
which	O
were	O
originally	O
derived	O
from	O
China	O
.	O

Balb	B-Cell
/	I-Cell
c	I-Cell
3T3	I-Cell
cells	I-Cell
carrying	O
this	O
NPC	B-Cancer
-	O
LMP	B-Gene_or_gene_product
gene	O
showed	O
a	O
transformed	O
cell	B-Cell
morphology	O
and	O
were	O
tumorigenic	O
in	O
nude	B-Organism
mice	I-Organism
.	O

The	O
relationship	O
between	O
this	O
unique	O
type	O
of	O
EBV	B-Organism
and	O
NPC	B-Cancer
has	O
yet	O
to	O
be	O
established	O
.	O

Angiopoietin	B-Gene_or_gene_product
/	O
tie	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
as	O
mediators	O
of	O
angiogenesis	O
:	O
a	O
role	O
in	O
congestive	O
heart	B-Organ
failure	O
?	O

Angiogenic	O
factors	O
,	O
in	O
particular	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
the	O
angiopoietins	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
,	O
have	O
recently	O
generated	O
significant	O
interest	O
,	O
especially	O
in	O
oncology	O
.	O

The	O
process	O
of	O
angiogenesis	O
is	O
also	O
thought	O
to	O
occur	O
in	O
response	O
to	O
ischaemic	O
conditions	O
,	O
which	O
lie	O
at	O
the	O
core	O
of	O
cardiovascular	B-Anatomical_system
disease	O
states	O
such	O
as	O
coronary	B-Multi-tissue_structure
artery	I-Multi-tissue_structure
disease	O
and	O
congestive	O
heart	B-Organ
failure	O
.	O

However	O
,	O
current	O
data	O
do	O
not	O
conclusively	O
show	O
evidence	O
of	O
angiogenesis	O
per	O
se	O
in	O
these	O
conditions	O
,	O
despite	O
(	O
for	O
example	O
)	O
the	O
presence	O
of	O
high	O
levels	O
of	O
VEGF	B-Gene_or_gene_product
and	O
Ang	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
.	O

High	O
levels	O
of	O
these	O
angiogenic	O
factors	O
in	O
heart	B-Organ
disease	O
also	O
have	O
not	O
translated	O
into	O
clinically	O
significant	O
new	O
vessel	B-Multi-tissue_structure
formation	O
,	O
as	O
in	O
accelerated	O
cancer	B-Cancer
growth	O
or	O
proliferative	O
retinopathy	O
.	O

Indeed	O
,	O
we	O
would	O
hypothesize	O
that	O
these	O
angiogenic	O
markers	O
-	O
-	O
especially	O
the	O
angiopoietins	B-Gene_or_gene_product
-	O
-	O
do	O
not	O
necessarily	O
translate	O
into	O
new	O
vessel	B-Multi-tissue_structure
formation	O
in	O
congestive	O
heart	B-Organ
failure	O
(	O
CHF	O
)	O
,	O
but	O
may	O
well	O
reflect	O
disturbances	O
of	O
endothelial	B-Tissue
integrity	O
in	O
CHF	O
.	O

[	O
Correlation	O
between	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
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
in	O
tumor	B-Cell
-	I-Cell
associated	I-Cell
macrophages	I-Cell
and	O
lymphatic	B-Organ
metastasis	O
in	O
oral	B-Cancer
cancer	I-Cancer
]	O
.	O

BACKGROUND	O
&	O
#	O
38	O
;	O
OBJECTIVE	O
:	O
Previous	O
study	O
shows	O
that	O
both	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	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
and	O
tumor	B-Cell
-	I-Cell
associated	I-Cell
macrophages	I-Cell
(	O
TAMs	B-Cell
)	O
are	O
related	O
to	O
lymphatic	B-Organ
metastasis	O
.	O

This	O
study	O
aimed	O
to	O
explore	O
the	O
correlation	O
between	O
the	O
expression	O
of	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
in	O
TAMs	B-Cell
and	O
lymphatic	B-Organ
metastasis	O
in	O
human	B-Organism
oral	B-Cancer
squamous	I-Cancer
-	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
(	O
OSCC	B-Cancer
)	O
.	O

METHODS	O
:	O
After	O
immunohistochemical	O
staining	O
,	O
light	O
microscope	O
was	O
used	O
for	O
counting	O
macrophages	B-Cell
and	O
automated	O
image	O
analysis	O
quantification	O
was	O
used	O
to	O
determine	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
expression	O
,	O
which	O
was	O
reflected	O
by	O
positive	O
index	O
(	O
PI	O
)	O
.	O

In	O
addition	O
,	O
the	O
double	O
staining	O
was	O
also	O
used	O
to	O
determine	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
expression	O
in	O
TAMs	B-Cell
.	O

RESULTS	O
:	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
expression	O
was	O
higher	O
in	O
lymphatic	B-Organ
metastasis	O
group	O
(	O
PI	O
=	O
12	O
.	O
169	O
+	O
/	O
-	O
2	O
.	O
778	O
)	O
than	O
in	O
no	O
-	O
metastasis	O
group	O
(	O
PI	O
=	O
8	O
.	O
498	O
+	O
/	O
-	O
2	O
.	O
674	O
,	O
P	O
<	O
0	O
.	O
05	O
)	O
.	O

TAMs	B-Cell
counts	O
was	O
related	O
to	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
expression	O
in	O
OSCC	B-Cancer
(	O
r	O
=	O
0	O
.	O
370	O
,	O
P	O
<	O
0	O
.	O
05	O
)	O
.	O

The	O
result	O
of	O
double	O
staining	O
indicated	O
that	O
macrophage	B-Cell
with	O
positive	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
expression	O
accounted	O
for	O
about	O
22	O
.	O
8	O
%	O
of	O
the	O
total	O
.	O

CONCLUSION	O
:	O
Not	O
only	O
tumor	B-Cell
cells	I-Cell
but	O
also	O
TAMs	B-Cell
secrete	O
the	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
in	O
OSCC	B-Cancer
,	O
and	O
TAMs	B-Cell
may	O
play	O
a	O
major	O
role	O
in	O
peritumoral	B-Multi-tissue_structure
lymphatic	I-Multi-tissue_structure
neoangiogenesis	O
and	O
lymphatic	B-Organ
metastasis	O
.	O

Hypothesis	O
:	O
Induced	O
angiogenesis	O
after	O
surgery	O
in	O
premenopausal	O
node	O
-	O
positive	O
breast	B-Cancer
cancer	I-Cancer
patients	B-Organism
is	O
a	O
major	O
underlying	O
reason	O
why	O
adjuvant	O
chemotherapy	O
works	O
particularly	O
well	O
for	O
those	O
patients	B-Organism
.	O

BACKGROUND	O
:	O
We	O
suggest	O
that	O
surgical	O
extirpation	O
of	O
primary	O
breast	B-Cancer
cancer	I-Cancer
among	O
other	O
effects	O
accelerates	O
relapse	O
for	O
some	O
premenopausal	O
node	O
-	O
positive	O
patients	B-Organism
.	O

These	O
accelerated	O
relapses	O
occur	O
within	O
10	O
months	O
of	O
surgery	O
for	O
untreated	O
patients	B-Organism
.	O

The	O
mechanism	O
proposed	O
is	O
a	O
stimulation	O
of	O
angiogenesis	O
for	O
distant	B-Cancer
dormant	I-Cancer
micrometastases	I-Cancer
.	O

This	O
has	O
been	O
suggested	O
as	O
one	O
of	O
the	O
mechanisms	O
to	O
explain	O
the	O
mammography	O
paradox	O
for	O
women	B-Organism
aged	O
40	O
-	O
49	O
years	O
.	O

We	O
could	O
imagine	O
that	O
it	O
also	O
plays	O
a	O
role	O
in	O
adjuvant	O
chemotherapy	O
effectiveness	O
since	O
,	O
perhaps	O
not	O
coincidentally	O
,	O
this	O
is	O
most	O
beneficial	O
for	O
premenopausal	O
node	O
-	O
positive	O
patients	B-Organism
.	O

HYPOTHESIS	O
:	O
We	O
speculate	O
that	O
there	O
is	O
a	O
burst	O
of	O
angiogenesis	O
of	O
distant	B-Cancer
dormant	I-Cancer
micrometastases	I-Cancer
after	O
surgery	O
in	O
approximately	O
20	O
%	O
of	O
premenopausal	O
node	O
-	O
positive	O
patients	B-Organism
.	O

We	O
also	O
speculate	O
that	O
this	O
synchronizes	O
them	O
into	O
a	O
temporal	O
highly	O
chemosensitive	O
state	O
and	O
is	O
the	O
underlying	O
reason	O
why	O
adjuvant	O
chemotherapy	O
works	O
particularly	O
well	O
for	O
that	O
patient	B-Organism
category	O
.	O

Furthermore	O
,	O
this	O
may	O
explain	O
why	O
cancer	B-Cancer
in	O
younger	O
patients	B-Organism
is	O
more	O
often	O
'	O
aggressive	O
'	O
.	O

TESTING	O
THE	O
HYPOTHESIS	O
:	O
Stimulation	O
of	O
dormant	B-Cancer
micrometastases	I-Cancer
by	O
primary	O
tumor	B-Cancer
removal	O
is	O
known	O
to	O
occur	O
in	O
animal	O
models	O
.	O

However	O
,	O
we	O
need	O
to	O
determine	O
whether	O
it	O
happens	O
in	O
breast	B-Cancer
cancer	I-Cancer
.	O

Transient	O
circulating	O
levels	O
of	O
angioactive	O
molecules	O
and	O
serial	O
high	O
-	O
resolution	O
imaging	O
studies	O
of	O
focal	O
angiogenesis	O
might	O
help	O
.	O

IMPLICATIONS	O
:	O
Short	O
-	O
course	O
cytotoxic	O
chemotherapy	O
after	O
surgery	O
has	O
probably	O
reached	O
its	O
zenith	O
,	O
and	O
other	O
strategies	O
,	O
perhaps	O
antiangiogenic	O
methods	O
,	O
are	O
needed	O
to	O
successfully	O
treat	O
more	O
patients	B-Organism
.	O

In	O
addition	O
,	O
the	O
hypothesis	O
predicts	O
that	O
early	O
detection	O
,	O
which	O
is	O
designed	O
to	O
find	O
more	O
patients	B-Organism
without	O
involved	O
lymph	B-Multi-tissue_structure
nodes	I-Multi-tissue_structure
,	O
may	O
not	O
be	O
a	O
synergistic	O
strategy	O
with	O
adjuvant	O
chemotherapy	O
,	O
which	O
works	O
best	O
with	O
positive	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
patients	B-Organism
.	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
in	O
seizures	O
:	O
a	O
double	O
-	O
edged	O
sword	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
vascular	O
growth	O
factor	O
which	O
induces	O
angiogenesis	O
(	O
the	O
development	O
of	O
new	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
)	O
,	O
vascular	B-Multi-tissue_structure
permeability	O
,	O
and	O
inflammation	O
.	O

In	O
brain	B-Organ
,	O
receptors	O
for	O
VEGF	B-Gene_or_gene_product
have	O
been	O
localized	O
to	O
vascular	B-Tissue
endothelium	I-Tissue
,	O
neurons	B-Cell
,	O
and	O
glia	B-Cell
.	O

VEGF	B-Gene_or_gene_product
is	O
upregulated	O
after	O
hypoxic	O
injury	O
to	O
the	O
brain	B-Organ
,	O
which	O
can	O
occur	O
during	O
cerebral	B-Organ
ischemia	O
or	O
high	O
-	O
altitude	O
edema	B-Pathological_formation
,	O
and	O
has	O
been	O
implicated	O
in	O
the	O
blood	B-Multi-tissue_structure
-	I-Multi-tissue_structure
brain	I-Multi-tissue_structure
barrier	I-Multi-tissue_structure
breakdown	O
associated	O
with	O
these	O
conditions	O
.	O

Given	O
its	O
recently	O
-	O
described	O
role	O
as	O
an	O
inflammatory	O
mediator	O
,	O
VEGF	B-Gene_or_gene_product
could	O
also	O
contribute	O
to	O
the	O
inflammatory	O
responses	O
observed	O
in	O
cerebral	B-Organ
ischemia	O
.	O

After	O
seizures	O
,	O
blood	B-Multi-tissue_structure
-	I-Multi-tissue_structure
brain	I-Multi-tissue_structure
barrier	I-Multi-tissue_structure
breakdown	O
and	O
inflammation	O
is	O
also	O
observed	O
in	O
brain	B-Organ
,	O
albeit	O
on	O
a	O
lower	O
scale	O
than	O
that	O
observed	O
after	O
stroke	O
.	O

Recent	O
evidence	O
has	O
suggested	O
a	O
role	O
for	O
inflammation	O
in	O
seizure	O
disorders	O
.	O

We	O
have	O
described	O
striking	O
increases	O
in	O
VEGF	B-Gene_or_gene_product
protein	O
in	O
both	O
neurons	B-Cell
and	O
glia	B-Cell
after	O
pilocarpine	B-Simple_chemical
-	O
induced	O
status	O
epilepticus	O
in	O
the	O
brain	B-Organ
.	O

Increases	O
in	O
VEGF	B-Gene_or_gene_product
could	O
contribute	O
to	O
the	O
blood	B-Multi-tissue_structure
-	I-Multi-tissue_structure
brain	I-Multi-tissue_structure
barrier	I-Multi-tissue_structure
breakdown	O
and	O
inflammation	O
observed	O
after	O
seizures	O
.	O

However	O
,	O
VEGF	B-Gene_or_gene_product
has	O
also	O
been	O
shown	O
to	O
be	O
neuroprotective	O
across	O
several	O
experimental	O
paradigms	O
,	O
and	O
hence	O
could	O
potentially	O
protect	O
vulnerable	O
cells	B-Cell
from	O
damage	O
associated	O
with	O
seizures	O
.	O

Therefore	O
,	O
the	O
role	O
of	O
VEGF	B-Gene_or_gene_product
after	O
seizures	O
could	O
be	O
either	O
protective	O
or	O
destructive	O
.	O

Although	O
only	O
further	O
research	O
will	O
determine	O
the	O
exact	O
nature	O
of	O
VEGF	B-Gene_or_gene_product
'	O
s	O
role	O
after	O
seizures	O
,	O
preliminary	O
data	O
indicate	O
that	O
VEGF	B-Gene_or_gene_product
plays	O
a	O
protective	O
role	O
after	O
seizures	O
.	O

Safety	O
of	O
verteporfin	B-Simple_chemical
for	O
treatment	O
of	O
subfoveal	B-Multi-tissue_structure
choroidal	I-Multi-tissue_structure
neovascular	I-Multi-tissue_structure
membranes	I-Multi-tissue_structure
associated	O
with	O
age	O
-	O
related	O
macular	B-Tissue
degeneration	O
.	O

Photodynamic	O
therapy	O
(	O
PDT	O
)	O
is	O
a	O
novel	O
treatment	O
entity	O
that	O
exploits	O
the	O
photophysical	O
properties	O
of	O
various	O
photosensitive	O
chemical	O
entities	O
which	O
,	O
upon	O
light	O
activation	O
,	O
results	O
in	O
targeted	O
photooxidation	O
and	O
subsequent	O
tissue	B-Tissue
destruction	O
.	O

The	O
antiangiogenic	O
properties	O
of	O
PDT	O
have	O
been	O
adapted	O
for	O
treatment	O
of	O
subfoveal	B-Multi-tissue_structure
choroidal	I-Multi-tissue_structure
neovascular	I-Multi-tissue_structure
membranes	I-Multi-tissue_structure
due	O
to	O
disease	O
states	O
such	O
as	O
age	O
-	O
related	O
macular	B-Tissue
degeneration	O
(	O
AMD	O
)	O
.	O

Historically	O
,	O
PDT	O
has	O
been	O
limited	O
by	O
a	O
lack	O
of	O
suitable	O
photosensitive	O
dyes	O
.	O

However	O
,	O
agents	O
such	O
as	O
verteporfin	B-Simple_chemical
,	O
a	O
second	O
-	O
generation	O
benzoporphyrin	B-Simple_chemical
derivative	O
,	O
appear	O
to	O
be	O
free	O
from	O
the	O
extensive	O
phototoxicity	O
that	O
limited	O
the	O
success	O
of	O
previous	O
agents	O
.	O

Verteporfin	B-Simple_chemical
has	O
a	O
high	O
affinity	O
for	O
choroidal	B-Multi-tissue_structure
neovascular	I-Multi-tissue_structure
membranes	I-Multi-tissue_structure
,	O
typically	O
found	O
with	O
exudative	O
AMD	O
,	O
and	O
upon	O
photoactivation	O
results	O
in	O
targeted	O
microvascular	B-Tissue
damage	O
and	O
thrombus	B-Pathological_formation
formation	O
with	O
resultant	O
vessel	B-Multi-tissue_structure
occlusion	O
.	O

Scrutiny	O
of	O
diagnostic	O
indicators	O
for	O
verteporfin	B-Simple_chemical
administration	O
,	O
including	O
critical	O
angiographic	O
evaluation	O
of	O
lesion	B-Pathological_formation
size	O
and	O
visual	O
acuity	O
,	O
is	O
essential	O
to	O
treatment	O
success	O
.	O

Large	O
lesions	B-Pathological_formation
with	O
relatively	O
good	O
visual	O
acuity	O
(	O
20	O
/	O
50	O
or	O
better	O
)	O
may	O
be	O
at	O
particular	O
risk	O
for	O
marked	O
vision	O
loss	O
following	O
verteporfin	B-Simple_chemical
administration	O
.	O

Lesion	B-Pathological_formation
composition	O
also	O
appears	O
to	O
influence	O
visual	O
outcome	O
with	O
verteporfin	B-Simple_chemical
use	O
.	O

The	O
safety	O
of	O
verteporfin	B-Simple_chemical
is	O
directly	O
dependent	O
upon	O
the	O
appropriate	O
integration	O
of	O
dosage	O
,	O
infusion	O
and	O
light	O
activation	O
required	O
for	O
a	O
suitable	O
pharmacotherapeutic	O
outcome	O
.	O

When	O
used	O
appropriately	O
,	O
and	O
with	O
adequate	O
patient	B-Organism
education	O
regarding	O
photosensitivity	O
,	O
the	O
risk	O
-	O
benefit	O
of	O
verteporfin	B-Simple_chemical
for	O
the	O
medical	O
treatment	O
of	O
neovascular	O
AMD	O
is	O
favourable	O
.	O

Retrospective	O
case	O
series	O
of	O
juxtafoveal	B-Multi-tissue_structure
choroidal	I-Multi-tissue_structure
neovascularization	O
treated	O
with	O
photodynamic	O
therapy	O
with	O
verteporfin	B-Simple_chemical
.	O

PURPOSE	O
:	O
To	O
describe	O
visual	O
acuity	O
and	O
angiographic	O
outcomes	O
of	O
juxtafoveal	B-Multi-tissue_structure
choroidal	I-Multi-tissue_structure
neovascularization	O
(	O
CNV	O
)	O
treated	O
with	O
photodynamic	O
therapy	O
and	O
verteporfin	B-Simple_chemical
(	O
PDT	O
)	O
.	O

METHODS	O
:	O
Four	O
hundred	O
eighty	O
-	O
four	O
consecutive	O
eyes	B-Organ
of	O
446	O
patients	B-Organism
treated	O
with	O
PDT	O
from	O
January	O
1	O
,	O
2001	O
,	O
to	O
June	O
30	O
,	O
2002	O
,	O
were	O
identified	O
from	O
billing	O
records	O
.	O

Fluorescein	B-Simple_chemical
angiograms	O
were	O
reviewed	O
retrospectively	O
to	O
identify	O
juxtafoveal	B-Immaterial_anatomical_entity
CNV	O
.	O

Eligible	O
patients	B-Organism
had	O
CNV	O
in	O
which	O
the	O
central	O
boundary	O
of	O
the	O
lesion	B-Pathological_formation
was	O
between	O
1	O
and	O
199	O
microm	O
from	O
the	O
geometric	O
center	O
of	O
the	O
foveal	B-Tissue
avascular	I-Tissue
zone	I-Tissue
(	O
FAZ	B-Tissue
)	O
.	O

Patient	B-Organism
charts	O
were	O
reviewed	O
for	O
visual	O
acuity	O
of	O
the	O
treated	O
eye	B-Organ
before	O
PDT	O
and	O
at	O
6	O
-	O
and	O
12	O
-	O
month	O
follow	O
-	O
up	O
examinations	O
.	O

Presence	O
of	O
subfoveal	O
CNV	O
at	O
6	O
and	O
12	O
months	O
of	O
follow	O
-	O
up	O
was	O
determined	O
by	O
review	O
of	O
fluorescein	B-Simple_chemical
angiograms	O
.	O

A	O
lesion	B-Pathological_formation
was	O
considered	O
subfoveal	O
if	O
it	O
extended	O
underneath	O
the	O
geometric	O
center	O
of	O
the	O
FAZ	B-Tissue
.	O

RESULTS	O
:	O
Twenty	O
-	O
one	O
eyes	B-Organ
had	O
juxtafoveal	B-Immaterial_anatomical_entity
CNV	O
.	O

Median	O
change	O
in	O
visual	O
acuity	O
both	O
6	O
and	O
12	O
months	O
after	O
the	O
initial	O
PDT	O
was	O
0	O
lines	O
(	O
n	O
=	O
18	O
at	O
6	O
months	O
,	O
range	O
-	O
14	O
to	O
+	O
8	O
lines	O
;	O
n	O
=	O
17	O
at	O
12	O
months	O
,	O
range	O
-	O
18	O
to	O
+	O
7	O
lines	O
)	O
.	O

Eleven	O
lesions	B-Pathological_formation
progressed	O
to	O
a	O
subfoveal	O
location	O
by	O
12	O
months	O
.	O

Visual	O
acuity	O
in	O
eyes	B-Organ
with	O
progressive	O
lesions	B-Pathological_formation
decreased	O
a	O
median	O
of	O
4	O
lines	O
of	O
vision	O
.	O

CONCLUSIONS	O
:	O
Despite	O
a	O
small	O
sample	O
size	O
and	O
limited	O
length	O
of	O
follow	O
-	O
up	O
,	O
this	O
study	O
shows	O
that	O
visual	O
acuity	O
on	O
average	O
can	O
remain	O
stable	O
for	O
at	O
least	O
12	O
months	O
after	O
PDT	O
of	O
juxtafoveal	B-Pathological_formation
lesions	I-Pathological_formation
.	O

Growth	O
through	O
the	O
foveal	B-Tissue
center	I-Tissue
still	O
can	O
occur	O
,	O
however	O
,	O
and	O
this	O
can	O
be	O
associated	O
with	O
substantial	O
visual	O
loss	O
.	O

Expression	O
of	O
DCC	B-Gene_or_gene_product
and	O
netrin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
in	O
normal	O
human	B-Organism
endometrium	B-Multi-tissue_structure
and	O
its	O
implication	O
in	O
endometrial	B-Multi-tissue_structure
carcinogenesis	O
.	O

OBJECTIVE	O
:	O
Although	O
DCC	B-Gene_or_gene_product
has	O
been	O
considered	O
as	O
a	O
candidate	O
tumor	B-Cancer
suppressor	O
,	O
the	O
roles	O
it	O
plays	O
in	O
the	O
uterine	B-Multi-tissue_structure
endometrium	I-Multi-tissue_structure
and	O
in	O
the	O
carcinogenic	O
process	O
remains	O
unclear	O
.	O

To	O
define	O
these	O
roles	O
more	O
clearly	O
,	O
we	O
examined	O
the	O
expression	O
of	O
DCC	B-Gene_or_gene_product
and	O
its	O
ligand	O
,	O
netrin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
in	O
the	O
normal	O
endometrium	B-Multi-tissue_structure
and	O
in	O
endometrial	B-Cancer
cancer	I-Cancer
.	O

METHODS	O
:	O
The	O
expression	O
of	O
DCC	B-Gene_or_gene_product
and	O
netrin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
in	O
normal	O
endometrial	B-Multi-tissue_structure
glands	I-Multi-tissue_structure
and	O
in	O
cancer	B-Cell
cell	I-Cell
lines	I-Cell
was	O
examined	O
by	O
RT	O
-	O
PCR	O
and	O
immunohistochemistry	O
.	O

The	O
effects	O
of	O
exogenous	O
DCC	B-Gene_or_gene_product
and	O
netrin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
expression	O
were	O
observed	O
together	O
with	O
the	O
respective	O
expression	O
vector	O
transfection	O
.	O

RESULTS	O
:	O
Endometrial	B-Multi-tissue_structure
glands	I-Multi-tissue_structure
in	O
the	O
proliferative	O
and	O
early	O
secretory	O
phase	O
expressed	O
both	O
DCC	B-Gene_or_gene_product
and	O
netrin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
but	O
glands	B-Multi-tissue_structure
in	O
the	O
late	O
-	O
secretory	O
phase	O
tended	O
to	O
silence	O
DCC	B-Gene_or_gene_product
expression	O
.	O

In	O
addition	O
,	O
all	O
of	O
the	O
endometrial	B-Cell
cancer	I-Cell
cell	I-Cell
lines	I-Cell
lost	O
normal	O
DCC	B-Gene_or_gene_product
expression	O
.	O

Restored	O
DCC	B-Gene_or_gene_product
expression	O
in	O
the	O
cancer	B-Cell
cell	I-Cell
lines	I-Cell
in	O
the	O
absence	O
of	O
netrin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
induced	O
apoptosis	O
.	O

However	O
,	O
no	O
changes	O
were	O
observed	O
in	O
the	O
presence	O
of	O
netrin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

CONCLUSION	O
:	O
Our	O
observations	O
suggest	O
that	O
DCC	B-Gene_or_gene_product
/	O
netrin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
signaling	O
may	O
commit	O
cells	B-Cell
to	O
the	O
transition	O
of	O
endometrial	B-Multi-tissue_structure
gland	I-Multi-tissue_structure
architecture	O
or	O
function	O
from	O
a	O
proliferating	O
to	O
a	O
secretory	O
phase	O
.	O

In	O
addition	O
,	O
the	O
silencing	O
of	O
DCC	B-Gene_or_gene_product
expression	O
may	O
contribute	O
to	O
the	O
escape	O
of	O
endometrial	B-Cell
cancer	I-Cell
cells	I-Cell
from	O
a	O
DCC	B-Gene_or_gene_product
-	O
regulated	O
apoptotic	O
program	O
,	O
thereby	O
promoting	O
malignant	O
phenotypes	O
.	O

An	O
evaluation	O
of	O
low	B-Simple_chemical
molecular	I-Simple_chemical
weight	I-Simple_chemical
heparin	I-Simple_chemical
and	O
hyperbaric	B-Simple_chemical
oxygen	I-Simple_chemical
treatment	O
in	O
the	O
prevention	O
of	O
intra	O
-	O
abdominal	O
adhesions	O
and	O
wound	B-Pathological_formation
healing	O
.	O

BACKGROUND	O
:	O
Abdominal	O
surgery	O
can	O
lead	O
to	O
intra	O
-	O
abdominal	O
adhesions	O
with	O
significant	O
morbidity	O
and	O
mortality	O
.	O

To	O
prevent	O
adhesions	O
,	O
an	O
experimental	O
study	O
was	O
planned	O
to	O
designate	O
the	O
effects	O
of	O
low	B-Simple_chemical
molecular	I-Simple_chemical
weight	I-Simple_chemical
(	I-Simple_chemical
LMW	I-Simple_chemical
)	I-Simple_chemical
heparins	I-Simple_chemical
and	O
hyperbaric	B-Simple_chemical
oxygen	I-Simple_chemical
(	O
HBO	B-Simple_chemical
)	O
therapy	O
both	O
on	O
the	O
formation	O
of	O
adhesions	O
and	O
wound	B-Pathological_formation
healing	O
.	O

METHODS	O
:	O
Thirty	O
-	O
eight	O
Wistar	B-Organism
albino	I-Organism
rats	I-Organism
underwent	O
laparotomy	O
to	O
cause	O
intra	O
-	O
abdominal	O
adhesions	O
by	O
mechanical	O
abrasion	O
of	O
the	O
cecum	B-Multi-tissue_structure
and	O
ethanol	B-Simple_chemical
application	O
.	O

The	O
rats	B-Organism
were	O
divided	O
into	O
4	O
groups	O
.	O

In	O
the	O
control	O
group	O
(	O
group	O
1	O
)	O
no	O
further	O
management	O
was	O
undertaken	O
.	O

Group	O
2	O
was	O
treated	O
by	O
Enoxaparine	B-Simple_chemical
Na	I-Simple_chemical
,	O
group	O
3	O
received	O
HBO	B-Simple_chemical
therapy	O
,	O
and	O
group	O
4	O
was	O
given	O
both	O
enoxaparine	B-Simple_chemical
Na	I-Simple_chemical
and	O
HBO	B-Simple_chemical
treatment	O
.	O

RESULTS	O
:	O
There	O
was	O
a	O
statistically	O
significant	O
difference	O
between	O
the	O
control	O
and	O
enoxaparine	B-Simple_chemical
Na	I-Simple_chemical
groups	O
regarding	O
adhesions	O
.	O

Statistically	O
significant	O
differences	O
were	O
observed	O
between	O
groups	O
1	O
and	O
4	O
and	O
between	O
groups	O
1	O
and	O
3	O
regarding	O
the	O
hydroxyproline	O
content	O
of	O
the	O
abdominal	B-Pathological_formation
wounds	I-Pathological_formation
.	O

In	O
the	O
pathologic	O
analysis	O
of	O
the	O
abdominal	B-Pathological_formation
wounds	I-Pathological_formation
,	O
there	O
was	O
no	O
statistically	O
significant	O
difference	O
between	O
any	O
of	O
the	O
groups	O
,	O
including	O
the	O
control	O
group	O
,	O
regarding	O
inflammation	O
.	O

Statistically	O
significant	O
differences	O
were	O
observed	O
regarding	O
angiogenesis	O
between	O
the	O
control	O
group	O
and	O
groups	O
3	O
and	O
4	O
.	O

There	O
was	O
also	O
a	O
statistically	O
significant	O
difference	O
regarding	O
fibrosis	O
between	O
groups	O
1	O
and	O
4	O
.	O

CONCLUSIONS	O
:	O
Enoxaparine	B-Simple_chemical
Na	I-Simple_chemical
decreased	O
intra	O
-	O
abdominal	O
adhesions	O
,	O
and	O
HBO	B-Simple_chemical
therapy	O
had	O
no	O
beneficial	O
effect	O
on	O
adhesions	O
.	O

Enoxaparine	B-Simple_chemical
Na	I-Simple_chemical
had	O
no	O
harmful	O
effects	O
on	O
wound	B-Pathological_formation
healing	O
,	O
and	O
HBO	B-Simple_chemical
therapy	O
increased	O
the	O
process	O
of	O
wound	B-Pathological_formation
healing	O
.	O

VEGF	B-Gene_or_gene_product
-	O
targeted	O
therapy	O
:	O
therapeutic	O
potential	O
and	O
recent	O
advances	O
.	O

After	O
over	O
30	O
years	O
of	O
theorizing	O
,	O
the	O
use	O
of	O
angiogenesis	O
inhibitors	O
as	O
anticancer	B-Cancer
therapy	O
has	O
finally	O
moved	O
from	O
the	O
realm	O
of	O
research	O
to	O
reality	O
.	O

Normal	O
adult	O
vasculature	B-Anatomical_system
is	O
generally	O
quiescent	O
in	O
nature	O
,	O
with	O
endothelial	B-Cell
cells	I-Cell
dividing	O
approximately	O
every	O
10	O
years	O
.	O

In	O
contrast	O
,	O
the	O
growth	O
of	O
tumors	B-Cancer
requires	O
constant	O
vascular	B-Multi-tissue_structure
growth	O
and	O
remodeling	O
in	O
order	O
for	O
solid	B-Cancer
tumors	I-Cancer
to	O
grow	O
beyond	O
1	O
-	O
2	O
mm	O
(	O
3	O
)	O
in	O
size	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
and	O
its	O
receptors	O
are	O
key	O
regulators	O
of	O
the	O
process	O
of	O
angiogenesis	O
,	O
which	O
makes	O
them	O
attractive	O
therapeutic	O
targets	O
.	O

A	O
multitude	O
of	O
VEGF	B-Gene_or_gene_product
-	O
targeted	O
inhibitory	O
agents	O
are	O
currently	O
being	O
investigated	O
for	O
the	O
treatment	O
of	O
cancer	B-Cancer
.	O

This	O
review	O
article	O
focuses	O
on	O
recent	O
developments	O
in	O
the	O
use	O
of	O
angiogenesis	O
inhibitors	O
for	O
the	O
treatment	O
of	O
breast	B-Cancer
,	O
lung	B-Cancer
,	O
and	O
colorectal	B-Cancer
cancers	I-Cancer
.	O

MDM2	B-Gene_or_gene_product
as	O
a	O
predictor	O
of	O
prostate	B-Cancer
carcinoma	I-Cancer
outcome	O
:	O
an	O
analysis	O
of	O
Radiation	O
Therapy	O
Oncology	O
Group	O
Protocol	O
8610	O
.	O

BACKGROUND	O
:	O
The	O
MDM2	B-Gene_or_gene_product
oncoprotein	O
promotes	O
p53	B-Gene_or_gene_product
degradation	O
via	O
ubiquitin	B-Gene_or_gene_product
,	O
establishing	O
negative	O
feedback	O
control	O
of	O
p53	B-Gene_or_gene_product
and	O
consequently	O
affecting	O
cell	B-Cell
cycle	O
arrest	O
and	O
apoptosis	O
.	O

The	O
authors	O
evaluated	O
the	O
association	O
between	O
MDM2	B-Gene_or_gene_product
expression	O
and	O
local	O
failure	O
,	O
distant	O
metastasis	O
(	O
DM	O
)	O
,	O
cause	O
-	O
specific	O
mortality	O
,	O
and	O
overall	O
mortality	O
in	O
men	B-Organism
treated	O
in	O
Radiation	O
Therapy	O
Oncology	O
Group	O
8610	O
with	O
radiotherapy	O
,	O
with	O
or	O
without	O
androgen	B-Simple_chemical
deprivation	O
.	O

METHODS	O
:	O
Of	O
the	O
456	O
eligible	O
and	O
analyzable	O
patients	B-Organism
(	O
parent	O
cohort	O
)	O
,	O
adequate	O
archival	O
diagnostic	O
tissue	B-Tissue
specimens	I-Tissue
from	O
108	O
patients	B-Organism
were	O
available	O
for	O
MDM2	B-Gene_or_gene_product
analysis	O
(	O
MDM2	B-Gene_or_gene_product
cohort	O
)	O
.	O

Cox	O
proportional	O
hazards	O
multivariate	O
analysis	O
(	O
MVA	O
)	O
was	O
used	O
to	O
determine	O
the	O
relation	O
of	O
MDM2	B-Gene_or_gene_product
to	O
the	O
endpoints	O
.	O

MDM2	B-Gene_or_gene_product
overexpression	O
was	O
manually	O
classified	O
as	O
>	O
5	O
%	O
nuclear	B-Cellular_component
staining	O
.	O

An	O
image	O
analysis	O
system	O
was	O
also	O
used	O
to	O
quantify	O
the	O
proportion	O
of	O
tumor	B-Cancer
nuclei	B-Cellular_component
with	O
MDM2	B-Gene_or_gene_product
staining	O
(	O
ACIS	O
index	O
)	O
and	O
staining	O
intensity	O
.	O

RESULTS	O
:	O
Overexpression	O
of	O
MDM2	B-Gene_or_gene_product
by	O
manual	O
counts	O
was	O
seen	O
in	O
44	O
%	O
(	O
n	O
=	O
47	O
)	O
of	O
the	O
patients	B-Organism
.	O

In	O
the	O
manual	O
count	O
analysis	O
,	O
there	O
was	O
no	O
significant	O
relation	O
between	O
MDM2	B-Gene_or_gene_product
overexpression	O
and	O
outcome	O
.	O

The	O
ACIS	O
index	O
,	O
using	O
a	O
cutoff	O
point	O
defined	O
by	O
the	O
median	O
value	O
,	O
<	O
or	O
=	O
3	O
%	O
versus	O
>	O
3	O
%	O
,	O
was	O
related	O
to	O
5	O
-	O
year	O
DM	O
rates	O
in	O
univariate	O
analyses	O
(	O
32	O
.	O
6	O
%	O
vs	O
.	O
45	O
.	O
8	O
%	O
;	O
P	O
=	O
0	O
.	O
057	O
)	O
and	O
MVA	O
(	O
P	O
=	O
0	O
.	O
06	O
)	O
.	O

The	O
intensity	O
of	O
MDM2	B-Gene_or_gene_product
staining	O
was	O
not	O
significant	O
.	O

CONCLUSIONS	O
:	O
MDM2	B-Gene_or_gene_product
expression	O
quantified	O
by	O
image	O
analysis	O
was	O
weakly	O
associated	O
with	O
DM	O
.	O

The	O
cohort	O
examined	O
was	O
relatively	O
small	O
and	O
with	O
larger	O
patient	B-Organism
numbers	O
,	O
MDM2	B-Gene_or_gene_product
overexpression	O
may	O
emerge	O
as	O
a	O
more	O
significant	O
covariate	O
.	O

Thrombospondins	B-Gene_or_gene_product
,	O
metallo	B-Gene_or_gene_product
proteases	I-Gene_or_gene_product
and	O
thrombospondin	B-Gene_or_gene_product
receptors	I-Gene_or_gene_product
messenger	O
RNA	O
and	O
protein	O
expression	O
in	O
different	O
tumour	B-Cancer
sublines	I-Cancer
of	O
the	O
Dunning	B-Cancer
prostate	I-Cancer
cancer	I-Cancer
model	O
.	O

Thrombospondin	B-Gene_or_gene_product
is	O
a	O
potent	O
inhibitor	O
of	O
angiogenesis	O
and	O
might	O
therefore	O
be	O
important	O
in	O
controlling	O
tumour	B-Cancer
growth	O
.	O

TSP	B-Gene_or_gene_product
interacts	O
with	O
a	O
number	O
of	O
proteases	O
and	O
receptors	O
and	O
in	O
this	O
way	O
inhibits	O
stimulation	O
of	O
angiogenesis	O
.	O

An	O
earlier	O
study	O
showed	O
that	O
thrombospondin	B-Gene_or_gene_product
is	O
expressed	O
in	O
benign	B-Pathological_formation
prostatic	I-Pathological_formation
hyperplasia	I-Pathological_formation
(	O
BPH	B-Pathological_formation
)	O
and	O
high	O
-	O
grade	O
prostatic	B-Pathological_formation
intraepithelial	I-Pathological_formation
neoplasia	I-Pathological_formation
(	O
PIN	B-Pathological_formation
)	O
but	O
is	O
absent	O
in	O
prostate	B-Cancer
cancer	I-Cancer
.	O

The	O
present	O
study	O
was	O
therefore	O
designed	O
to	O
evaluate	O
the	O
expression	O
of	O
thrombospondin	B-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
2	B-Gene_or_gene_product
(	O
TSP	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
2	I-Gene_or_gene_product
)	O
,	O
TSP	B-Gene_or_gene_product
receptors	O
CD36	B-Gene_or_gene_product
and	O
CD47	B-Gene_or_gene_product
,	O
and	O
matrix	B-Gene_or_gene_product
-	I-Gene_or_gene_product
metalloproteases	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
9	B-Gene_or_gene_product
(	O
MMP	B-Gene_or_gene_product
-	O
,	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
)	O
in	O
a	O
rat	B-Organism
prostate	B-Cancer
cancer	I-Cancer
model	O
.	O

By	O
using	O
immunohistochemistry	O
,	O
Western	O
blot	O
,	O
and	O
real	O
-	O
time	O
PCR	O
the	O
expression	O
patterns	O
of	O
TSP	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
2	I-Gene_or_gene_product
,	O
CD36	B-Gene_or_gene_product
,	O
CD47	B-Gene_or_gene_product
,	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
and	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
were	O
investigated	O
in	O
normal	O
rat	B-Organism
prostate	B-Tissue
tissue	I-Tissue
and	O
five	O
malignant	B-Tissue
Dunning	I-Tissue
sublines	I-Tissue
tissue	I-Tissue
.	O

TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
mRNA	O
levels	O
were	O
decreased	O
in	O
all	O
tumours	B-Cancer
compared	O
with	O
normal	O
prostate	B-Organ
.	O

However	O
,	O
there	O
was	O
no	O
difference	O
in	O
expression	O
of	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
CD36	B-Gene_or_gene_product
mRNA	O
in	O
these	O
samples	O
.	O

MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
was	O
increased	O
with	O
malignancy	O
,	O
but	O
no	O
expression	O
of	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
was	O
seen	O
.	O

The	O
CD47	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
did	O
slightly	O
increase	O
with	O
malignancy	O
except	O
for	O
H3327	B-Cancer
.	O

The	O
results	O
showed	O
that	O
thrombospondin	B-Gene_or_gene_product
is	O
expressed	O
in	O
normal	O
prostate	B-Organ
but	O
not	O
in	O
prostate	B-Cancer
tumours	I-Cancer
in	O
a	O
rat	B-Organism
model	O
.	O

Simultaneously	O
,	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
expression	O
increases	O
with	O
malignancy	O
.	O

Bovine	B-Organism
papillomavirus	I-Organism
E7	B-Gene_or_gene_product
transformation	O
function	O
correlates	O
with	O
cellular	B-Cell
p600	B-Gene_or_gene_product
protein	O
binding	O
.	O

The	O
E7	B-Gene_or_gene_product
oncoprotein	O
of	O
bovine	B-Organism
papillomavirus	I-Organism
type	I-Organism
1	I-Organism
(	O
BPV	B-Organism
-	I-Organism
1	I-Organism
)	O
is	O
required	O
for	O
the	O
full	O
transformation	O
activity	O
of	O
the	O
virus	O
.	O

However	O
,	O
the	O
mechanism	O
by	O
which	O
E7	B-Gene_or_gene_product
contributes	O
to	O
cellular	B-Cell
transformation	O
is	O
unknown	O
.	O

To	O
address	O
this	O
question	O
,	O
we	O
used	O
the	O
proteomic	O
approach	O
of	O
tandem	O
affinity	O
purification	O
to	O
identify	O
cellular	B-Cell
proteins	O
that	O
are	O
in	O
complex	O
with	O
E7	B-Gene_or_gene_product
,	O
and	O
identified	O
the	O
600	O
-	O
kDa	O
protein	O
,	O
p600	B-Gene_or_gene_product
,	O
as	O
a	O
binding	O
partner	O
of	O
E7	B-Gene_or_gene_product
.	O

The	O
ability	O
of	O
E7	B-Gene_or_gene_product
to	O
complex	O
with	O
p600	B-Gene_or_gene_product
correlated	O
with	O
its	O
ability	O
to	O
enhance	O
anchorage	O
independence	O
of	O
BPV	B-Organism
-	O
1	O
E6	B-Gene_or_gene_product
-	O
expressing	O
cells	B-Cell
.	O

Furthermore	O
,	O
E7	B-Gene_or_gene_product
mutant	O
proteins	O
impaired	O
in	O
their	O
ability	O
to	O
bind	O
p600	B-Gene_or_gene_product
were	O
transformation	O
defective	O
.	O

Additionally	O
,	O
knockdown	O
of	O
p600	B-Gene_or_gene_product
reduced	O
transformation	O
of	O
cells	B-Cell
expressing	O
both	O
BPV	B-Organism
-	I-Organism
1	I-Organism
E6	B-Gene_or_gene_product
and	O
E7	B-Gene_or_gene_product
,	O
as	O
well	O
as	O
E6	B-Gene_or_gene_product
alone	O
,	O
suggesting	O
that	O
the	O
ability	O
of	O
E7	B-Gene_or_gene_product
to	O
transformed	O
cells	B-Cell
is	O
mediated	O
,	O
at	O
least	O
in	O
part	O
,	O
through	O
its	O
ability	O
to	O
bind	O
p600	B-Gene_or_gene_product
.	O

These	O
data	O
complement	O
work	O
that	O
shows	O
that	O
HPV16	B-Organism
E7	B-Gene_or_gene_product
also	O
interacts	O
with	O
p600	B-Gene_or_gene_product
,	O
and	O
that	O
this	O
interaction	O
correlates	O
with	O
the	O
ability	O
of	O
HPV16	B-Organism
E7	B-Gene_or_gene_product
to	O
transform	O
cells	B-Cell
.	O

These	O
studies	O
thus	O
identify	O
p600	B-Gene_or_gene_product
as	O
a	O
shared	O
target	O
of	O
the	O
E7	B-Gene_or_gene_product
proteins	O
of	O
multiple	O
papillomaviruses	B-Organism
.	O

Molecular	O
requirements	O
for	O
epithelial	B-Cell
-	O
mesenchymal	B-Cell
transition	O
during	O
tumor	B-Cancer
progression	O
.	O

Epithelial	B-Cell
-	O
mesenchymal	B-Cell
transitions	O
(	O
EMTs	O
)	O
occur	O
as	O
key	O
steps	O
during	O
embryonic	B-Developing_anatomical_structure
morphogenesis	O
,	O
and	O
are	O
now	O
implicated	O
in	O
the	O
progression	O
of	O
primary	O
tumors	B-Cancer
towards	O
metastases	O
.	O

Recent	O
advances	O
have	O
fostered	O
a	O
more	O
detailed	O
understanding	O
of	O
molecular	O
mechanisms	O
and	O
networks	O
governing	O
EMT	O
in	O
tumor	B-Cancer
progression	O
.	O

Besides	O
TGFbeta	B-Gene_or_gene_product
and	O
RTK	B-Gene_or_gene_product
/	O
Ras	B-Gene_or_gene_product
signaling	O
,	O
autocrine	O
factors	O
and	O
Wnt	B-Gene_or_gene_product
-	O
,	O
Notch	B-Gene_or_gene_product
-	O
,	O
Hedgehog	B-Gene_or_gene_product
-	O
and	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
-	O
dependent	O
pathways	O
were	O
found	O
to	O
contribute	O
to	O
EMT	B-Gene_or_gene_product
.	O

Repression	O
of	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
by	O
transcriptional	O
regulators	O
such	O
as	O
Snail	B-Gene_or_gene_product
or	O
Twist	B-Gene_or_gene_product
emerges	O
as	O
one	O
critical	O
step	O
driving	O
EMT	O
,	O
and	O
this	O
stage	O
is	O
currently	O
being	O
molecularly	O
linked	O
with	O
many	O
of	O
the	O
new	O
players	O
.	O

Increasing	O
evidence	O
suggests	O
that	O
EMT	O
plays	O
a	O
specific	O
role	O
in	O
the	O
migration	O
of	O
cells	B-Cell
from	O
a	O
primary	O
tumor	B-Cancer
into	O
the	O
circulation	O
and	O
may	O
provide	O
a	O
rationale	O
for	O
developing	O
more	O
effective	O
cancer	B-Cancer
therapies	O
.	O

Minimal	O
contribution	O
of	O
marrow	B-Cell
-	I-Cell
derived	I-Cell
endothelial	I-Cell
precursors	I-Cell
to	O
tumor	B-Multi-tissue_structure
vasculature	I-Multi-tissue_structure
.	O

During	O
embryogenesis	O
,	O
vascular	B-Cell
and	O
hemopoietic	B-Cell
cells	I-Cell
originate	O
from	O
a	O
common	O
precursor	O
,	O
the	O
hemangioblast	B-Cell
.	O

Recent	O
evidence	O
suggests	O
the	O
existence	O
of	O
endothelial	B-Cell
precursors	I-Cell
in	O
adult	O
bone	B-Cell
marrow	I-Cell
cells	I-Cell
,	O
but	O
it	O
is	O
unclear	O
whether	O
those	O
precursors	O
have	O
a	O
role	O
in	O
tumor	B-Cancer
neovascularization	O
.	O

In	O
this	O
report	O
,	O
we	O
demonstrate	O
that	O
murine	B-Organism
bone	B-Multi-tissue_structure
marrow	I-Multi-tissue_structure
contains	O
endothelial	B-Cell
progenitors	I-Cell
,	O
which	O
arise	O
from	O
a	O
cell	B-Cell
with	O
self	O
-	O
renewing	O
capacity	O
,	O
and	O
can	O
integrate	O
into	O
tumor	B-Tissue
microvasculature	I-Tissue
,	O
albeit	O
at	O
a	O
very	O
low	O
frequency	O
.	O

A	O
transgenic	O
double	O
-	O
reporter	O
strategy	O
allowed	O
us	O
to	O
demonstrate	O
definitively	O
that	O
tumor	B-Cell
bone	I-Cell
marrow	I-Cell
-	I-Cell
derived	I-Cell
endothelial	I-Cell
cells	I-Cell
arise	O
by	O
transdifferentiation	O
of	O
marrow	B-Cell
progenitors	I-Cell
rather	O
than	O
by	O
cell	B-Cell
fusion	O
.	O

Single	O
cell	B-Cell
transplants	O
showed	O
that	O
a	O
common	O
precursor	O
contributes	O
to	O
both	O
the	O
hemopoietic	B-Cell
and	O
endothelial	B-Cell
lineages	I-Cell
,	O
thus	O
demonstrating	O
the	O
presence	O
of	O
an	O
adult	O
hemangioblast	B-Cell
.	O

Furthermore	O
,	O
we	O
demonstrate	O
that	O
increased	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
-	I-Gene_or_gene_product
A	I-Gene_or_gene_product
secretion	O
by	O
tumor	B-Cell
cells	I-Cell
,	O
as	O
well	O
as	O
activation	O
of	O
VEGF	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
in	O
bone	B-Cell
marrow	I-Cell
cells	I-Cell
does	O
not	O
alter	O
the	O
mobilization	O
and	O
incorporation	O
of	O
marrow	B-Cell
-	I-Cell
derived	I-Cell
endothelial	I-Cell
progenitors	I-Cell
into	O
tumor	B-Tissue
vasculature	I-Tissue
.	O

Finally	O
,	O
in	O
human	B-Organism
umbilical	B-Cell
cord	I-Cell
blood	I-Cell
cells	I-Cell
,	O
we	O
show	O
that	O
endothelial	B-Cell
precursors	I-Cell
make	O
up	O
only	O
approximately	O
1	O
in	O
10	O
(	O
7	O
)	O
mononuclear	B-Cell
cells	I-Cell
but	O
are	O
highly	O
enriched	O
in	O
the	O
CD133	B-Gene_or_gene_product
+	O
cell	O
population	O
.	O

By	O
ruling	O
out	O
cell	B-Cell
fusion	O
,	O
we	O
clearly	O
demonstrate	O
the	O
existence	O
of	O
an	O
adult	O
hemangioblast	B-Cell
,	O
but	O
the	O
differentiation	O
of	O
marrow	B-Cell
stem	I-Cell
cells	I-Cell
toward	O
the	O
endothelial	B-Cell
lineage	I-Cell
is	O
an	O
extremely	O
rare	O
event	O
.	O

Furthermore	O
,	O
we	O
show	O
that	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
A	I-Gene_or_gene_product
stimulation	O
of	O
hemopoietic	B-Cell
cells	I-Cell
does	O
not	O
significantly	O
alter	O
this	O
process	O
.	O

Histopathological	O
development	O
of	O
gastric	B-Cancer
tumors	I-Cancer
induced	O
by	O
N	B-Simple_chemical
-	I-Simple_chemical
methyl	I-Simple_chemical
-	I-Simple_chemical
N	I-Simple_chemical
'	I-Simple_chemical
-	I-Simple_chemical
nitro	I-Simple_chemical
-	I-Simple_chemical
N	I-Simple_chemical
-	I-Simple_chemical
nitrosoguanidine	I-Simple_chemical
in	O
rats	B-Organism
.	O

The	O
development	O
of	O
carcinoma	B-Cancer
was	O
examined	O
in	O
male	O
Wistar	B-Organism
rats	I-Organism
(	O
n	O
=	O
120	O
)	O
exposed	O
to	O
N	B-Simple_chemical
-	I-Simple_chemical
methyl	I-Simple_chemical
-	I-Simple_chemical
N	I-Simple_chemical
'	I-Simple_chemical
-	I-Simple_chemical
nitro	I-Simple_chemical
-	I-Simple_chemical
N	I-Simple_chemical
-	I-Simple_chemical
nitrosoguanidine	I-Simple_chemical
(	O
MNNG	B-Simple_chemical
)	O
in	O
the	O
drinking	O
water	B-Simple_chemical
(	O
83	O
micrograms	O
/	O
ml	O
)	O
for	O
16	O
weeks	O
.	O

After	O
MNNG	B-Simple_chemical
administration	O
,	O
rats	B-Organism
were	O
investigated	O
by	O
endoscopic	O
observation	O
,	O
visualization	O
of	O
microvascular	B-Tissue
structure	O
,	O
and	O
estimation	O
of	O
lectin	B-Gene_or_gene_product
binding	O
sites	O
.	O

Changes	O
of	O
bile	B-Organism_substance
reflux	O
to	O
the	O
stomach	B-Organ
was	O
observed	O
endoscopically	O
at	O
24	O
weeks	O
as	O
well	O
as	O
the	O
development	O
of	O
gastric	B-Pathological_formation
mucosal	I-Pathological_formation
erosions	I-Pathological_formation
.	O

Protruding	O
and	O
expansive	O
ulcerating	B-Cancer
carcinomas	I-Cancer
developed	O
at	O
36	O
weeks	O
and	O
had	O
a	O
microvascular	B-Tissue
pattern	O
similar	O
to	O
that	O
of	O
human	B-Organism
adenocarcinoma	B-Cancer
.	O

Estimation	O
of	O
lectin	B-Gene_or_gene_product
binding	O
site	O
and	O
pattern	O
was	O
useful	O
to	O
evaluate	O
the	O
malignant	O
potential	O
of	O
cell	B-Cell
proliferation	O
.	O

We	O
postulate	O
that	O
endoscopic	O
observation	O
is	O
valuable	O
in	O
investigating	O
the	O
development	O
of	O
gastric	B-Cancer
carcinoma	I-Cancer
,	O
and	O
microvascular	B-Tissue
structure	O
and	O
lectin	B-Gene_or_gene_product
binding	O
pattern	O
may	O
be	O
useful	O
to	O
demonstrate	O
the	O
mechanism	O
of	O
growth	O
of	O
gastric	B-Cancer
carcinoma	I-Cancer
.	O

Decrease	O
in	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Myc	I-Gene_or_gene_product
activity	O
enhances	O
cancer	B-Cell
cell	I-Cell
sensitivity	O
to	O
vinblastine	B-Simple_chemical
.	O

The	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
oncogene	O
encodes	O
for	O
a	O
transcriptional	O
factor	O
involved	O
in	O
many	O
cellular	B-Cell
processes	O
such	O
as	O
proliferation	O
,	O
differentiation	O
and	O
apoptosis	O
.	O

According	O
to	O
these	O
different	O
functions	O
,	O
the	O
role	O
of	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Myc	I-Gene_or_gene_product
protein	O
in	O
cellular	B-Cell
sensitivity	O
to	O
anti	O
-	O
cancer	B-Cancer
drugs	O
is	O
controversial	O
.	O

We	O
defined	O
the	O
role	O
of	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Myc	I-Gene_or_gene_product
in	O
cancer	B-Cell
cell	I-Cell
sensitivity	O
to	O
vinblastine	B-Simple_chemical
(	O
VLB	B-Simple_chemical
)	O
using	O
human	B-Organism
colon	B-Cell
cancer	I-Cell
cells	I-Cell
:	O
LoVo	B-Cell
wild	O
-	O
type	O
or	O
transfected	O
with	O
a	O
plasmid	O
containing	O
the	O
human	B-Organism
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
gene	O
in	O
antisense	O
orientation	O
(	O
LoVo	B-Cell
-	I-Cell
mycANS	I-Cell
)	O
.	O

Analysis	O
of	O
VLB	B-Simple_chemical
cytotoxicity	O
demonstrated	O
a	O
3	O
-	O
fold	O
increase	O
in	O
VLB	B-Simple_chemical
sensitivity	O
in	O
LoVo	B-Cell
-	I-Cell
mycANS	I-Cell
cells	I-Cell
.	O

Comparison	O
between	O
cells	B-Cell
revealed	O
different	O
apoptosis	O
kinetics	O
:	O
accumulation	O
of	O
cells	B-Cell
in	O
sub	O
-	O
G1	O
phase	O
and	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
occurred	O
earlier	O
in	O
LoVo	B-Cell
-	I-Cell
mycANS	I-Cell
.	O

Then	O
,	O
we	O
demonstrated	O
a	O
mitochondrial	B-Cellular_component
membrane	I-Cellular_component
potential	O
disruption	O
followed	O
by	O
cytochrome	B-Gene_or_gene_product
c	I-Gene_or_gene_product
release	O
that	O
indicates	O
the	O
involvement	O
of	O
mitochondria	B-Cellular_component
in	O
this	O
apoptotic	O
signaling	O
pathway	O
.	O

This	O
earlier	O
apoptosis	O
was	O
accompanied	O
by	O
a	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
decrease	O
and	O
a	O
p53	B-Gene_or_gene_product
increase	O
.	O

In	O
conclusion	O
,	O
the	O
decrease	O
in	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Myc	I-Gene_or_gene_product
expression	O
enhanced	O
the	O
VLB	B-Simple_chemical
sensitivity	O
,	O
triggering	O
earlier	O
apoptosis	O
through	O
induction	O
of	O
the	O
intrinsic	O
pathway	O
.	O

Thus	O
,	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
induction	O
is	O
a	O
resistance	O
factor	O
and	O
our	O
findings	O
suggest	O
that	O
tumors	B-Cancer
carrying	O
low	O
levels	O
of	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Myc	I-Gene_or_gene_product
protein	O
could	O
be	O
more	O
responsive	O
to	O
vinca	B-Simple_chemical
alkaloids	I-Simple_chemical
treatment	O
.	O

Moreover	O
,	O
the	O
downregulation	O
of	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
oncogene	O
by	O
an	O
antisense	O
strategy	O
might	O
represent	O
a	O
useful	O
goal	O
for	O
improving	O
the	O
efficacy	O
of	O
this	O
anti	O
-	O
neoplastic	O
drug	O
family	O
.	O

Alteration	O
of	O
homeobox	B-Gene_or_gene_product
gene	O
expression	O
by	O
N	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
transformation	O
of	O
PA	B-Cell
-	I-Cell
1	I-Cell
human	I-Cell
teratocarcinoma	I-Cell
cells	I-Cell
.	O

We	O
used	O
a	O
series	O
of	O
cell	B-Cell
clones	I-Cell
from	O
a	O
human	B-Organism
teratocarcinoma	B-Cell
cell	I-Cell
line	I-Cell
,	O
PA	B-Cell
-	I-Cell
1	I-Cell
,	O
to	O
study	O
the	O
effect	O
of	O
transformation	O
by	O
an	O
activated	O
N	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
oncogene	O
on	O
the	O
expression	O
of	O
genes	O
involved	O
in	O
retinoic	B-Simple_chemical
acid	I-Simple_chemical
(	O
RA	B-Simple_chemical
)	O
-	O
induced	O
differentiation	O
and	O
growth	O
regulation	O
.	O

Recently	O
,	O
it	O
has	O
been	O
shown	O
that	O
expression	O
of	O
human	B-Organism
HOX	B-Gene_or_gene_product
2	I-Gene_or_gene_product
genes	O
is	O
sequentially	O
activated	O
by	O
RA	B-Simple_chemical
beginning	O
from	O
Hox	B-Gene_or_gene_product
2	I-Gene_or_gene_product
.	I-Gene_or_gene_product
9	I-Gene_or_gene_product
at	O
the	O
3	O
'	O
end	O
of	O
the	O
HOX	B-Gene_or_gene_product
2	I-Gene_or_gene_product
cluster	O
(	O
A	O
.	O
Simeone	O
,	O
D	O
.	O
Acampora	O
,	O
L	O
.	O
Arcioni	O
,	O
P	O
.	O
W	O
.	O
Andrews	O
,	O
E	O
.	O
Boncinelli	O
,	O
and	O
F	O
.	O
Mavilio	O
,	O
Nature	O
[	O
London	O
]	O
346	O
:	O
763	O
-	O
766	O
,	O
1990	O
)	O
.	O

We	O
now	O
report	O
that	O
six	O
different	O
genes	O
of	O
the	O
cluster	O
HOX	B-Gene_or_gene_product
1	I-Gene_or_gene_product
are	O
sequentially	O
induced	O
by	O
RA	B-Simple_chemical
in	O
a	O
similar	O
temporal	O
pattern	O
,	O
beginning	O
with	O
genes	O
at	O
the	O
3	O
'	O
end	O
of	O
the	O
cluster	O
.	O

However	O
,	O
in	O
N	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
-	O
transformed	O
cell	B-Cell
clones	I-Cell
,	O
RA	B-Simple_chemical
-	O
induced	O
expression	O
of	O
these	O
homeobox	B-Gene_or_gene_product
genes	O
is	O
delayed	O
.	O

Hox	B-Gene_or_gene_product
1	I-Gene_or_gene_product
.	I-Gene_or_gene_product
4	I-Gene_or_gene_product
and	O
Hox	B-Gene_or_gene_product
1	I-Gene_or_gene_product
.	I-Gene_or_gene_product
3	I-Gene_or_gene_product
,	O
genes	O
abundantly	O
induced	O
in	O
nontransformed	O
clones	B-Cell
after	O
3	O
days	O
of	O
RA	B-Simple_chemical
treatment	O
,	O
are	O
expressed	O
in	O
N	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
-	O
transformed	O
cells	B-Cell
only	O
after	O
10	O
days	O
of	O
RA	B-Simple_chemical
treatment	O
.	O

At	O
this	O
time	O
,	O
the	O
cells	B-Cell
'	O
growth	O
is	O
arrested	O
at	O
very	O
high	O
density	O
,	O
and	O
no	O
differentiated	O
morphologic	O
characteristics	O
are	O
observed	O
.	O

Constitutive	O
expression	O
of	O
a	O
transfected	O
Hox	B-Gene_or_gene_product
1	I-Gene_or_gene_product
.	I-Gene_or_gene_product
4	I-Gene_or_gene_product
gene	O
under	O
the	O
control	O
of	O
a	O
simian	B-Organism
virus	I-Organism
40	I-Organism
promotor	O
leads	O
to	O
differentiated	O
cell	B-Cell
morphology	O
similar	O
to	O
that	O
of	O
the	O
RA	B-Simple_chemical
-	O
induced	O
phenotype	O
and	O
restores	O
the	O
growth	O
-	O
inhibitory	O
effects	O
of	O
RA	B-Simple_chemical
in	O
N	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
-	O
transformed	O
cells	B-Cell
.	O

These	O
observations	O
provide	O
evidence	O
that	O
enhanced	O
proliferation	O
in	O
N	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
-	O
transformed	O
cells	B-Cell
compromises	O
teratocarcinoma	B-Cell
cell	I-Cell
differentiation	O
by	O
a	O
mechanism	O
that	O
transiently	O
suppresses	O
homeobox	B-Gene_or_gene_product
gene	O
induction	O
and	O
implies	O
a	O
central	O
role	O
for	O
homeobox	B-Gene_or_gene_product
genes	O
in	O
RA	B-Simple_chemical
-	O
induced	O
cell	B-Cell
differentiation	O
.	O

We	O
conclude	O
that	O
stimulation	O
of	O
a	O
putative	O
growth	O
factor	O
signal	O
pathway	O
,	O
associated	O
with	O
ras	B-Gene_or_gene_product
-	O
induced	O
proliferation	O
,	O
transiently	O
suppresses	O
the	O
induction	O
of	O
transcription	O
factors	O
functionally	O
involved	O
in	O
cell	B-Cell
growth	O
and	O
differentiation	O
.	O

High	O
-	O
Dose	O
celecoxib	B-Simple_chemical
and	O
metronomic	O
"	O
low	O
-	O
dose	O
"	O
cyclophosphamide	B-Simple_chemical
is	O
an	O
effective	O
and	O
safe	O
therapy	O
in	O
patients	B-Organism
with	O
relapsed	O
and	O
refractory	O
aggressive	O
histology	O
non	B-Cancer
-	I-Cancer
Hodgkin	I-Cancer
'	I-Cancer
s	I-Cancer
lymphoma	I-Cancer
.	O

PURPOSE	O
:	O
Angiogenesis	O
is	O
increased	O
in	O
aggressive	O
histology	O
non	B-Cancer
-	I-Cancer
Hodgkin	I-Cancer
'	I-Cancer
s	I-Cancer
lymphoma	I-Cancer
and	O
may	O
be	O
a	O
target	O
with	O
selective	O
cyclooxygenase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
inhibition	O
and	O
metronomic	O
chemotherapy	O
.	O

EXPERIMENTAL	O
DESIGN	O
:	O
We	O
assessed	O
response	O
,	O
toxicity	O
,	O
and	O
biomarkers	O
of	O
angiogenesis	O
to	O
low	O
-	O
dose	O
cyclophosphamide	B-Simple_chemical
(	O
50	O
mg	O
p	O
.	O
o	O
.	O
o	O
.	O
d	O
.	O
)	O
and	O
high	O
-	O
dose	O
celecoxib	B-Simple_chemical
(	O
400	O
mg	O
p	O
.	O
o	O
.	O
b	O
.	O
i	O
.	O
d	O
.	O
)	O
in	O
adult	O
patients	B-Organism
with	O
relapsed	O
or	O
refractory	O
aggressive	O
non	B-Cancer
-	I-Cancer
Hodgkin	I-Cancer
'	I-Cancer
s	I-Cancer
lymphoma	I-Cancer
in	O
a	O
multicenter	O
phase	O
II	O
prospective	O
study	O
.	O

RESULTS	O
:	O
Thirty	O
-	O
two	O
of	O
35	O
patients	B-Organism
(	O
median	O
age	O
,	O
62	O
years	O
)	O
are	O
evaluable	O
for	O
response	O
.	O

Patients	B-Organism
had	O
primarily	O
relapsed	O
diffuse	O
large	O
B	B-Cancer
-	I-Cancer
cell	I-Cancer
lymphoma	I-Cancer
(	O
63	O
%	O
)	O
were	O
heavily	O
pretreated	O
(	O
median	O
of	O
three	O
regimens	O
)	O
and	O
high	O
risk	O
(	O
79	O
%	O
international	O
prognostic	O
index	O
,	O
greater	O
than	O
or	O
=	O
2	O
)	O
and	O
34	O
%	O
were	O
relapsed	O
after	O
autologous	O
stem	B-Cell
cell	I-Cell
transplant	O
.	O

With	O
a	O
median	O
follow	O
-	O
up	O
of	O
8	O
.	O
4	O
months	O
,	O
the	O
overall	O
best	O
response	O
rate	O
is	O
37	O
%	O
(	O
2	O
complete	O
clinical	O
response	O
/	O
complete	O
clinical	O
response	O
unconfirmed	O
and	O
9	O
partial	O
response	O
)	O
,	O
with	O
22	O
%	O
achieving	O
stable	O
disease	O
.	O

Median	O
overall	O
and	O
progression	O
-	O
free	O
survivals	O
are	O
14	O
.	O
4	O
and	O
4	O
.	O
7	O
months	O
,	O
respectively	O
.	O

The	O
median	O
response	O
duration	O
was	O
8	O
.	O
2	O
months	O
.	O

The	O
most	O
common	O
toxicity	O
was	O
skin	B-Organ
rash	O
(	O
40	O
%	O
)	O
;	O
myelosuppression	O
and	O
gastrointestinal	B-Organism_subdivision
side	O
effects	O
were	O
uncommon	O
.	O

Three	O
patients	B-Organism
developed	O
deep	B-Pathological_formation
vein	I-Pathological_formation
thromboses	I-Pathological_formation
and	O
two	O
heavily	O
pretreated	O
patients	B-Organism
developed	O
treatment	O
-	O
related	O
acute	B-Cancer
myelogenous	I-Cancer
leukemia	I-Cancer
or	O
myelodysplasia	B-Pathological_formation
after	O
3	O
.	O
7	O
and	O
12	O
months	O
of	O
therapy	O
.	O

Circulating	O
endothelial	B-Cell
cells	I-Cell
and	O
their	O
precursors	O
declined	O
and	O
remained	O
low	O
in	O
responders	O
,	O
whereas	O
plasma	B-Organism_substance
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
trended	O
to	O
decline	O
in	O
responding	O
patients	B-Organism
but	O
increase	O
in	O
nonresponders	O
.	O

Trough	O
celecoxib	B-Simple_chemical
levels	O
achieved	O
targeted	O
"	O
antiangiogenic	O
"	O
levels	O
.	O

CONCLUSIONS	O
:	O
Low	O
-	O
dose	O
cyclophosphamide	B-Simple_chemical
and	O
high	O
-	O
dose	O
celecoxib	B-Simple_chemical
is	O
well	O
tolerated	O
and	O
active	O
in	O
pretreated	O
aggressive	O
non	B-Cancer
-	I-Cancer
Hodgkin	I-Cancer
'	I-Cancer
s	I-Cancer
lymphoma	I-Cancer
.	O

Close	O
surveillance	O
for	O
arterial	B-Multi-tissue_structure
and	O
venous	B-Multi-tissue_structure
thrombotic	O
events	O
is	O
recommended	O
.	O

The	O
decline	O
in	O
circulating	O
endothelial	B-Cell
cells	I-Cell
and	O
their	O
precursors	O
suggests	O
that	O
this	O
combination	O
may	O
be	O
working	O
by	O
inhibiting	O
angiogenesis	O
but	O
should	O
be	O
validated	O
in	O
a	O
larger	O
patient	B-Organism
sample	O
.	O

Photodynamic	O
therapy	O
with	O
verteporfin	B-Simple_chemical
for	O
subfoveal	B-Multi-tissue_structure
choroidal	I-Multi-tissue_structure
neovascularization	O
secondary	O
to	O
pathologic	O
myopia	O
:	O
long	O
-	O
term	O
study	O
.	O

PURPOSE	O
:	O
To	O
assess	O
the	O
safety	O
and	O
effectiveness	O
of	O
photodynamic	O
therapy	O
(	O
PDT	O
)	O
with	O
verteporfin	B-Simple_chemical
for	O
subfoveal	B-Multi-tissue_structure
choroidal	I-Multi-tissue_structure
neovascularization	O
(	O
CNV	O
)	O
secondary	O
to	O
pathologic	O
myopia	O
(	O
PM	O
)	O
.	O

METHODS	O
:	O
Sixty	O
-	O
two	O
patients	B-Organism
(	O
62	O
eyes	B-Organ
)	O
with	O
PM	O
underwent	O
PDT	O
according	O
to	O
the	O
guidelines	O
of	O
the	O
Verteporfin	B-Simple_chemical
in	O
Photodynamic	O
Therapy	O
Study	O
.	O

Clinical	O
evaluations	O
performed	O
at	O
all	O
study	O
visits	O
included	O
measurement	O
of	O
best	O
-	O
corrected	O
Snellen	O
visual	O
acuity	O
,	O
slit	O
-	O
lamp	O
biomicroscopy	O
,	O
and	O
fundus	B-Multi-tissue_structure
fluorescein	B-Simple_chemical
angiography	O
.	O

Patients	B-Organism
were	O
followed	O
up	O
at	O
1	O
month	O
and	O
3	O
months	O
after	O
treatment	O
and	O
thereafter	O
at	O
3	O
-	O
month	O
intervals	O
.	O

RESULTS	O
:	O
The	O
final	O
visual	O
acuity	O
of	O
the	O
study	O
patients	B-Organism
,	O
after	O
a	O
median	O
follow	O
-	O
up	O
of	O
31	O
months	O
,	O
improved	O
by	O
greater	O
than	O
or	O
=	O
1	O
Snellen	O
lines	O
in	O
8	O
patients	B-Organism
(	O
13	O
%	O
)	O
,	O
deteriorated	O
in	O
20	O
(	O
32	O
%	O
)	O
,	O
and	O
remained	O
stable	O
in	O
34	O
(	O
55	O
%	O
)	O
.	O

The	O
baseline	O
visual	O
acuity	O
was	O
similar	O
in	O
the	O
various	O
study	O
groups	O
.	O

The	O
final	O
mean	O
visual	O
acuity	O
in	O
group	O
A	O
(	O
55	O
years	O
of	O
age	O
or	O
younger	O
)	O
was	O
20	O
/	O
80	O
and	O
significantly	O
(	O
P	O
=	O
0	O
.	O
006	O
)	O
better	O
than	O
that	O
(	O
20	O
/	O
138	O
)	O
in	O
group	O
B	O
(	O
older	O
than	O
55	O
years	O
of	O
age	O
)	O
.	O

The	O
mean	O
final	O
visual	O
acuity	O
in	O
eyes	B-Organ
with	O
higher	O
refractive	O
error	O
at	O
baseline	O
(	O
greater	O
than	O
-	O
17	O
diopters	O
)	O
was	O
significantly	O
better	O
(	O
P	O
=	O
0	O
.	O
014	O
)	O
than	O
that	O
in	O
eyes	B-Organ
with	O
lower	O
refractive	O
error	O
(	O
-	O
6	O
to	O
-	O
10	O
diopters	O
)	O
.	O

CNV	O
size	O
did	O
not	O
affect	O
visual	O
outcomes	O
.	O

CONCLUSION	O
:	O
PDT	O
preserves	O
vision	O
in	O
patients	B-Organism
with	O
CNV	O
associated	O
with	O
PM	O
.	O

Younger	O
patients	B-Organism
and	O
eyes	B-Organ
with	O
higher	O
refractive	O
error	O
appear	O
more	O
likely	O
to	O
benefit	O
from	O
PDT	O
with	O
verteporfin	B-Simple_chemical
.	O

Carcinogenesis	O
and	O
transcriptional	O
regulation	O
through	O
Maf	B-Gene_or_gene_product
recognition	O
elements	O
.	O

Many	O
studies	O
on	O
carcinogenesis	O
carried	O
out	O
early	O
in	O
the	O
last	O
century	O
are	O
united	O
on	O
the	O
consensus	O
that	O
cancer	B-Cancer
is	O
a	O
genetic	O
disease	O
.	O

Cancer	B-Cell
cells	I-Cell
typically	O
display	O
gene	O
dysfunction	O
and	O
endogenous	O
or	O
exogenous	O
insults	O
resulting	O
in	O
gene	O
dysfunction	O
are	O
often	O
carcinogenic	O
.	O

Recent	O
advances	O
in	O
stem	B-Cell
cell	I-Cell
biology	O
added	O
the	O
new	O
concept	O
that	O
cancer	B-Cancer
originates	O
from	O
a	O
single	O
cancer	B-Cancer
-	O
initiating	O
cell	B-Cell
.	O

To	O
understand	O
the	O
molecular	O
basis	O
of	O
carcinogenesis	O
from	O
the	O
beginning	O
to	O
the	O
full	O
acquirement	O
of	O
malignancy	O
,	O
factors	O
concerned	O
with	O
carcinogenesis	O
were	O
categorized	O
into	O
three	O
groups	O
:	O
those	O
guarding	O
and	O
stabilizing	O
genomes	B-Cellular_component
,	O
those	O
regulating	O
cell	B-Cell
proliferation	O
,	O
and	O
those	O
conferring	O
resistance	O
to	O
various	O
micro	O
-	O
environmental	O
stresses	O
.	O

One	O
example	O
of	O
particular	O
interest	O
is	O
the	O
Keap1	B-Gene_or_gene_product
-	O
Nrf2	B-Gene_or_gene_product
system	O
since	O
,	O
according	O
to	O
recent	O
studies	O
,	O
it	O
has	O
turned	O
out	O
to	O
be	O
ambivalent	O
.	O

Nrf2	B-Gene_or_gene_product
heterodimerizes	O
with	O
small	O
Maf	B-Gene_or_gene_product
protein	O
to	O
strongly	O
activate	O
transcription	O
through	O
the	O
Maf	B-Gene_or_gene_product
recognition	O
element	O
(	O
MARE	O
)	O
and	O
Keap1	B-Gene_or_gene_product
is	O
an	O
inhibitory	O
regulator	O
of	O
Nrf2	B-Gene_or_gene_product
.	O

The	O
genes	O
regulated	O
by	O
Nrf2	B-Gene_or_gene_product
are	O
very	O
important	O
for	O
cellular	B-Cell
protection	O
of	O
the	O
genome	B-Cellular_component
from	O
xenobiotic	O
and	O
oxidative	O
stresses	O
and	O
,	O
consequently	O
,	O
for	O
preventing	O
carcinogenesis	O
.	O

This	O
implies	O
that	O
enhancing	O
Nrf2	B-Gene_or_gene_product
activity	O
is	O
a	O
promising	O
method	O
for	O
thwarting	O
cancer	B-Cancer
.	O

On	O
the	O
contrary	O
,	O
the	O
constitutive	O
activation	O
of	O
Nrf2	B-Gene_or_gene_product
due	O
to	O
mutations	O
in	O
the	O
keap1	B-Gene_or_gene_product
gene	O
is	O
characteristically	O
observed	O
in	O
lung	B-Cell
cancer	I-Cell
cells	I-Cell
,	O
suggesting	O
that	O
induced	O
expression	O
of	O
Nrf2	B-Gene_or_gene_product
target	O
genes	O
favors	O
the	O
prevalence	O
of	O
cancer	B-Cell
cells	I-Cell
.	O

Angiotensin	B-Gene_or_gene_product
II	I-Gene_or_gene_product
induces	O
soluble	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
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
release	O
via	O
calcineurin	B-Gene_or_gene_product
signaling	O
pathway	O
in	O
pregnancy	O
.	O

Maternal	O
endothelial	B-Cell
dysfunction	O
in	O
preeclampsia	O
is	O
associated	O
with	O
increased	O
soluble	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
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
sFlt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
,	O
a	O
circulating	O
antagonist	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
placental	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
.	O

Angiotensin	B-Gene_or_gene_product
II	I-Gene_or_gene_product
(	O
Ang	B-Gene_or_gene_product
II	I-Gene_or_gene_product
)	O
is	O
a	O
potent	O
vasoconstrictor	O
that	O
increases	O
concomitant	O
with	O
sFlt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
during	O
pregnancy	O
.	O

Therefore	O
,	O
we	O
speculated	O
that	O
Ang	B-Gene_or_gene_product
II	I-Gene_or_gene_product
may	O
promote	O
the	O
expression	O
of	O
sFlt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
in	O
pregnancy	O
.	O

Here	O
we	O
report	O
that	O
infusion	O
of	O
Ang	B-Gene_or_gene_product
II	I-Gene_or_gene_product
significantly	O
increases	O
circulating	O
levels	O
of	O
sFlt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
in	O
pregnant	O
mice	B-Organism
,	O
thereby	O
demonstrating	O
that	O
Ang	B-Gene_or_gene_product
II	I-Gene_or_gene_product
is	O
a	O
regulator	O
of	O
sFlt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
secretion	O
in	O
vivo	O
.	O

Furthermore	O
,	O
Ang	B-Gene_or_gene_product
II	I-Gene_or_gene_product
stimulated	O
sFlt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
production	O
in	O
a	O
dose	O
-	O
and	O
time	O
-	O
dependent	O
manner	O
from	O
human	B-Organism
villous	O
explants	O
and	O
cultured	O
trophoblasts	B-Cell
but	O
not	O
from	O
endothelial	B-Cell
cells	I-Cell
,	O
suggesting	O
that	O
trophoblasts	B-Cell
are	O
the	O
primary	O
source	O
of	O
sFlt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
during	O
pregnancy	O
.	O

As	O
expected	O
,	O
Ang	B-Gene_or_gene_product
II	I-Gene_or_gene_product
-	O
induced	O
sFlt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
secretion	O
resulted	O
in	O
the	O
inhibition	O
of	O
endothelial	B-Cell
cell	I-Cell
migration	O
and	O
in	O
vitro	O
tube	B-Tissue
formation	O
.	O

In	O
vitro	O
and	O
in	O
vivo	O
studies	O
with	O
losartan	B-Simple_chemical
,	O
small	O
interfering	O
RNA	O
specific	O
for	O
calcineurin	B-Gene_or_gene_product
and	O
FK506	B-Simple_chemical
demonstrated	O
that	O
Ang	B-Gene_or_gene_product
II	I-Gene_or_gene_product
-	O
mediated	O
sFlt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
release	O
was	O
via	O
Ang	B-Gene_or_gene_product
II	I-Gene_or_gene_product
type	I-Gene_or_gene_product
1	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
activation	O
and	O
calcineurin	O
signaling	O
,	O
respectively	O
.	O

These	O
findings	O
reveal	O
a	O
previously	O
unrecognized	O
regulatory	O
role	O
for	O
Ang	B-Gene_or_gene_product
II	I-Gene_or_gene_product
on	O
sFlt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
expression	O
in	O
murine	B-Organism
and	O
human	B-Organism
pregnancy	O
and	O
suggest	O
that	O
elevated	O
sFlt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
levels	O
in	O
preeclampsia	O
may	O
be	O
caused	O
by	O
a	O
dysregulation	O
of	O
the	O
local	O
renin	B-Gene_or_gene_product
/	O
angiotensin	B-Gene_or_gene_product
system	O
.	O

p21	B-Gene_or_gene_product
delays	O
tumor	B-Cancer
onset	O
by	O
preservation	O
of	O
chromosomal	B-Cellular_component
stability	O
.	O

The	O
p53	B-Gene_or_gene_product
protein	O
suppresses	O
tumorigenesis	O
by	O
initiating	O
cellular	B-Cell
functions	O
such	O
as	O
cell	B-Cell
cycle	O
arrest	O
and	O
apoptosis	O
in	O
response	O
to	O
DNA	B-Cellular_component
damage	O
.	O

A	O
p53	B-Gene_or_gene_product
mutant	O
,	O
p53R172P	B-Gene_or_gene_product
,	O
which	O
is	O
deficient	O
for	O
apoptosis	O
but	O
retains	O
a	O
partial	O
cell	B-Cell
cycle	O
arrest	O
function	O
,	O
delays	O
tumor	B-Cancer
onset	O
in	O
mice	B-Organism
.	O

Remarkably	O
,	O
lymphomas	B-Cancer
arising	O
in	O
Trp53	B-Gene_or_gene_product
(	O
515C	O
/	O
515C	O
)	O
mice	O
(	O
encoding	O
p53R172P	B-Gene_or_gene_product
)	O
retain	O
stable	O
genomes	O
.	O

Given	O
the	O
dominant	O
role	O
of	O
p21	B-Gene_or_gene_product
in	O
p53	B-Gene_or_gene_product
cell	B-Cell
cycle	O
control	O
,	O
we	O
crossed	O
Trp53	B-Gene_or_gene_product
(	O
515C	O
/	O
515C	O
)	O
mice	O
onto	O
a	O
p21	B-Gene_or_gene_product
-	O
null	O
background	O
to	O
determine	O
whether	O
p21	B-Gene_or_gene_product
was	O
required	O
for	O
maintaining	O
chromosomal	B-Cellular_component
stability	O
and	O
delaying	O
tumor	B-Cancer
onset	O
.	O

Loss	O
of	O
p21	B-Gene_or_gene_product
completely	O
abolished	O
the	O
cell	B-Cell
cycle	O
arrest	O
function	O
of	O
p53R172P	B-Gene_or_gene_product
and	O
accelerated	O
tumor	B-Cancer
onset	O
in	O
Trp53	B-Gene_or_gene_product
(	O
515C	O
/	O
515C	O
)	O
mice	O
.	O

Cytogenetic	O
examination	O
of	O
Trp53	B-Gene_or_gene_product
(	O
515C	O
/	O
515C	O
)	O
p21	B-Gene_or_gene_product
(	O
-	O
/	O
-	O
)	O
sarcomas	B-Cancer
and	O
lymphomas	B-Cancer
revealed	O
aneuploidy	B-Pathological_formation
and	O
chromosomal	B-Pathological_formation
aberrations	I-Pathological_formation
that	O
were	O
absent	O
in	O
Trp53	B-Gene_or_gene_product
(	O
515C	O
/	O
515C	O
)	O
malignancies	B-Cancer
.	O

Thus	O
,	O
p21	B-Gene_or_gene_product
coupled	O
p53	B-Gene_or_gene_product
-	O
dependent	O
checkpoint	O
control	O
and	O
preservation	O
of	O
chromosomal	B-Cellular_component
stability	O
,	O
and	O
cooperated	O
with	O
apoptosis	O
in	O
suppressing	O
tumor	B-Cancer
onset	O
in	O
mice	B-Organism
.	O

Mechanisms	O
for	O
the	O
magnolol	B-Simple_chemical
-	O
induced	O
cell	B-Cell
death	O
of	O
CGTH	B-Cell
W	I-Cell
-	I-Cell
2	I-Cell
thyroid	I-Cell
carcinoma	I-Cell
cells	I-Cell
.	O

Magnolol	B-Simple_chemical
,	O
a	O
substance	O
purified	O
from	O
the	O
bark	B-Tissue
of	O
Magnolia	B-Organism
officialis	I-Organism
,	O
inhibits	O
cell	B-Cell
proliferation	O
and	O
induces	O
apoptosis	O
in	O
a	O
variety	O
of	O
cancer	B-Cell
cells	I-Cell
.	O

The	O
aim	O
of	O
this	O
study	O
was	O
to	O
study	O
the	O
effects	O
of	O
magnolol	B-Simple_chemical
on	O
CGTH	B-Cell
W	I-Cell
-	I-Cell
2	I-Cell
thyroid	I-Cell
carcinoma	I-Cell
cells	I-Cell
.	O

After	O
24	O
h	O
treatment	O
with	O
80	O
microM	O
magnolol	B-Simple_chemical
in	O
serum	B-Organism_substance
-	O
containing	O
medium	O
,	O
about	O
50	O
%	O
of	O
the	O
cells	B-Cell
exhibited	O
apoptotic	O
features	O
and	O
20	O
%	O
necrotic	O
features	O
.	O

Cytochrome	B-Gene_or_gene_product
-	I-Gene_or_gene_product
c	I-Gene_or_gene_product
staining	O
was	O
diffused	O
in	O
the	O
cytoplasm	B-Organism_substance
of	O
the	O
apoptotic	O
cells	B-Cell
,	O
but	O
restricted	O
to	O
the	O
mitochondria	B-Cellular_component
in	O
control	O
cells	B-Cell
.	O

Western	O
blot	O
analyses	O
showed	O
an	O
increase	O
in	O
levels	O
of	O
activated	O
caspases	B-Gene_or_gene_product
(	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
)	O
and	O
of	O
cleaved	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
by	O
magnolol	B-Simple_chemical
.	O

Concomitantly	O
,	O
immunostaining	O
for	O
apoptosis	B-Gene_or_gene_product
inducing	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
AIF	B-Gene_or_gene_product
)	O
showed	O
a	O
time	O
-	O
dependent	O
translocation	O
from	O
the	O
mitochondria	B-Cellular_component
to	O
the	O
nucleus	B-Cellular_component
.	O

Inhibition	O
of	O
either	O
PARP	B-Gene_or_gene_product
or	O
caspase	B-Gene_or_gene_product
activity	O
blocked	O
magnolol	B-Simple_chemical
-	O
induced	O
apoptosis	O
,	O
supporting	O
the	O
involvement	O
of	O
the	O
caspases	B-Gene_or_gene_product
and	O
PARP	B-Gene_or_gene_product
.	O

In	O
addition	O
,	O
magnolol	B-Simple_chemical
activated	O
phosphatase	B-Gene_or_gene_product
and	I-Gene_or_gene_product
tensin	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
and	O
inactivated	O
Akt	B-Gene_or_gene_product
by	O
decreasing	O
levels	O
of	O
phosphorylated	O
PTEN	B-Gene_or_gene_product
and	O
phosphorylated	O
Akt	B-Gene_or_gene_product
.	O

These	O
data	O
suggest	O
that	O
magnolol	B-Simple_chemical
promoted	O
apoptosis	O
probably	O
by	O
alleviating	O
the	O
inhibitory	O
effect	O
of	O
Akt	B-Gene_or_gene_product
on	O
caspase	B-Gene_or_gene_product
9	I-Gene_or_gene_product
.	O

Furthermore	O
,	O
inhibition	O
of	O
PARP	B-Gene_or_gene_product
activity	O
,	O
but	O
not	O
of	O
caspase	B-Gene_or_gene_product
activity	O
,	O
completely	O
prevented	O
magnolol	B-Simple_chemical
-	O
induced	O
necrosis	O
,	O
suggesting	O
the	O
notion	O
that	O
it	O
might	O
be	O
caused	O
by	O
depletion	O
of	O
intracellular	B-Immaterial_anatomical_entity
ATP	B-Simple_chemical
levels	O
due	O
to	O
PARP	B-Gene_or_gene_product
activation	O
.	O

These	O
results	O
show	O
that	O
magnolol	B-Simple_chemical
initiates	O
apoptosis	O
via	O
the	O
cytochrome	B-Gene_or_gene_product
-	I-Gene_or_gene_product
c	I-Gene_or_gene_product
/	O
caspase	B-Gene_or_gene_product
3	I-Gene_or_gene_product
/	O
PARP	B-Gene_or_gene_product
/	O
AIF	B-Gene_or_gene_product
and	O
PTEN	B-Gene_or_gene_product
/	O
Akt	B-Gene_or_gene_product
/	O
caspase	B-Gene_or_gene_product
9	I-Gene_or_gene_product
/	O
PARP	B-Gene_or_gene_product
pathways	O
and	O
necrosis	O
via	O
PARP	B-Gene_or_gene_product
activation	O
.	O

Adaptive	O
landscapes	O
and	O
emergent	O
phenotypes	O
:	O
why	O
do	O
cancers	B-Cancer
have	O
high	O
glycolysis	O
?	O

Investigating	O
the	O
causes	O
of	O
increased	O
aerobic	O
glycolysis	O
in	O
tumors	B-Cancer
(	O
Warburg	O
Effect	O
)	O
has	O
gone	O
in	O
and	O
out	O
of	O
fashion	O
many	O
times	O
since	O
it	O
was	O
first	O
described	O
almost	O
a	O
century	O
ago	O
.	O

The	O
field	O
is	O
currently	O
in	O
ascendance	O
due	O
to	O
two	O
factors	O
.	O

Over	O
a	O
million	O
FDG	O
-	O
PET	O
studies	O
have	O
unequivocally	O
identified	O
increased	O
glucose	B-Simple_chemical
uptake	O
as	O
a	O
hallmark	O
of	O
metastatic	B-Cancer
cancer	I-Cancer
in	O
humans	B-Organism
.	O

These	O
observations	O
,	O
combined	O
with	O
new	O
molecular	O
insights	O
with	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
and	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
,	O
have	O
rekindled	O
an	O
interest	O
in	O
this	O
important	O
phenotype	O
.	O

A	O
preponderance	O
of	O
work	O
has	O
been	O
focused	O
on	O
the	O
molecular	O
mechanisms	O
underlying	O
this	O
effect	O
,	O
with	O
the	O
expectation	O
that	O
a	O
mechanistic	O
understanding	O
may	O
lead	O
to	O
novel	O
therapeutic	O
approaches	O
.	O

There	O
is	O
also	O
an	O
implicit	O
assumption	O
that	O
a	O
mechanistic	O
understanding	O
,	O
although	O
fundamentally	O
reductionist	O
,	O
will	O
nonetheless	O
lead	O
to	O
a	O
more	O
profound	O
teleological	O
understanding	O
of	O
the	O
need	O
for	O
altered	O
metabolism	O
in	O
invasive	O
cancers	B-Cancer
.	O

In	O
this	O
communication	O
,	O
we	O
describe	O
an	O
alternative	O
approach	O
that	O
begins	O
with	O
teleology	O
;	O
i	O
.	O
e	O
.	O
adaptive	O
landscapes	O
and	O
selection	O
pressures	O
that	O
promote	O
emergence	O
of	O
aerobic	O
glycolysis	O
during	O
the	O
somatic	O
evolution	O
of	O
invasive	O
cancer	B-Cancer
.	O

Mathematical	O
models	O
and	O
empirical	O
observations	O
are	O
used	O
to	O
define	O
the	O
adaptive	O
advantage	O
of	O
aerobic	O
glycolysis	O
that	O
would	O
explain	O
its	O
remarkable	O
prevalence	O
in	O
human	B-Organism
cancers	B-Cancer
.	O

These	O
studies	O
have	O
led	O
to	O
the	O
hypothesis	O
that	O
increased	O
consumption	O
of	O
glucose	B-Simple_chemical
in	O
metastatic	B-Cancer
lesions	I-Cancer
is	O
not	O
used	O
for	O
substantial	O
energy	O
production	O
via	O
Embden	O
-	O
Meyerhoff	O
glycolysis	O
,	O
but	O
rather	O
for	O
production	O
of	O
acid	B-Simple_chemical
,	O
which	O
gives	O
the	O
cancer	B-Cell
cells	I-Cell
a	O
competitive	O
advantage	O
for	O
invasion	O
.	O

Alternative	O
hypotheses	O
,	O
wherein	O
the	O
glucose	B-Simple_chemical
is	O
used	O
for	O
generation	O
of	O
reducing	O
equivalents	O
(	O
NADPH	B-Simple_chemical
)	O
or	O
anabolic	O
precursors	O
(	O
ribose	B-Simple_chemical
)	O
are	O
also	O
discussed	O
.	O

Ectopic	O
decorin	B-Gene_or_gene_product
expression	O
up	O
-	O
regulates	O
VEGF	B-Gene_or_gene_product
expression	O
in	O
mouse	B-Organism
cerebral	B-Cell
endothelial	I-Cell
cells	I-Cell
via	O
activation	O
of	O
the	O
transcription	O
factors	O
Sp1	B-Gene_or_gene_product
,	O
HIF1alpha	B-Gene_or_gene_product
,	O
and	O
Stat3	B-Gene_or_gene_product
.	O

We	O
demonstrate	O
that	O
a	O
proteoglycan	O
decorin	B-Gene_or_gene_product
(	O
DCN	B-Gene_or_gene_product
)	O
up	O
-	O
regulates	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
(	O
VEGF	B-Gene_or_gene_product
)	O
expression	O
with	O
activation	O
of	O
VEGF	B-Gene_or_gene_product
regulating	O
transcription	O
factors	O
Sp1	B-Gene_or_gene_product
,	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
HIF1alpha	B-Gene_or_gene_product
)	O
,	O
and	O
signal	B-Gene_or_gene_product
transducer	I-Gene_or_gene_product
and	I-Gene_or_gene_product
activator	I-Gene_or_gene_product
of	I-Gene_or_gene_product
transcription	I-Gene_or_gene_product
3	I-Gene_or_gene_product
(	O
Stat3	B-Gene_or_gene_product
)	O
via	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
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
extracellular	I-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
/	I-Gene_or_gene_product
2	I-Gene_or_gene_product
(	O
ERK1	B-Gene_or_gene_product
/	I-Gene_or_gene_product
2	I-Gene_or_gene_product
)	O
,	O
and	O
protein	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
B	I-Gene_or_gene_product
(	O
AKT	B-Gene_or_gene_product
)	O
pathways	O
in	O
DCN	B-Gene_or_gene_product
transfected	O
mouse	B-Organism
cerebral	B-Cell
endothelial	I-Cell
(	I-Cell
MCE	I-Cell
)	I-Cell
cells	I-Cell
.	O

Treatment	O
with	O
pharmacological	O
inhibitors	O
and	O
small	O
interfering	O
RNAs	O
reveal	O
that	O
induction	O
and	O
activation	O
of	O
Sp1	B-Gene_or_gene_product
,	O
HIF1alpha	B-Gene_or_gene_product
,	O
and	O
Stat3	B-Gene_or_gene_product
facilitate	O
their	O
nuclear	O
localization	O
and	O
binding	O
to	O
their	O
specific	O
motifs	O
of	O
the	O
VEGF	B-Gene_or_gene_product
promoter	O
and	O
induce	O
VEGF	B-Gene_or_gene_product
expression	O
via	O
two	O
independent	O
pathways	O
,	O
DCN	B-Gene_or_gene_product
/	O
EGFR	B-Gene_or_gene_product
/	O
phosphoinositide	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
/	O
AKT	B-Gene_or_gene_product
and	O
DCN	B-Gene_or_gene_product
/	O
EGFR	B-Gene_or_gene_product
/	O
ERK1	B-Gene_or_gene_product
/	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
respectively	O
,	O
in	O
DCN	B-Gene_or_gene_product
synthesizing	O
MCE	B-Cell
cells	I-Cell
.	O

The	O
cell	B-Cell
type	O
specific	O
glycosylation	O
protects	O
Sp1	B-Gene_or_gene_product
and	O
HIF1alpha	B-Gene_or_gene_product
from	O
proteosome	O
degradation	O
and	O
plays	O
an	O
important	O
and	O
novel	O
role	O
in	O
the	O
regulation	O
of	O
VEGF	B-Gene_or_gene_product
in	O
DCN	B-Gene_or_gene_product
transfected	O
MCE	B-Cell
cells	I-Cell
.	O

Induction	O
of	O
gelatinases	O
(	O
matrix	B-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
9	B-Gene_or_gene_product
)	O
,	O
the	O
serine	O
protease	O
tissue	B-Gene_or_gene_product
plasminogen	I-Gene_or_gene_product
activator	I-Gene_or_gene_product
and	O
plasmin	B-Gene_or_gene_product
by	O
DCN	B-Gene_or_gene_product
transfection	O
in	O
MCE	B-Cell
cells	I-Cell
leads	O
to	O
extracellular	B-Immaterial_anatomical_entity
proteolysis	O
and	O
to	O
release	O
of	O
matrix	O
-	O
bound	O
VEGF	B-Gene_or_gene_product
and	O
activation	O
of	O
angiogenesis	O
.	O

In	O
this	O
study	O
,	O
we	O
demonstrate	O
that	O
two	O
independent	O
downstream	O
signal	O
pathways	O
,	O
DCN	B-Gene_or_gene_product
/	O
EGFR	B-Gene_or_gene_product
/	O
ERK1	B-Gene_or_gene_product
/	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
DCN	B-Gene_or_gene_product
/	O
EGFR	B-Gene_or_gene_product
/	O
phosphoinositide	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
/	O
AKT	B-Gene_or_gene_product
,	O
mediate	O
up	O
-	O
regulation	O
and	O
activation	O
of	O
transcription	O
factors	O
of	O
VEGF	B-Gene_or_gene_product
such	O
as	O
HIF1alpha	B-Gene_or_gene_product
,	O
Stat3	B-Gene_or_gene_product
,	O
and	O
Sp1	B-Gene_or_gene_product
and	O
increase	O
VEGF	B-Gene_or_gene_product
transcription	O
and	O
angiogenesis	O
in	O
MCE	B-Cell
cells	I-Cell
.	O

Effects	O
of	O
spironolactone	B-Simple_chemical
on	O
corneal	B-Multi-tissue_structure
allograft	I-Multi-tissue_structure
survival	O
in	O
the	O
rat	B-Organism
.	O

PURPOSE	O
:	O
Spironolactone	B-Simple_chemical
has	O
recently	O
been	O
shown	O
to	O
have	O
suppressive	O
effects	O
on	O
several	O
immunoactive	O
and	O
proinflammatory	O
cytokines	O
.	O

In	O
this	O
study	O
,	O
we	O
investigated	O
the	O
effects	O
of	O
spironolactone	B-Simple_chemical
on	O
the	O
prevention	O
of	O
corneal	B-Multi-tissue_structure
allograft	I-Multi-tissue_structure
rejection	O
in	O
a	O
MHC	B-Gene_or_gene_product
class	I-Gene_or_gene_product
I	I-Gene_or_gene_product
/	O
II	B-Gene_or_gene_product
mismatch	O
rat	B-Organism
corneal	B-Multi-tissue_structure
transplant	O
model	O
.	O

METHODS	O
:	O
Grafted	O
animals	O
for	O
corneal	B-Multi-tissue_structure
survival	O
analysis	O
were	O
assigned	O
to	O
receive	O
either	O
spironolactone	B-Simple_chemical
suspension	O
(	O
orally	O
,	O
100	O
mg	O
/	O
kg	O
/	O
day	O
,	O
n	O
=	O
7	O
)	O
,	O
phosphate	B-Simple_chemical
-	I-Simple_chemical
buffered	I-Simple_chemical
saline	I-Simple_chemical
(	O
PBS	B-Simple_chemical
,	O
orally	O
,	O
same	O
volume	O
as	O
spironolactone	B-Simple_chemical
,	O
n	O
=	O
9	O
)	O
or	O
remained	O
untreated	O
(	O
n	O
=	O
16	O
)	O
.	O

Additional	O
grafted	O
rats	B-Organism
treated	O
with	O
spironolactone	B-Simple_chemical
(	O
n	O
=	O
6	O
)	O
or	O
PBS	B-Simple_chemical
(	O
n	O
=	O
8	O
)	O
were	O
sacrificed	O
on	O
day	O
12	O
for	O
quantitative	O
RT	O
-	O
PCR	O
analysis	O
for	O
mechanistic	O
studies	O
.	O

RESULTS	O
:	O
Mean	O
(	O
+	O
/	O
-	O
SEM	O
)	O
graft	B-Multi-tissue_structure
survival	O
was	O
significantly	O
prolonged	O
in	O
animals	O
receiving	O
spironolactone	B-Simple_chemical
(	O
14	O
.	O
9	O
+	O
/	O
-	O
2	O
.	O
0	O
days	O
)	O
compared	O
with	O
both	O
PBS	B-Simple_chemical
-	O
treated	O
(	O
12	O
.	O
3	O
+	O
/	O
-	O
1	O
.	O
2	O
days	O
,	O
p	O
=	O
0	O
.	O
007	O
)	O
and	O
untreated	O
controls	O
(	O
13	O
.	O
0	O
+	O
/	O
-	O
1	O
.	O
0	O
days	O
,	O
p	O
=	O
0	O
.	O
01	O
)	O
.	O

We	O
found	O
a	O
decrease	O
in	O
corneal	B-Multi-tissue_structure
neovascularization	O
in	O
spironolactone	B-Simple_chemical
-	O
treated	O
rats	B-Organism
compared	O
with	O
the	O
PBS	B-Simple_chemical
-	O
treated	O
group	O
,	O
although	O
the	O
difference	O
was	O
not	O
statistically	O
significant	O
.	O

Spironolactone	B-Simple_chemical
affected	O
both	O
systemic	O
(	O
down	O
-	O
regulation	O
of	O
CD25	B-Gene_or_gene_product
+	O
cells	O
in	O
spleen	B-Organ
)	O
and	O
local	O
immune	O
response	O
(	O
up	O
-	O
regulation	O
of	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
10	I-Gene_or_gene_product
in	O
cornea	B-Multi-tissue_structure
)	O
.	O

CONCLUSION	O
:	O
We	O
present	O
initial	O
results	O
demonstrating	O
anti	O
-	O
inflammatory	O
effects	O
of	O
spironolactone	B-Simple_chemical
.	O

FAS	B-Gene_or_gene_product
-	O
1377	O
G	O
/	O
A	O
polymorphism	O
and	O
the	O
risk	O
of	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
in	O
cervical	B-Cancer
cancer	I-Cancer
.	O

Single	O
-	O
nucleotide	O
polymorphisms	O
of	O
the	O
FAS	B-Gene_or_gene_product
-	O
1377G	O
/	O
A	O
,	O
FAS	B-Gene_or_gene_product
-	O
670A	O
/	O
G	O
,	O
and	O
FASL	B-Gene_or_gene_product
-	O
844T	O
/	O
C	O
genes	O
may	O
alter	O
transcriptional	O
activity	O
of	O
these	O
genes	O
.	O

Recent	O
evidence	O
suggests	O
an	O
association	O
of	O
these	O
polymorphisms	O
with	O
an	O
increased	O
risk	O
of	O
cervical	B-Cancer
cancer	I-Cancer
,	O
so	O
we	O
explored	O
this	O
relationship	O
.	O

Genotypes	O
of	O
155	O
patients	B-Organism
with	O
cervical	B-Cancer
cancer	I-Cancer
and	O
160	O
healthy	B-Organism
control	I-Organism
subjects	I-Organism
were	O
determined	O
using	O
polymerase	O
chain	O
reaction	O
-	O
based	O
restriction	O
fragment	O
length	O
polymorphism	O
(	O
PCR	O
-	O
RFLP	O
)	O
.	O

Associations	O
with	O
cancer	B-Cancer
risk	O
were	O
estimated	O
using	O
two	O
-	O
sided	O
logistic	O
regression	O
.	O

We	O
observed	O
a	O
significantly	O
increased	O
risk	O
of	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
associated	O
with	O
the	O
FAS	B-Gene_or_gene_product
-	O
1377	O
GA	O
or	O
AA	O
polymorphism	O
[	O
odds	O
ratio	O
(	O
OR	O
)	O
=	O
4	O
.	O
16	O
,	O
95	O
%	O
confidence	O
interval	O
(	O
CI	O
)	O
=	O
1	O
.	O
10	O
to	O
15	O
.	O
74	O
;	O
P	O
=	O
0	O
.	O
036	O
]	O
.	O

In	O
addition	O
,	O
the	O
FAS	B-Gene_or_gene_product
-	O
670AG	O
or	O
GG	O
genotype	O
showed	O
an	O
increased	O
incidence	O
of	O
node	B-Multi-tissue_structure
metastasis	O
,	O
but	O
these	O
findings	O
were	O
not	O
statistically	O
significant	O
(	O
OR	O
=	O
3	O
.	O
67	O
,	O
95	O
%	O
CI	O
=	O
0	O
.	O
96	O
-	O
14	O
.	O
00	O
,	O
P	O
=	O
0	O
.	O
059	O
)	O
.	O

There	O
was	O
no	O
significant	O
association	O
between	O
an	O
increased	O
risk	O
of	O
cervical	B-Cancer
cancer	I-Cancer
and	O
polymorphisms	O
of	O
the	O
death	O
pathway	O
genes	O
FAS	B-Gene_or_gene_product
and	O
FASL	B-Gene_or_gene_product
.	O

None	O
of	O
the	O
polymorphisms	O
were	O
associated	O
with	O
risk	O
of	O
advanced	O
stage	O
or	O
histologic	O
subtype	O
of	O
cervical	B-Cancer
cancer	I-Cancer
.	O

In	O
conclusion	O
,	O
FAS	B-Gene_or_gene_product
-	O
1377	O
G	O
-	O
-	O
>	O
A	O
polymorphism	O
may	O
be	O
associated	O
with	O
an	O
increased	O
risk	O
of	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
in	O
Korean	O
cervical	B-Cancer
cancer	I-Cancer
patients	B-Organism
.	O

Detection	O
of	O
lymphovascular	B-Multi-tissue_structure
invasion	O
in	O
early	O
breast	B-Cancer
cancer	I-Cancer
by	O
D2	B-Gene_or_gene_product
-	I-Gene_or_gene_product
40	I-Gene_or_gene_product
(	O
podoplanin	B-Gene_or_gene_product
)	O
:	O
a	O
clinically	O
useful	O
predictor	O
for	O
axillary	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastases	O
.	O

PURPOSE	O
:	O
The	O
aim	O
of	O
this	O
study	O
was	O
to	O
investigate	O
the	O
use	O
of	O
D2	B-Gene_or_gene_product
-	I-Gene_or_gene_product
40	I-Gene_or_gene_product
for	O
the	O
detection	O
of	O
lymphovascular	B-Multi-tissue_structure
invasion	O
(	O
LVI	O
)	O
in	O
node	O
positive	O
and	O
negative	O
early	O
breast	B-Cancer
cancer	I-Cancer
.	O

LVI	O
is	O
associated	O
with	O
axillary	B-Multi-tissue_structure
lymph	I-Multi-tissue_structure
node	I-Multi-tissue_structure
metastases	O
(	O
ALNM	O
)	O
and	O
a	O
long	O
-	O
term	O
prognostic	O
factor	O
.	O

A	O
precise	O
identification	O
of	O
LVI	O
would	O
have	O
a	O
strong	O
clinical	O
impact	O
for	O
breast	B-Cancer
cancer	I-Cancer
patients	B-Organism
.	O

METHODS	O
:	O
Immunohistochemical	O
staining	O
with	O
D2	B-Gene_or_gene_product
-	I-Gene_or_gene_product
40	I-Gene_or_gene_product
and	O
CD34	B-Gene_or_gene_product
was	O
performed	O
on	O
formalin	O
-	O
fixed	O
,	O
paraffin	O
-	O
embedded	O
tissue	B-Tissue
sections	I-Tissue
of	O
254	O
invasive	O
breast	B-Cancer
tumors	I-Cancer
of	O
247	O
patients	B-Organism
with	O
node	O
negative	O
and	O
node	O
positive	O
early	O
breast	B-Cancer
cancer	I-Cancer
.	O

All	O
slides	O
were	O
screened	O
for	O
the	O
presence	O
of	O
LVI	O
.	O

Correlation	O
with	O
clinico	O
-	O
pathological	O
factors	O
including	O
LVI	O
as	O
retrieved	O
by	O
routine	O
haematoxylin	B-Simple_chemical
and	O
eosin	B-Simple_chemical
(	O
H	B-Simple_chemical
.	I-Simple_chemical
E	I-Simple_chemical
.	I-Simple_chemical
)	O
stained	O
sections	B-Tissue
and	O
the	O
eligibility	O
for	O
the	O
prediction	O
of	O
ALNM	O
was	O
assessed	O
.	O

RESULTS	O
:	O
Using	O
the	O
D2	B-Gene_or_gene_product
-	I-Gene_or_gene_product
40	I-Gene_or_gene_product
antibody	O
for	O
immunostaining	O
,	O
our	O
results	O
demonstrate	O
a	O
significant	O
higher	O
detection	O
(	O
P	O
<	O
0	O
.	O
001	O
)	O
of	O
LVI	O
as	O
compared	O
with	O
routine	O
H	B-Simple_chemical
.	I-Simple_chemical
E	I-Simple_chemical
.	I-Simple_chemical
-	O
staining	O
in	O
early	O
breast	B-Cancer
cancer	I-Cancer
.	O

LVI	O
was	O
correctly	O
identified	O
by	O
D2	B-Gene_or_gene_product
-	I-Gene_or_gene_product
40	I-Gene_or_gene_product
(	O
D2	B-Gene_or_gene_product
-	I-Gene_or_gene_product
40	I-Gene_or_gene_product
+	O
)	O
in	O
70	O
out	O
of	O
254	O
tumors	B-Cancer
(	O
28	O
%	O
)	O
as	O
compared	O
to	O
40	O
tumors	B-Cancer
(	O
16	O
%	O
)	O
by	O
routine	O
HE	B-Simple_chemical
staining	O
(	O
HE	B-Simple_chemical
+	O
)	O
.	O

There	O
was	O
a	O
significant	O
correlation	O
between	O
D2	B-Gene_or_gene_product
-	I-Gene_or_gene_product
40	I-Gene_or_gene_product
+	O
LVI	O
and	O
age	O
,	O
t	O
-	O
stage	O
,	O
nodal	O
status	O
,	O
grading	O
and	O
hormonreceptor	O
-	O
status	O
.	O

Correlation	O
between	O
D2	B-Gene_or_gene_product
-	I-Gene_or_gene_product
40	I-Gene_or_gene_product
+	O
LVI	O
and	O
menopausal	O
-	O
status	O
,	O
HER2	B-Gene_or_gene_product
-	O
status	O
and	O
histological	O
type	O
was	O
not	O
significant	O
,	O
while	O
there	O
was	O
a	O
significant	O
correlation	O
of	O
D2	B-Gene_or_gene_product
-	I-Gene_or_gene_product
40	I-Gene_or_gene_product
and	O
so	O
called	O
"	O
triple	O
negative	O
"	O
tumors	B-Cancer
(	O
ER	B-Gene_or_gene_product
/	O
PR	B-Gene_or_gene_product
and	O
HER2neu	B-Gene_or_gene_product
-	O
negative	O
)	O
.	O

In	O
a	O
multivariate	O
analysis	O
D2	B-Gene_or_gene_product
-	I-Gene_or_gene_product
40	I-Gene_or_gene_product
+	O
was	O
the	O
strongest	O
predictor	O
for	O
ALNM	O
with	O
an	O
odds	O
ratio	O
of	O
3	O
.	O
489	O
and	O
a	O
P	O
-	O
value	O
of	O
P	O
=	O
0	O
.	O
0003	O
,	O
followed	O
only	O
by	O
T	O
-	O
stage	O
and	O
grading	O
with	O
odds	O
ratios	O
of	O
3	O
.	O
167	O
and	O
1	O
.	O
953	O
and	O
P	O
-	O
values	O
P	O
=	O
0	O
.	O
0003	O
and	O
P	O
=	O
0	O
.	O
0352	O
.	O

CONCLUSION	O
:	O
Immunostaining	O
with	O
D2	B-Gene_or_gene_product
-	I-Gene_or_gene_product
40	I-Gene_or_gene_product
significantly	O
increased	O
the	O
frequency	O
of	O
detection	O
of	O
lymphatic	B-Multi-tissue_structure
invasion	O
compared	O
to	O
conventional	O
H	B-Simple_chemical
.	I-Simple_chemical
E	I-Simple_chemical
.	I-Simple_chemical
-	O
staining	O
in	O
early	O
breast	B-Cancer
cancer	I-Cancer
.	O

As	O
LVI	O
is	O
a	O
strong	O
predictive	O
and	O
prognostic	O
marker	O
,	O
the	O
monoclonal	O
antibody	O
D2	B-Gene_or_gene_product
-	I-Gene_or_gene_product
40	I-Gene_or_gene_product
has	O
the	O
potential	O
to	O
play	O
a	O
significant	O
role	O
in	O
pathological	O
routine	O
workup	O
of	O
breast	B-Cancer
tumors	I-Cancer
.	O

Further	O
prospective	O
studies	O
are	O
needed	O
to	O
prove	O
the	O
clinical	O
impact	O
of	O
D2	B-Gene_or_gene_product
-	I-Gene_or_gene_product
40	I-Gene_or_gene_product
.	O

N	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
augments	O
death	O
and	O
attenuates	O
protective	O
effects	O
of	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
in	O
trophically	O
stressed	O
neuroblastoma	B-Cell
cells	I-Cell
.	O

N	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
has	O
proapoptotic	O
functions	O
,	O
yet	O
it	O
acts	O
as	O
an	O
oncogene	O
in	O
neuroblastoma	B-Cancer
.	O

Thus	O
,	O
antiapoptotic	O
mechanisms	O
have	O
to	O
be	O
operative	O
in	O
neuroblastoma	B-Cell
cells	I-Cell
that	O
antagonize	O
the	O
proapoptotic	O
effects	O
of	O
N	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
.	O

We	O
conditionally	O
activated	O
N	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
in	O
SH	B-Cell
-	I-Cell
EP	I-Cell
neuroblastoma	I-Cell
cells	I-Cell
subjected	O
to	O
the	O
trophic	O
stress	O
of	O
serum	B-Organism_substance
or	O
nutrient	O
deprivation	O
while	O
changing	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
survivin	B-Gene_or_gene_product
and	O
FLIP	B-Gene_or_gene_product
(	I-Gene_or_gene_product
L	I-Gene_or_gene_product
)	I-Gene_or_gene_product
,	O
antiapoptotic	O
molecules	O
often	O
overexpressed	O
in	O
poor	O
prognosis	O
neuroblastomas	B-Cancer
.	O

Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
protected	O
SH	B-Cell
-	I-Cell
EP	I-Cell
cells	I-Cell
from	O
death	O
during	O
nutritional	O
deprivation	O
by	O
activating	O
energetically	O
advantageous	O
oxidative	O
phosphorylation	O
.	O

N	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
overrode	O
the	O
metabolic	O
protection	O
provided	O
by	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
-	O
induced	O
oxidative	O
phosphorylation	O
by	O
reestablishing	O
the	O
glycolytic	O
phenotype	O
and	O
attenuated	O
the	O
antiapoptotic	O
effect	O
of	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
during	O
metabolic	O
stress	O
.	O

Survivin	B-Gene_or_gene_product
partially	O
antagonized	O
the	O
growth	O
suppressive	O
function	O
of	O
N	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
in	O
SH	B-Cell
-	I-Cell
EP	I-Cell
neuroblastoma	I-Cell
cells	I-Cell
during	O
serum	B-Organism_substance
deprivation	O
whereas	O
FLIP	B-Gene_or_gene_product
(	I-Gene_or_gene_product
L	I-Gene_or_gene_product
)	I-Gene_or_gene_product
did	O
not	O
.	O

These	O
findings	O
advance	O
our	O
understanding	O
of	O
the	O
functions	O
of	O
N	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
in	O
neuroblastoma	B-Cell
cells	I-Cell
.	O

The	O
tyrosine	O
kinase	O
inhibitor	O
cediranib	B-Simple_chemical
blocks	O
ligand	O
-	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
receptor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
activity	O
and	O
lymphangiogenesis	O
.	O

Solid	B-Cancer
tumors	I-Cancer
express	O
a	O
range	O
of	O
factors	O
required	O
to	O
sustain	O
their	O
growth	O
and	O
promote	O
their	O
dissemination	O
.	O

Among	O
these	O
are	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
A	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
A	I-Gene_or_gene_product
)	O
,	O
the	O
key	O
angiogenic	O
stimulant	O
,	O
and	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
,	O
a	O
primary	O
mediator	O
of	O
lymphangiogenesis	O
.	O

Small	O
molecule	O
tyrosine	B-Simple_chemical
kinase	I-Simple_chemical
inhibitors	I-Simple_chemical
offer	O
the	O
potential	O
to	O
inhibit	O
more	O
than	O
one	O
kinase	O
and	O
impede	O
tumor	B-Cancer
growth	O
by	O
multiple	O
mechanisms	O
.	O

However	O
,	O
their	O
potency	O
toward	O
individual	O
targets	O
can	O
vary	O
.	O

Cediranib	B-Simple_chemical
(	O
RECENTIN	B-Simple_chemical
;	O
AZD2171	B-Simple_chemical
)	O
is	O
an	O
inhibitor	O
of	O
VEGF	B-Gene_or_gene_product
signaling	O
that	O
has	O
been	O
shown	O
in	O
experimental	O
models	O
to	O
prevent	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
A	I-Gene_or_gene_product
-	O
induced	O
angiogenesis	O
and	O
primary	O
tumor	B-Cancer
growth	O
,	O
yet	O
the	O
effects	O
of	O
cediranib	B-Simple_chemical
on	O
VEGF	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
(	I-Gene_or_gene_product
VEGFR	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
-	O
mediated	O
endothelial	B-Cell
cell	I-Cell
function	O
and	O
lymphangiogenesis	O
are	O
unknown	O
.	O

To	O
better	O
understand	O
the	O
activity	O
of	O
cediranib	B-Simple_chemical
against	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
and	O
its	O
associated	O
signaling	O
events	O
compared	O
with	O
its	O
activity	O
against	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
we	O
used	O
the	O
receptor	O
-	O
specific	O
ligands	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
E	I-Gene_or_gene_product
and	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C156S	I-Gene_or_gene_product
.	O

In	O
human	B-Organism
endothelial	B-Cell
cells	I-Cell
,	O
cediranib	B-Simple_chemical
inhibited	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
E	I-Gene_or_gene_product
-	O
induced	O
phosphorylation	O
of	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C156S	I-Gene_or_gene_product
-	O
induced	O
phosphorylation	O
of	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
at	O
concentrations	O
of	O
less	O
than	O
/	O
=	O
1nmol	O
/	O
L	O
and	O
inhibited	O
activation	O
of	O
downstream	O
signaling	O
molecules	O
.	O

Additionally	O
,	O
cediranib	B-Simple_chemical
blocked	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C156S	I-Gene_or_gene_product
-	O
induced	O
and	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
E	I-Gene_or_gene_product
-	O
induced	O
proliferation	O
,	O
survival	O
,	O
and	O
migration	O
of	O
lymphatic	B-Cell
and	O
blood	B-Cell
vascular	I-Cell
endothelial	I-Cell
cells	I-Cell
.	O

In	O
vivo	O
,	O
cediranib	B-Simple_chemical
(	O
6	O
mg	O
/	O
kg	O
/	O
d	O
)	O
prevented	O
angiogenesis	O
and	O
lymphangiogenesis	O
induced	O
by	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
E	I-Gene_or_gene_product
-	O
expressing	O
and	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C156S	I-Gene_or_gene_product
-	O
expressing	O
adenoviruses	B-Organism
,	O
respectively	O
.	O

Cediranib	B-Simple_chemical
(	O
6	O
mg	O
/	O
kg	O
/	O
day	O
)	O
also	O
blocked	O
angiogenesis	O
and	O
lymphangiogenesis	O
induced	O
by	O
adenoviruses	B-Organism
expressing	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
A	I-Gene_or_gene_product
or	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
and	O
compromised	O
the	O
blood	B-Organism_substance
and	O
lymphatic	B-Multi-tissue_structure
vasculatures	I-Multi-tissue_structure
of	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
-	O
expressing	O
tumors	B-Cancer
.	O

Cediranib	B-Simple_chemical
may	O
,	O
therefore	O
,	O
be	O
an	O
effective	O
means	O
of	O
preventing	O
tumor	B-Cancer
progression	O
,	O
not	O
only	O
by	O
inhibiting	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
activity	O
and	O
angiogenesis	O
,	O
but	O
also	O
by	O
concomitantly	O
inhibiting	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
activity	O
and	O
lymphangiogenesis	O
.	O

Artemisinin	B-Simple_chemical
inhibits	O
tumor	B-Cancer
lymphangiogenesis	O
by	O
suppression	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
C	I-Gene_or_gene_product
.	O

We	O
have	O
previously	O
reported	O
that	O
dihydroartemisinin	B-Simple_chemical
is	O
found	O
to	O
have	O
a	O
potent	O
ability	O
in	O
influencing	O
lymphatic	B-Cell
endothelial	I-Cell
cell	I-Cell
migration	O
and	O
tube	B-Tissue
formation	O
.	O

In	O
this	O
study	O
,	O
we	O
investigated	O
the	O
effect	O
of	O
artemisinin	B-Simple_chemical
on	O
tumor	B-Cancer
growth	O
,	O
lymphangiogenesis	O
,	O
metastasis	O
and	O
survival	O
in	O
mouse	B-Organism
Lewis	B-Cancer
lung	I-Cancer
carcinoma	I-Cancer
(	O
LLC	B-Cancer
)	O
models	O
.	O

We	O
found	O
that	O
orally	B-Organism_subdivision
administered	O
artemisinin	B-Simple_chemical
inhibited	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
and	O
lung	B-Organ
metastasis	O
and	O
prolonged	O
survival	O
without	O
retarding	O
tumor	B-Cancer
growth	O
.	O

Consistent	O
with	O
the	O
decrease	O
in	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
,	O
tumor	B-Cancer
lymphangiogenesis	O
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
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
was	O
significantly	O
decreased	O
in	O
artemisinin	B-Simple_chemical
-	O
treated	O
mice	B-Organism
,	O
as	O
compared	O
to	O
control	O
mice	B-Organism
.	O

Furthermore	O
,	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1beta	I-Gene_or_gene_product
-	O
induced	O
p38	B-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
activation	O
and	O
upregulation	O
of	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
mRNA	O
and	O
protein	O
in	O
LLC	B-Cell
cells	I-Cell
was	O
also	O
suppressed	O
by	O
artemisinin	B-Simple_chemical
or	O
by	O
the	O
p38	B-Gene_or_gene_product
MAPK	I-Gene_or_gene_product
inhibitor	O
SB	B-Simple_chemical
-	I-Simple_chemical
203580	I-Simple_chemical
,	O
suggesting	O
that	O
p38	B-Gene_or_gene_product
MAPK	I-Gene_or_gene_product
could	O
serve	O
as	O
a	O
mediator	O
of	O
proinflammatory	O
cytokine	O
-	O
induced	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
expression	O
.	O

These	O
data	O
indicate	O
that	O
artemisinin	B-Simple_chemical
may	O
be	O
useful	O
for	O
the	O
prevention	O
of	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
by	O
downregulating	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
C	I-Gene_or_gene_product
and	O
reducing	O
tumor	B-Cancer
lymphangiogenesis	O
.	O

Bevacizumab	B-Simple_chemical
and	O
irinotecan	B-Simple_chemical
therapy	O
in	O
glioblastoma	B-Cancer
multiforme	I-Cancer
:	O
a	O
series	O
of	O
13	O
cases	O
.	O

OBJECT	O
:	O
Endothelial	B-Cell
proliferation	O
has	O
been	O
recognized	O
as	O
a	O
marker	O
of	O
high	O
-	O
grade	O
or	O
aggressive	O
glioma	B-Cancer
.	O

Bevacizumab	B-Simple_chemical
is	O
a	O
humanized	O
immunoglobulin	B-Gene_or_gene_product
G1	I-Gene_or_gene_product
monoclonal	B-Gene_or_gene_product
antibody	I-Gene_or_gene_product
to	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
that	O
has	O
been	O
shown	O
to	O
have	O
activity	O
in	O
malignant	B-Cancer
gliomas	I-Cancer
when	O
combined	O
with	O
irinotecan	B-Simple_chemical
.	O

The	O
authors	O
report	O
on	O
a	O
case	O
series	O
of	O
13	O
patients	B-Organism
with	O
recurrent	O
heavily	O
pretreated	O
malignant	B-Cancer
glioma	I-Cancer
that	O
was	O
treated	O
with	O
the	O
combination	O
of	O
bevacizumab	B-Simple_chemical
and	O
irinotecan	B-Simple_chemical
.	O

METHODS	O
:	O
Standard	O
therapy	O
with	O
primary	O
resection	O
followed	O
by	O
adjuvant	O
chemotherapy	O
and	O
radiation	O
had	O
failed	O
in	O
all	O
patients	B-Organism
.	O

The	O
median	O
number	O
of	O
therapies	O
applied	O
,	O
including	O
initial	O
surgery	O
,	O
was	O
5	O
(	O
range	O
3	O
-	O
7	O
therapies	O
)	O
.	O

Nine	O
patients	B-Organism
were	O
started	O
on	O
bevacizumab	B-Simple_chemical
at	O
a	O
dose	O
of	O
5	O
mg	O
/	O
m2	O
every	O
2	O
weeks	O
.	O

Four	O
patients	B-Organism
received	O
bevacizumab	B-Simple_chemical
at	O
a	O
dose	O
of	O
10	O
mg	O
/	O
m2	O
;	O
irinotecan	B-Simple_chemical
was	O
given	O
at	O
a	O
dose	O
of	O
125	O
mg	O
/	O
m2	O
every	O
week	O
for	O
3	O
weeks	O
.	O

RESULTS	O
:	O
Of	O
the	O
13	O
treated	O
patients	B-Organism
,	O
10	O
(	O
77	O
%	O
)	O
had	O
a	O
radiologically	O
demonstrated	O
partial	O
response	O
and	O
3	O
(	O
23	O
%	O
)	O
had	O
stable	O
disease	O
.	O

Six	O
patients	B-Organism
(	O
46	O
%	O
)	O
had	O
a	O
clinical	O
response	O
.	O

The	O
median	O
time	O
to	O
disease	O
progression	O
while	O
on	O
treatment	O
was	O
24	O
weeks	O
.	O

The	O
median	O
overall	O
survival	O
was	O
27	O
weeks	O
.	O

The	O
disease	O
progressed	O
in	O
8	O
patients	B-Organism
,	O
despite	O
an	O
initial	O
response	O
.	O

Five	O
patients	B-Organism
are	O
still	O
responding	O
to	O
therapy	O
.	O

Six	O
of	O
the	O
8	O
patients	B-Organism
whose	O
disease	O
progressed	O
have	O
died	O
.	O

Bevacizumab	B-Simple_chemical
was	O
discontinued	O
in	O
2	O
patients	B-Organism
because	O
of	O
nonfatal	O
intracranial	B-Immaterial_anatomical_entity
bleeding	O
.	O

CONCLUSIONS	O
:	O
The	O
combination	O
of	O
bevacizumab	B-Simple_chemical
and	O
irinotecan	B-Simple_chemical
is	O
safe	O
and	O
has	O
excellent	O
activity	O
even	O
in	O
this	O
relapsed	O
,	O
heavily	O
pretreated	O
population	O
of	O
patients	B-Organism
with	O
high	O
-	O
grade	O
malignant	B-Cancer
glioma	I-Cancer
,	O
most	O
of	O
whom	O
would	O
not	O
be	O
candidates	O
for	O
clinical	O
trials	O
.	O

Endosialin	B-Gene_or_gene_product
/	O
TEM	B-Gene_or_gene_product
1	I-Gene_or_gene_product
/	O
CD248	B-Gene_or_gene_product
is	O
a	O
pericyte	B-Cell
marker	O
of	O
embryonic	B-Developing_anatomical_structure
and	O
tumor	B-Cancer
neovascularization	O
.	O

The	O
formation	O
of	O
functional	O
,	O
mature	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
depends	O
on	O
the	O
interaction	O
between	O
endothelial	B-Cell
cells	I-Cell
and	O
pericytes	B-Cell
.	O

Commonality	O
exists	O
in	O
the	O
processes	O
involved	O
in	O
vasculature	B-Multi-tissue_structure
development	O
between	O
tissues	B-Tissue
whether	O
healthy	O
or	O
diseased	O
.	O

Endosialin	B-Gene_or_gene_product
/	O
TEM	B-Gene_or_gene_product
1	I-Gene_or_gene_product
is	O
a	O
cell	B-Cellular_component
membrane	I-Cellular_component
protein	O
that	O
is	O
expressed	O
in	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
during	O
embryogenesis	O
and	O
tumorigenesis	O
but	O
not	O
in	O
normal	O
mature	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
.	O

Antibodies	O
developed	O
to	O
human	B-Organism
endosialin	B-Gene_or_gene_product
were	O
used	O
to	O
investigate	O
endosialin	B-Gene_or_gene_product
expression	O
and	O
function	O
in	O
human	B-Organism
prenatal	B-Cell
brain	I-Cell
pericytes	I-Cell
and	O
pericytes	B-Cell
residing	O
in	O
tumors	B-Cancer
.	O

Anti	B-Gene_or_gene_product
-	I-Gene_or_gene_product
endosialin	I-Gene_or_gene_product
was	O
capable	O
of	O
preventing	O
pericyte	B-Cell
tube	B-Tissue
formation	O
in	O
culture	B-Cell
and	O
inhibited	O
migration	O
.	O

Brain	B-Cell
pericytes	I-Cell
in	O
culture	O
had	O
higher	O
levels	O
of	O
endosialin	B-Gene_or_gene_product
/	O
TEM	B-Gene_or_gene_product
1	I-Gene_or_gene_product
than	O
TEMs	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
-	B-Gene_or_gene_product
3	I-Gene_or_gene_product
,	O
-	B-Gene_or_gene_product
4	I-Gene_or_gene_product
,	O
-	B-Gene_or_gene_product
5	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
8	I-Gene_or_gene_product
.	O

Immunocytochemistry	O
revealed	O
that	O
endosialin	B-Gene_or_gene_product
was	O
present	O
in	O
the	O
cytoplasmic	B-Organism_substance
body	I-Organism_substance
and	O
in	O
the	O
elongated	O
extensions	O
essential	O
to	O
pericyte	B-Cell
function	O
.	O

Transgenic	O
mice	B-Organism
engineered	O
to	O
express	O
human	B-Organism
endosialin	B-Gene_or_gene_product
bred	O
on	O
an	O
immunocompromised	O
background	O
allowed	O
the	O
growth	O
of	O
human	B-Organism
tumor	B-Cancer
xenografts	I-Cancer
.	O

In	O
human	B-Organism
colon	B-Cancer
carcinoma	I-Cancer
Colo205	I-Cancer
and	O
HT29	B-Cancer
xenografts	I-Cancer
grown	O
in	O
human	B-Organism
endosialin	B-Gene_or_gene_product
-	O
transgenic	O
mice	B-Organism
,	O
endosialin	B-Gene_or_gene_product
expression	O
was	O
largely	O
confined	O
to	O
NG2	B-Gene_or_gene_product
-	O
expressing	O
perivascular	B-Cell
cells	I-Cell
and	O
not	O
CD31	B-Gene_or_gene_product
-	O
positive	O
endothelial	B-Cell
cells	I-Cell
.	O

Similar	O
methods	O
applied	O
to	O
human	B-Organism
ovarian	B-Cancer
and	O
colon	B-Cancer
tumors	I-Cancer
confirmed	O
endosialin	B-Gene_or_gene_product
expression	O
by	O
pericytes	B-Cell
.	O

The	O
data	O
indicate	O
that	O
endosialin	B-Gene_or_gene_product
is	O
strongly	O
expressed	O
by	O
pericytes	B-Cell
during	O
periods	O
of	O
active	O
angiogenesis	O
during	O
embryonic	B-Developing_anatomical_structure
and	O
tumor	B-Cancer
development	O
.	O

Anti	B-Gene_or_gene_product
-	I-Gene_or_gene_product
endosialin	I-Gene_or_gene_product
antibodies	I-Gene_or_gene_product
may	O
have	O
value	O
in	O
identifying	O
vasculature	B-Multi-tissue_structure
in	O
malignant	B-Tissue
tissues	I-Tissue
.	O

With	O
the	O
appropriate	O
agent	O
,	O
targeting	O
endosialin	B-Gene_or_gene_product
may	O
interfere	O
with	O
blood	B-Multi-tissue_structure
vessel	I-Multi-tissue_structure
growth	O
during	O
tumor	B-Cancer
development	O
.	O

Efficient	O
inhibition	O
of	O
ovarian	B-Cancer
cancer	I-Cancer
growth	O
and	O
prolonged	O
survival	O
by	O
transfection	O
with	O
a	O
novel	O
pro	O
-	O
apoptotic	O
gene	O
,	O
hPNAS	B-Gene_or_gene_product
-	I-Gene_or_gene_product
4	I-Gene_or_gene_product
,	O
in	O
a	O
mouse	B-Organism
model	O
.	O

In	O
vivo	O
and	O
in	O
vitro	O
results	O
.	O

OBJECTIVE	O
:	O
We	O
transfected	O
ovarian	B-Cell
cancer	I-Cell
cells	I-Cell
and	O
administered	O
recombinant	O
plasmid	O
encoding	O
hPNAS	B-Gene_or_gene_product
-	I-Gene_or_gene_product
4	I-Gene_or_gene_product
to	O
nude	B-Organism
mice	I-Organism
bearing	O
ovarian	B-Cancer
cancer	I-Cancer
,	O
aiming	O
to	O
evaluate	O
the	O
effect	O
of	O
hPNAS	B-Gene_or_gene_product
-	I-Gene_or_gene_product
4	I-Gene_or_gene_product
against	O
ovarian	B-Cancer
cancer	I-Cancer
in	O
vitro	O
and	O
in	O
vivo	O
.	O

METHODS	O
:	O
Ovarian	B-Cell
cancer	I-Cell
SKOV3	I-Cell
cells	I-Cell
were	O
transfected	O
with	O
hPNAS	B-Gene_or_gene_product
-	I-Gene_or_gene_product
4	I-Gene_or_gene_product
-	O
plasmid	O
,	O
and	O
cell	B-Cell
proliferation	O
was	O
evaluated	O
by	O
MTT	O
assay	O
;	O
apoptosis	O
was	O
examined	O
by	O
DNA	B-Cellular_component
ladder	O
,	O
Hoechst33258	O
staining	O
and	O
flow	O
-	O
cytometric	O
assays	O
.	O

Nude	B-Organism
mice	I-Organism
bearing	O
ovarian	B-Cancer
cancers	I-Cancer
were	O
treated	O
with	O
hPNAS	B-Gene_or_gene_product
-	I-Gene_or_gene_product
4	I-Gene_or_gene_product
-	O
p	O
/	O
liposome	B-Simple_chemical
.	O

Tumor	B-Cancer
growth	O
was	O
determined	O
and	O
survival	O
was	O
recorded	O
.	O

TUNEL	O
assay	O
and	O
microvessel	B-Tissue
density	O
was	O
assessed	O
to	O
evaluate	O
apoptosis	O
and	O
angiogenesis	O
.	O

RESULTS	O
:	O
Both	O
inhibition	O
of	O
proliferation	O
(	O
p	O
<	O
0	O
.	O
05	O
)	O
and	O
induction	O
of	O
apoptosis	O
(	O
p	O
<	O
0	O
.	O
05	O
)	O
were	O
observed	O
in	O
SKOV3	B-Cell
cells	I-Cell
transfected	O
with	O
hPNAS	B-Gene_or_gene_product
-	I-Gene_or_gene_product
4	I-Gene_or_gene_product
-	O
p	O
in	O
vitro	O
.	O

In	O
hPNAS	B-Gene_or_gene_product
-	I-Gene_or_gene_product
4	I-Gene_or_gene_product
-	O
p	O
-	O
treated	O
tumor	B-Cell
cells	I-Cell
in	O
vivo	O
,	O
tumor	B-Cancer
growth	O
significantly	O
decreased	O
,	O
while	O
the	O
survival	O
time	O
of	O
tumor	B-Cancer
-	O
bearing	O
mice	B-Organism
was	O
prolonged	O
compared	O
with	O
control	O
groups	O
(	O
p	O
<	O
0	O
.	O
05	O
)	O
.	O

Increased	O
apoptosis	O
of	O
tumor	B-Cell
cells	I-Cell
and	O
decreased	O
angiogenesis	O
in	O
tumor	B-Tissue
tissue	I-Tissue
were	O
also	O
observed	O
.	O

CONCLUSIONS	O
:	O
Our	O
promising	O
results	O
on	O
the	O
potential	O
antitumor	B-Cancer
effects	O
of	O
hPNAS	B-Gene_or_gene_product
-	I-Gene_or_gene_product
4	I-Gene_or_gene_product
on	O
ovarian	B-Cancer
cancer	I-Cancer
in	O
vitro	O
and	O
in	O
vivo	O
may	O
be	O
explained	O
,	O
in	O
part	O
,	O
by	O
the	O
induction	O
of	O
apoptosis	O
and	O
inhibition	O
of	O
angiogenesis	O
.	O

Consequently	O
,	O
hPNAS	B-Gene_or_gene_product
-	I-Gene_or_gene_product
4	I-Gene_or_gene_product
has	O
potential	O
as	O
a	O
new	O
gene	O
therapy	O
for	O
human	B-Organism
ovarian	B-Cancer
cancer	I-Cancer
.	O

The	O
in	O
vivo	O
properties	O
of	O
STX243	B-Simple_chemical
:	O
a	O
potent	O
angiogenesis	O
inhibitor	O
in	O
breast	B-Cancer
cancer	I-Cancer
.	O

The	O
steroidal	O
-	O
based	O
drug	O
2	B-Simple_chemical
-	I-Simple_chemical
ethyloestradiol	I-Simple_chemical
-	I-Simple_chemical
3	I-Simple_chemical
,	I-Simple_chemical
17	I-Simple_chemical
-	I-Simple_chemical
O	I-Simple_chemical
,	I-Simple_chemical
O	I-Simple_chemical
-	I-Simple_chemical
bis	I-Simple_chemical
-	I-Simple_chemical
sulphamate	I-Simple_chemical
(	O
STX243	B-Simple_chemical
)	O
has	O
been	O
developed	O
as	O
a	O
potent	O
antiangiogenic	O
and	O
antitumour	B-Cancer
compound	O
.	O

The	O
objective	O
of	O
this	O
study	O
was	O
to	O
ascertain	O
whether	O
STX243	B-Simple_chemical
is	O
more	O
active	O
in	O
vivo	O
than	O
the	O
clinically	O
relevant	O
drug	O
2	B-Simple_chemical
-	I-Simple_chemical
methoxyoestradiol	I-Simple_chemical
(	O
2	B-Simple_chemical
-	I-Simple_chemical
MeOE2	I-Simple_chemical
)	O
and	O
the	O
structurally	O
similar	O
compound	O
2	B-Simple_chemical
-	I-Simple_chemical
MeOE2	I-Simple_chemical
-	I-Simple_chemical
3	I-Simple_chemical
,	I-Simple_chemical
17	I-Simple_chemical
-	I-Simple_chemical
O	I-Simple_chemical
,	I-Simple_chemical
O	I-Simple_chemical
-	I-Simple_chemical
bis	I-Simple_chemical
-	I-Simple_chemical
sulphamate	I-Simple_chemical
(	O
STX140	B-Simple_chemical
)	O
.	O

The	O
tumour	B-Cancer
growth	O
inhibition	O
efficacy	O
,	O
antiangiogenic	O
potential	O
and	O
pharmacokinetics	O
of	O
STX243	B-Simple_chemical
were	O
examined	O
using	O
four	O
in	O
vivo	O
models	O
.	O

Both	O
STX243	B-Simple_chemical
and	O
STX140	B-Simple_chemical
were	O
capable	O
of	O
retarding	O
the	O
growth	O
of	O
MDA	B-Cancer
-	I-Cancer
MB	I-Cancer
-	I-Cancer
231	I-Cancer
xenograft	I-Cancer
tumours	I-Cancer
(	O
72	O
and	O
63	O
%	O
,	O
respectively	O
)	O
,	O
whereas	O
no	O
inhibition	O
was	O
observed	O
for	O
animals	O
treated	O
with	O
2	B-Simple_chemical
-	I-Simple_chemical
MeOE2	I-Simple_chemical
.	O

Further	O
tumour	B-Cancer
inhibition	O
studies	O
showed	O
that	O
STX243	B-Simple_chemical
was	O
also	O
active	O
against	O
MCF	O
-	O
7	O
paclitaxel	B-Simple_chemical
-	O
resistant	O
tumours	O
.	O

Using	O
a	O
Matrigel	O
plug	O
-	O
based	O
model	O
,	O
in	O
vivo	O
angiogenesis	O
was	O
restricted	O
with	O
STX243	B-Simple_chemical
and	O
STX140	B-Simple_chemical
(	O
50	O
and	O
72	O
%	O
,	O
respectively	O
,	O
using	O
a	O
10	O
mg	O
kg	O
(	O
-	O
1	O
)	O
oral	O
dose	O
)	O
,	O
thereby	O
showing	O
the	O
antiangiogenic	O
activity	O
of	O
both	O
compounds	O
.	O

The	O
pharmacokinetics	O
of	O
STX243	B-Simple_chemical
were	O
examined	O
at	O
two	O
different	O
doses	O
using	O
adult	O
female	O
rats	B-Organism
.	O

The	O
compound	O
was	O
orally	O
bioavailable	O
(	O
31	O
%	O
after	O
a	O
single	O
10	O
mg	O
kg	O
(	O
-	O
1	O
)	O
dose	O
)	O
and	O
resistant	O
to	O
metabolism	O
.	O

These	O
results	O
show	O
that	O
STX243	B-Simple_chemical
is	O
a	O
potent	O
in	O
vivo	O
drug	O
and	O
could	O
be	O
clinically	O
effective	O
at	O
treating	O
a	O
number	O
of	O
oncological	O
conditions	O
.	O

Grape	B-Organism_substance
seed	I-Organism_substance
extract	I-Organism_substance
inhibits	O
angiogenesis	O
via	O
suppression	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
signaling	O
pathway	O
.	O

Blockade	O
of	O
angiogenesis	O
is	O
an	O
important	O
approach	O
for	O
cancer	B-Cancer
treatment	O
and	O
prevention	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
one	O
of	O
the	O
most	O
critical	O
factors	O
that	O
induce	O
angiogenesis	O
and	O
has	O
thus	O
become	O
an	O
attractive	O
target	O
for	O
antiangiogenesis	O
treatment	O
.	O

However	O
,	O
most	O
current	O
anti	O
-	O
VEGF	B-Gene_or_gene_product
agents	O
often	O
cause	O
some	O
side	O
effects	O
when	O
given	O
chronically	O
.	O

Identification	O
of	O
naturally	O
occurring	O
VEGF	B-Gene_or_gene_product
inhibitors	O
derived	O
from	O
diet	O
would	O
be	O
one	O
alternative	O
approach	O
with	O
an	O
advantage	O
of	O
known	O
safety	O
.	O

Grape	B-Organism_substance
seed	I-Organism_substance
extract	I-Organism_substance
(	O
GSE	B-Organism_substance
)	O
,	O
a	O
widely	O
used	O
dietary	O
supplement	O
,	O
is	O
known	O
to	O
have	O
antitumor	B-Cancer
activity	O
.	O

In	O
this	O
study	O
,	O
we	O
have	O
explored	O
the	O
activity	O
of	O
GSE	B-Organism_substance
on	O
VEGF	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
and	O
angiogenesis	O
.	O

We	O
found	O
that	O
GSE	B-Organism_substance
could	O
directly	O
inhibit	O
the	O
kinase	O
activity	O
of	O
purified	O
VEGF	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
a	O
novel	O
activity	O
of	O
GSE	B-Organism_substance
that	O
has	O
not	O
been	O
characterized	O
.	O

GSE	B-Organism_substance
could	O
also	O
inhibit	O
the	O
VEGF	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
/	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
mediated	O
signaling	O
pathway	O
in	O
endothelial	B-Cell
cells	I-Cell
.	O

As	O
a	O
result	O
,	O
GSE	B-Organism_substance
could	O
inhibit	O
VEGF	B-Gene_or_gene_product
-	O
induced	O
endothelial	B-Cell
cell	I-Cell
proliferation	O
and	O
migration	O
as	O
well	O
as	O
sprout	B-Tissue
formation	O
from	O
aorta	B-Multi-tissue_structure
ring	I-Multi-tissue_structure
.	O

In	O
vivo	O
assay	O
further	O
showed	O
that	O
GSE	B-Organism_substance
could	O
inhibit	O
tumor	B-Cancer
growth	O
and	O
tumor	B-Cancer
angiogenesis	O
of	O
MDA	B-Cell
-	I-Cell
MB	I-Cell
-	I-Cell
231	I-Cell
breast	I-Cell
cancer	I-Cell
cells	I-Cell
in	O
mice	B-Organism
.	O

Consistent	O
with	O
the	O
in	O
vitro	O
data	O
,	O
GSE	B-Organism_substance
treatment	O
of	O
tumor	B-Cancer
-	O
bearing	O
mice	B-Organism
led	O
to	O
concomitant	O
reduction	O
of	O
blood	B-Multi-tissue_structure
vessel	I-Multi-tissue_structure
density	O
and	O
phosphorylation	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
kinase	I-Gene_or_gene_product
.	O

Depletion	O
of	O
polyphenol	B-Simple_chemical
with	O
polyvinylpyrrolidone	B-Simple_chemical
abolished	O
the	O
antiangiogenic	O
activity	O
of	O
GSE	B-Organism_substance
,	O
suggesting	O
a	O
water	O
-	O
soluble	O
fraction	O
of	O
polyphenol	B-Simple_chemical
in	O
GSE	B-Organism_substance
is	O
responsible	O
for	O
the	O
antiangiogenic	O
activity	O
.	O

Taken	O
together	O
,	O
this	O
study	O
indicates	O
that	O
GSE	B-Organism_substance
is	O
a	O
well	O
-	O
tolerated	O
and	O
inexpensive	O
natural	O
VEGF	B-Gene_or_gene_product
inhibitor	O
and	O
could	O
potentially	O
be	O
useful	O
in	O
cancer	B-Cancer
prevention	O
or	O
treatment	O
.	O

Role	O
of	O
the	O
interferon	B-Gene_or_gene_product
-	O
inducible	O
IFI16	B-Gene_or_gene_product
gene	O
in	O
the	O
induction	O
of	O
ICAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
by	O
TNF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
.	O

The	O
Interferon	B-Gene_or_gene_product
-	O
inducible	O
gene	O
IFI16	B-Gene_or_gene_product
,	O
a	O
member	O
of	O
the	O
HIN200	B-Gene_or_gene_product
family	O
,	O
is	O
activated	O
by	O
oxidative	O
stress	O
and	O
cell	B-Cell
density	O
,	O
in	O
addition	O
to	O
Interferons	B-Gene_or_gene_product
,	O
and	O
it	O
is	O
implicated	O
in	O
the	O
regulation	O
of	O
endothelial	B-Cell
cell	I-Cell
proliferation	O
and	O
vessel	B-Multi-tissue_structure
formation	O
in	O
vitro	O
.	O

We	O
have	O
previously	O
shown	O
that	O
IFI16	B-Gene_or_gene_product
is	O
required	O
for	O
proinflammatory	O
gene	O
stimulation	O
by	O
IFN	B-Gene_or_gene_product
-	I-Gene_or_gene_product
gamma	I-Gene_or_gene_product
through	O
the	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
complex	O
.	O

To	O
examine	O
whether	O
IFI16	B-Gene_or_gene_product
induction	O
might	O
be	O
extended	O
to	O
other	O
proinflammatory	O
cytokines	O
such	O
as	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
we	O
used	O
the	O
strategy	O
of	O
the	O
RNA	O
interference	O
to	O
knock	O
down	O
IFI16	B-Gene_or_gene_product
expression	O
,	O
and	O
analyze	O
the	O
capability	O
of	O
TNF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
to	O
stimulate	O
intercellular	B-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
ICAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
or	O
CD54	B-Gene_or_gene_product
)	O
expression	O
in	O
the	O
absence	O
of	O
functional	O
IFI16	B-Gene_or_gene_product
.	O

Our	O
studies	O
demonstrate	O
that	O
IFI16	B-Gene_or_gene_product
mediates	O
ICAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
stimulation	O
by	O
TNF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
through	O
the	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
pathway	O
,	O
thus	O
reinforcing	O
the	O
role	O
of	O
the	O
IFI16	B-Gene_or_gene_product
molecule	O
in	O
the	O
inflammation	O
process	O
.	O

Impact	O
of	O
tumor	B-Cell
cell	I-Cell
VEGF	B-Gene_or_gene_product
expression	O
on	O
the	O
in	O
vivo	O
efficacy	O
of	O
vandetanib	B-Simple_chemical
(	O
ZACTIMA	B-Simple_chemical
;	O
ZD6474	B-Simple_chemical
)	O
.	O

VEGF	B-Gene_or_gene_product
is	O
the	O
key	O
player	O
in	O
tumor	B-Cancer
angiogenesis	O
.	O

In	O
the	O
current	O
study	O
,	O
the	O
impact	O
of	O
VEGF	B-Gene_or_gene_product
expression	O
on	O
the	O
response	O
of	O
tumors	B-Cancer
to	O
the	O
VEGFR2	B-Gene_or_gene_product
associated	O
tyrosine	O
kinase	O
inhibitor	O
vandetanib	B-Simple_chemical
was	O
evaluated	O
.	O

MATERIALS	O
AND	O
METHODS	O
:	O
Human	B-Organism
colon	B-Cancer
carcinoma	I-Cancer
(	O
HT29	B-Cell
)	O
and	O
murine	B-Organism
squamous	B-Cancer
carcinoma	I-Cancer
(	O
SCCVII	B-Cell
)	O
clonal	O
cell	B-Cell
lines	I-Cell
expressing	O
varying	O
levels	O
of	O
VEGF	B-Gene_or_gene_product
were	O
established	O
and	O
their	O
response	O
to	O
vandetanib	B-Simple_chemical
was	O
assessed	O
in	O
tissue	B-Tissue
culture	O
and	O
as	O
solid	B-Cancer
tumors	I-Cancer
.	O

RESULTS	O
:	O
Vandetanib	B-Simple_chemical
treatment	O
had	O
no	O
effect	O
on	O
tumor	B-Cell
cell	I-Cell
clonogenic	O
cell	B-Cell
survival	O
in	O
vitro	O
but	O
doses	O
>	O
or	O
=	O
10	O
nM	O
significantly	O
reduced	O
endothelial	B-Cell
cell	I-Cell
migration	O
.	O

In	O
vivo	O
,	O
tumors	B-Cancer
derived	O
from	O
cell	B-Cell
clones	I-Cell
expressing	O
high	O
levels	O
of	O
VEGF	B-Gene_or_gene_product
displayed	O
significantly	O
enhanced	O
angiogenesis	O
and	O
more	O
aggressive	O
growth	O
.	O

An	O
intradermal	B-Multi-tissue_structure
angiogenesis	O
assay	O
was	O
used	O
to	O
demonstrate	O
that	O
a	O
4	O
-	O
day	O
treatment	O
with	O
vandetanib	B-Simple_chemical
(	O
50	O
mg	O
/	O
kg	O
/	O
day	O
)	O
was	O
able	O
to	O
significantly	O
inhibit	O
blood	B-Multi-tissue_structure
vessel	I-Multi-tissue_structure
growth	O
induced	O
by	O
both	O
parental	O
and	O
high	O
VEGF	B-Gene_or_gene_product
-	O
expressing	O
tumor	B-Cell
cell	I-Cell
clones	I-Cell
.	O

In	O
the	O
HT29	B-Cancer
tumor	I-Cancer
model	O
,	O
treatment	O
response	O
to	O
vandetanib	B-Simple_chemical
(	O
50	O
mg	O
/	O
kg	O
/	O
day	O
,	O
Monday	O
-	O
Friday	O
for	O
2	O
weeks	O
)	O
was	O
greatest	O
in	O
xenografts	B-Cancer
derived	O
from	O
the	O
highest	O
VEGF	B-Gene_or_gene_product
-	O
expressing	O
cell	B-Cell
clones	I-Cell
.	O

A	O
similar	O
trend	O
was	O
noted	O
in	O
the	O
SCCVII	B-Cancer
tumor	I-Cancer
model	O
.	O

The	O
present	O
findings	O
indicate	O
that	O
vandetanib	B-Simple_chemical
therapy	O
effectively	O
counteracted	O
the	O
aggressive	O
feature	O
of	O
tumor	B-Cancer
growth	O
resulting	O
from	O
VEGF	B-Gene_or_gene_product
over	O
-	O
expressing	O
tumor	B-Cell
cells	I-Cell
and	O
suggest	O
that	O
such	O
tumors	B-Cancer
may	O
be	O
particularly	O
well	O
suited	O
for	O
anti	O
-	O
VEGF	B-Gene_or_gene_product
interventions	O
.	O

c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Ski	I-Gene_or_gene_product
overexpression	O
promotes	O
tumor	B-Cancer
growth	O
and	O
angiogenesis	O
through	O
inhibition	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
in	O
diffuse	O
-	O
type	O
gastric	B-Cancer
carcinoma	I-Cancer
.	O

c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Ski	I-Gene_or_gene_product
,	O
originally	O
identified	O
as	O
a	O
proto	O
-	O
oncogene	O
product	O
,	O
is	O
an	O
important	O
negative	O
regulator	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
TGF	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
family	O
signaling	O
through	O
interaction	O
with	O
Smad2	B-Gene_or_gene_product
,	O
Smad3	B-Gene_or_gene_product
,	O
and	O
Smad4	B-Gene_or_gene_product
.	O

High	O
expression	O
of	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Ski	I-Gene_or_gene_product
has	O
been	O
found	O
in	O
some	O
cancers	B-Cancer
,	O
including	O
gastric	B-Cancer
cancer	I-Cancer
.	O

We	O
previously	O
showed	O
that	O
disruption	O
of	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
signaling	O
by	O
dominant	O
-	O
negative	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
a	O
diffuse	O
-	O
type	O
gastric	B-Cancer
carcinoma	I-Cancer
model	O
accelerated	O
tumor	B-Cancer
growth	O
through	O
induction	O
of	O
tumor	B-Cancer
angiogenesis	O
by	O
decreased	O
expression	O
of	O
the	O
anti	O
-	O
angiogenic	O
factor	O
thrombospondin	B-Gene_or_gene_product
(	I-Gene_or_gene_product
TSP	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

Here	O
,	O
we	O
examined	O
the	O
function	O
of	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Ski	I-Gene_or_gene_product
in	O
human	B-Organism
diffuse	O
-	O
type	O
gastric	B-Cancer
carcinoma	I-Cancer
OCUM	B-Cell
-	I-Cell
2MLN	I-Cell
cells	I-Cell
.	O

Overexpression	O
of	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Ski	I-Gene_or_gene_product
inhibited	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
signaling	O
in	O
OCUM	B-Cell
-	I-Cell
2MLN	I-Cell
cells	I-Cell
.	O

Interestingly	O
,	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Ski	I-Gene_or_gene_product
overexpression	O
resulted	O
in	O
extensive	O
acceleration	O
of	O
the	O
growth	O
of	O
subcutaneous	O
xenografts	B-Cancer
in	O
BALB	B-Organism
/	I-Organism
c	I-Organism
nu	I-Organism
/	I-Organism
nu	I-Organism
female	I-Organism
mice	I-Organism
(	O
6	O
weeks	O
of	O
age	O
)	O
.	O

Similar	O
to	O
tumors	B-Cancer
expressing	O
dominant	O
-	O
negative	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
,	O
histochemical	O
studies	O
revealed	O
less	O
fibrosis	O
and	O
increased	O
angiogenesis	O
in	O
xenografted	O
tumors	B-Cancer
expressing	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Ski	I-Gene_or_gene_product
compared	O
to	O
control	O
tumors	B-Cancer
.	O

Induction	O
of	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
mRNA	O
by	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
was	O
attenuated	O
by	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Ski	I-Gene_or_gene_product
in	O
vitro	O
,	O
and	O
expression	O
of	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
mRNA	O
was	O
decreased	O
in	O
tumors	B-Cancer
expressing	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Ski	I-Gene_or_gene_product
in	O
vivo	O
.	O

These	O
findings	O
suggest	O
that	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Ski	I-Gene_or_gene_product
overexpression	O
promotes	O
the	O
growth	O
of	O
diffuse	O
-	O
type	O
gastric	B-Cancer
carcinoma	I-Cancer
through	O
induction	O
of	O
angiogenesis	O
.	O

The	O
Down	B-Gene_or_gene_product
syndrome	I-Gene_or_gene_product
critical	I-Gene_or_gene_product
region	I-Gene_or_gene_product
gene	I-Gene_or_gene_product
1	I-Gene_or_gene_product
short	O
variant	O
promoters	O
direct	O
vascular	B-Multi-tissue_structure
bed	I-Multi-tissue_structure
-	O
specific	O
gene	O
expression	O
during	O
inflammation	O
in	O
mice	B-Organism
.	O

Down	B-Gene_or_gene_product
syndrome	I-Gene_or_gene_product
critical	I-Gene_or_gene_product
region	I-Gene_or_gene_product
gene	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
DSCR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
short	O
variant	O
(	O
DSCR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1s	I-Gene_or_gene_product
)	O
is	O
an	O
inhibitor	O
of	O
calcineurin	B-Gene_or_gene_product
/	O
NFAT	B-Gene_or_gene_product
signaling	O
encoded	O
by	O
exons	O
4	O
-	O
7	O
of	O
DSCR1	B-Gene_or_gene_product
.	O

We	O
previously	O
reported	O
that	O
VEGF	B-Gene_or_gene_product
induces	O
DSCR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1s	I-Gene_or_gene_product
expression	O
in	O
endothelial	B-Cell
cells	I-Cell
,	O
which	O
in	O
turn	O
negatively	O
feeds	O
back	O
to	O
attenuate	O
endothelial	B-Cell
cell	I-Cell
activation	O
.	O

Here	O
,	O
in	O
order	O
to	O
characterize	O
the	O
role	O
of	O
the	O
promoter	O
that	O
drives	O
DSCR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1s	I-Gene_or_gene_product
expression	O
in	O
mediating	O
inducible	O
expression	O
in	O
vivo	O
and	O
to	O
determine	O
the	O
functional	O
relevance	O
of	O
DSCR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1s	I-Gene_or_gene_product
in	O
inflammation	O
,	O
we	O
targeted	O
a	O
DNA	B-Cellular_component
construct	O
containing	O
1	O
.	O
7	O
kb	O
of	O
the	O
human	B-Organism
DSCR1s	B-Gene_or_gene_product
promoter	O
coupled	O
to	O
the	O
lacZ	B-Gene_or_gene_product
reporter	O
to	O
the	O
hypoxanthine	B-Gene_or_gene_product
guanine	I-Gene_or_gene_product
phosphoribosyl	I-Gene_or_gene_product
transferase	I-Gene_or_gene_product
(	O
Hprt	B-Gene_or_gene_product
)	O
locus	O
of	O
mice	B-Organism
.	O

We	O
determined	O
that	O
lacZ	B-Gene_or_gene_product
was	O
uniformly	O
expressed	O
in	O
the	O
endothelium	B-Tissue
of	O
transgenic	B-Developing_anatomical_structure
embryos	I-Developing_anatomical_structure
but	O
was	O
markedly	O
downregulated	O
postnatally	O
.	O

Systemic	O
administration	O
of	O
VEGF	B-Gene_or_gene_product
or	O
LPS	B-Simple_chemical
in	O
adult	O
mice	B-Organism
resulted	O
in	O
cyclosporine	B-Simple_chemical
A	I-Simple_chemical
-	O
sensitive	O
reactivation	O
of	O
the	O
DSCR1s	B-Gene_or_gene_product
promoter	O
and	O
endogenous	O
gene	O
expression	O
in	O
a	O
subset	O
of	O
organs	B-Organ
,	O
including	O
the	O
heart	B-Organ
and	O
brain	B-Organ
.	O

The	O
DSCR1s	B-Gene_or_gene_product
promoter	O
was	O
similarly	O
induced	O
in	O
the	O
endothelium	B-Tissue
of	O
tumor	B-Cancer
xenografts	I-Cancer
.	O

In	O
a	O
mouse	B-Organism
model	O
of	O
endotoxemia	O
,	O
DSCR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1s	I-Gene_or_gene_product
-	O
deficient	O
mice	B-Organism
demonstrated	O
increased	O
sepsis	O
mortality	O
,	O
whereas	O
adenovirus	B-Organism
-	O
mediated	O
DSCR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1s	I-Gene_or_gene_product
overexpression	O
protected	O
against	O
LPS	B-Simple_chemical
-	O
induced	O
lethality	O
.	O

Collectively	O
,	O
these	O
data	O
suggest	O
that	O
the	O
DSCR1s	B-Gene_or_gene_product
promoter	O
directs	O
vascular	B-Multi-tissue_structure
bed	I-Multi-tissue_structure
-	O
specific	O
expression	O
in	O
activated	O
endothelium	B-Tissue
and	O
that	O
DSCR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1s	I-Gene_or_gene_product
serves	O
to	O
dampen	O
the	O
host	O
response	O
to	O
infection	O
.	O

Nicked	B-Gene_or_gene_product
{	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
}	I-Gene_or_gene_product
2	I-Gene_or_gene_product
-	I-Gene_or_gene_product
glycoprotein	I-Gene_or_gene_product
I	I-Gene_or_gene_product
binds	O
angiostatin	B-Gene_or_gene_product
4	I-Gene_or_gene_product
.	I-Gene_or_gene_product
5	I-Gene_or_gene_product
(	O
plasminogen	O
kringle	O
1	O
-	O
5	O
)	O
and	O
attenuates	O
its	O
antiangiogenic	O
property	O
.	O

Angiostatin	B-Gene_or_gene_product
was	O
first	O
discovered	O
as	O
a	O
plasminogen	B-Gene_or_gene_product
fragment	I-Gene_or_gene_product
with	O
antitumor	B-Cancer
/	O
antiangiogenic	O
property	O
.	O

One	O
of	O
the	O
angiostatin	B-Gene_or_gene_product
isoforms	O
,	O
that	O
is	O
,	O
angiostatin	B-Gene_or_gene_product
4	I-Gene_or_gene_product
.	I-Gene_or_gene_product
5	I-Gene_or_gene_product
(	O
AS4	B-Gene_or_gene_product
.	I-Gene_or_gene_product
5	I-Gene_or_gene_product
)	O
,	O
consisting	O
of	O
plasminogen	O
kringle	O
1	O
to	O
4	O
and	O
a	O
most	O
part	O
of	O
kringle	O
5	O
,	O
is	O
produced	O
by	O
autoproteolysis	O
and	O
present	O
in	O
human	B-Organism
plasma	B-Organism_substance
.	O

beta2	B-Gene_or_gene_product
-	I-Gene_or_gene_product
glycoprotein	I-Gene_or_gene_product
I	I-Gene_or_gene_product
(	O
beta2GPI	B-Gene_or_gene_product
)	O
is	O
proteolytically	O
cleaved	O
by	O
plasmin	B-Gene_or_gene_product
in	O
its	O
domain	O
V	O
(	O
nicked	B-Gene_or_gene_product
beta2GPI	I-Gene_or_gene_product
)	O
,	O
resulting	O
in	O
binding	O
to	O
plasminogen	B-Gene_or_gene_product
.	O

Antiangiogenic	O
properties	O
have	O
been	O
recently	O
reported	O
in	O
nicked	B-Gene_or_gene_product
beta2GPI	I-Gene_or_gene_product
as	O
well	O
as	O
in	O
intact	O
beta2GPI	B-Gene_or_gene_product
at	O
higher	O
concentrations	O
.	O

In	O
the	O
present	O
study	O
,	O
we	O
found	O
significant	O
binding	O
of	O
nicked	B-Gene_or_gene_product
beta2GPI	I-Gene_or_gene_product
to	O
AS4	B-Gene_or_gene_product
.	I-Gene_or_gene_product
5	I-Gene_or_gene_product
(	O
K	O
(	O
D	O
)	O
=	O
3	O
.	O
27	O
x	O
10	O
(	O
6	O
)	O
M	O
(	O
-	O
1	O
)	O
)	O
.	O

Via	O
this	O
binding	O
,	O
nicked	B-Gene_or_gene_product
beta2GPI	I-Gene_or_gene_product
attenuates	O
the	O
antiangiogenic	O
functions	O
of	O
AS4	B-Gene_or_gene_product
.	I-Gene_or_gene_product
5	I-Gene_or_gene_product
in	O
the	O
proliferation	O
of	O
arterial	B-Cell
/	O
venous	B-Cell
endothelial	I-Cell
cells	I-Cell
,	O
in	O
the	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
invasion	O
and	O
the	O
tube	B-Tissue
formation	O
of	O
venous	B-Cell
endothelial	I-Cell
cells	I-Cell
,	O
and	O
in	O
vivo	O
angiogenesis	O
.	O

In	O
contrast	O
,	O
intact	O
beta2GPI	B-Gene_or_gene_product
does	O
not	O
bind	O
to	O
AS4	B-Gene_or_gene_product
.	I-Gene_or_gene_product
5	I-Gene_or_gene_product
or	O
inhibit	O
its	O
antiangiogenic	O
activity	O
.	O

Thus	O
,	O
nicked	B-Gene_or_gene_product
beta2GPI	I-Gene_or_gene_product
exerts	O
dual	O
effects	O
on	O
angiogenesis	O
,	O
that	O
is	O
,	O
nicked	B-Gene_or_gene_product
beta2GPI	I-Gene_or_gene_product
promotes	O
angiogenesis	O
in	O
the	O
presence	O
of	O
AS4	B-Gene_or_gene_product
.	I-Gene_or_gene_product
5	I-Gene_or_gene_product
,	O
whereas	O
nicked	B-Gene_or_gene_product
beta2GPI	I-Gene_or_gene_product
inhibits	O
angiogenesis	O
at	O
concentrations	O
high	O
enough	O
to	O
neutralize	O
AS4	B-Gene_or_gene_product
.	I-Gene_or_gene_product
5	I-Gene_or_gene_product
.	O

Our	O
data	O
suggest	O
that	O
plasmin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
nicked	I-Gene_or_gene_product
beta2GPI	I-Gene_or_gene_product
promotes	O
angiogenesis	O
by	O
interacting	O
with	O
plasmin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
generated	I-Gene_or_gene_product
AS4	I-Gene_or_gene_product
.	I-Gene_or_gene_product
5	I-Gene_or_gene_product
in	O
sites	O
of	O
increased	O
fibrinolysis	O
such	O
as	O
thrombus	B-Pathological_formation
.	O

[	O
An	O
experimental	O
study	O
on	O
angiogenesis	O
of	O
non	O
-	O
vascularized	O
autogenous	B-Multi-tissue_structure
bone	I-Multi-tissue_structure
graft	I-Multi-tissue_structure
with	O
vascular	B-Multi-tissue_structure
bundle	I-Multi-tissue_structure
implantation	O
]	O

OBJECTIVE	O
:	O
To	O
investigate	O
the	O
effect	O
of	O
vascular	B-Multi-tissue_structure
bundle	I-Multi-tissue_structure
implantation	O
in	O
autogenous	B-Multi-tissue_structure
bone	I-Multi-tissue_structure
graft	I-Multi-tissue_structure
on	O
angiogenesis	O
.	O

METHODS	O
:	O
Thirty	O
-	O
six	O
New	B-Organism
Zealand	I-Organism
white	I-Organism
rabbits	I-Organism
were	O
evaluated	O
in	O
this	O
study	O
.	O

A	O
portion	O
of	O
bilateral	B-Organ
radial	I-Organ
bones	I-Organ
of	O
a	O
rabbit	B-Organism
were	O
removed	O
as	O
free	O
bone	B-Multi-tissue_structure
grafts	I-Multi-tissue_structure
,	O
whose	O
periostea	B-Tissue
were	O
peeled	O
off	O
.	O

In	O
test	O
group	O
,	O
the	O
external	B-Multi-tissue_structure
maxillary	I-Multi-tissue_structure
artery	I-Multi-tissue_structure
bundle	I-Multi-tissue_structure
was	O
passed	O
through	O
the	O
marrow	B-Immaterial_anatomical_entity
cavity	I-Immaterial_anatomical_entity
of	O
the	O
bone	B-Organ
.	O

In	O
control	O
group	O
,	O
there	O
was	O
no	O
vascular	B-Multi-tissue_structure
bundle	I-Multi-tissue_structure
implantation	O
.	O

Each	O
bone	B-Organ
was	O
placed	O
in	O
masseter	B-Organ
muscle	I-Organ
separately	O
.	O

The	O
rabbits	B-Organism
were	O
sacrificed	O
and	O
the	O
specimens	B-Multi-tissue_structure
were	O
procured	O
at	O
3	O
days	O
,	O
1	O
,	O
2	O
,	O
3	O
,	O
4	O
and	O
6	O
weeks	O
after	O
surgery	O
for	O
histological	O
observation	O
,	O
Chinese	O
ink	O
perfusion	O
and	O
CD34	B-Gene_or_gene_product
immunohistochemistry	O
.	O

Microvessel	B-Tissue
density	O
(	O
MVD	O
)	O
was	O
assessed	O
in	O
order	O
to	O
evaluate	O
angiogenesis	O
of	O
autogenous	B-Multi-tissue_structure
bone	I-Multi-tissue_structure
grafts	I-Multi-tissue_structure
.	O

RESULTS	O
:	O
The	O
bone	B-Multi-tissue_structure
grafts	I-Multi-tissue_structure
were	O
found	O
revascularization	O
in	O
3	O
days	O
after	O
surgery	O
in	O
the	O
test	O
group	O
,	O
whereas	O
at	O
2	O
weeks	O
in	O
the	O
control	O
group	O
.	O

In	O
3	O
days	O
,	O
1	O
week	O
,	O
2	O
weeks	O
,	O
3	O
weeks	O
and	O
4	O
weeks	O
after	O
surgery	O
,	O
the	O
MVD	O
of	O
test	O
group	O
was	O
significantly	O
higher	O
than	O
that	O
of	O
control	O
group	O
.	O

In	O
4	O
weeks	O
after	O
surgery	O
,	O
angiogenesis	O
of	O
test	O
group	O
reached	O
to	O
peak	O
.	O

CONCLUSION	O
:	O
Vascular	B-Multi-tissue_structure
bundle	I-Multi-tissue_structure
implantation	O
improved	O
angiogenesis	O
in	O
non	O
-	O
vascularized	O
autogenous	B-Multi-tissue_structure
bone	I-Multi-tissue_structure
graft	I-Multi-tissue_structure
in	O
this	O
study	O
.	O

Targeting	O
glucose	B-Simple_chemical
consumption	O
and	O
autophagy	O
in	O
myeloma	B-Cancer
with	O
the	O
novel	O
nucleoside	B-Simple_chemical
analogue	I-Simple_chemical
8	I-Simple_chemical
-	I-Simple_chemical
aminoadenosine	I-Simple_chemical
.	O

Multiple	O
myeloma	B-Cancer
,	O
an	O
incurable	O
plasma	B-Cancer
cell	I-Cancer
malignancy	I-Cancer
,	O
is	O
characterized	O
by	O
altered	O
cellular	B-Cell
metabolism	O
and	O
resistance	O
to	O
apoptosis	O
.	O

Recent	O
connections	O
between	O
glucose	B-Simple_chemical
metabolism	O
and	O
resistance	O
to	O
apoptosis	O
provide	O
a	O
compelling	O
rationale	O
for	O
targeting	O
metabolic	O
changes	O
in	O
cancer	B-Cancer
.	O

In	O
this	O
study	O
,	O
we	O
have	O
examined	O
the	O
ability	O
of	O
the	O
purine	B-Simple_chemical
analogue	I-Simple_chemical
8	I-Simple_chemical
-	I-Simple_chemical
aminoadenosine	I-Simple_chemical
to	O
acutely	O
reduce	O
glucose	B-Simple_chemical
consumption	O
by	O
regulating	O
localization	O
and	O
expression	O
of	O
key	O
glucose	B-Gene_or_gene_product
transporters	I-Gene_or_gene_product
.	O

Myeloma	B-Cell
cells	I-Cell
counteracted	O
the	O
metabolic	O
stress	O
by	O
activating	O
autophagy	O
.	O

Co	O
-	O
treatment	O
with	O
inhibitors	O
of	O
autophagy	O
results	O
in	O
marked	O
enhancement	O
of	O
cell	B-Cell
death	O
.	O

Glucose	B-Simple_chemical
consumption	O
by	O
drug	O
-	O
resistant	O
myeloma	B-Cell
cells	I-Cell
was	O
unaffected	O
by	O
8	B-Simple_chemical
-	I-Simple_chemical
aminoadenosine	I-Simple_chemical
,	O
and	O
accordingly	O
,	O
no	O
activation	O
of	O
autophagy	O
was	O
observed	O
.	O

However	O
,	O
these	O
cells	B-Cell
can	O
be	O
sensitized	O
to	O
8	B-Simple_chemical
-	I-Simple_chemical
aminoadenosine	I-Simple_chemical
under	O
glucose	B-Simple_chemical
-	O
limiting	O
conditions	O
.	O

The	O
prosurvival	O
autophagic	O
response	O
of	O
myeloma	B-Cancer
to	O
nutrient	O
deprivation	O
or	O
to	O
nucleoside	B-Simple_chemical
analogue	I-Simple_chemical
treatment	O
has	O
not	O
been	O
described	O
previously	O
.	O

This	O
study	O
establishes	O
the	O
potential	O
of	O
metabolic	O
targeting	O
as	O
a	O
broader	O
means	O
to	O
kill	O
and	O
sensitize	O
myeloma	B-Cancer
and	O
identifies	O
a	O
compound	O
that	O
can	O
achieve	O
this	O
goal	O
.	O

Dopamine	B-Simple_chemical
regulates	O
phosphorylation	O
of	O
VEGF	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
2	I-Gene_or_gene_product
by	O
engaging	O
Src	B-Gene_or_gene_product
-	I-Gene_or_gene_product
homology	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
-	I-Gene_or_gene_product
domain	I-Gene_or_gene_product
-	I-Gene_or_gene_product
containing	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
tyrosine	I-Gene_or_gene_product
phosphatase	I-Gene_or_gene_product
2	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
VEGF	B-Gene_or_gene_product
)	O
-	O
induced	O
receptor	O
phosphorylation	O
is	O
the	O
crucial	O
step	O
for	O
initiating	O
downstream	O
signaling	O
pathways	O
that	O
lead	O
to	O
angiogenesis	O
or	O
related	O
pathophysiological	O
outcomes	O
.	O

Our	O
previous	O
studies	O
have	O
shown	O
that	O
the	O
neurotransmitter	O
dopamine	B-Simple_chemical
could	O
inhibit	O
VEGF	B-Gene_or_gene_product
-	O
induced	O
phosphorylation	O
of	O
VEGF	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
2	I-Gene_or_gene_product
(	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
)	O
,	O
endothelial	B-Cell
cell	I-Cell
proliferation	O
,	O
migration	O
,	O
microvascular	B-Tissue
permeability	O
,	O
and	O
thus	O
,	O
angiogenesis	O
.	O

In	O
this	O
study	O
,	O
we	O
address	O
the	O
mechanism	O
by	O
which	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
phosphorylation	O
is	O
regulated	O
by	O
dopamine	B-Simple_chemical
.	O

Here	O
,	O
we	O
demonstrate	O
that	O
D2	B-Gene_or_gene_product
dopamine	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
(	O
D2DR	B-Gene_or_gene_product
)	O
colocalizes	O
with	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
at	O
the	O
cell	B-Cellular_component
surface	I-Cellular_component
.	O

Dopamine	B-Simple_chemical
pretreatment	O
increases	O
the	O
translocation	O
and	O
colocalization	O
of	O
Src	B-Gene_or_gene_product
-	I-Gene_or_gene_product
homology	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
-	I-Gene_or_gene_product
domain	I-Gene_or_gene_product
-	I-Gene_or_gene_product
containing	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
tyrosine	I-Gene_or_gene_product
phosphatase	I-Gene_or_gene_product
(	O
SHP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
)	O
with	O
D2DR	B-Gene_or_gene_product
at	O
the	O
cell	B-Cellular_component
surface	I-Cellular_component
.	O

Dopamine	B-Simple_chemical
administration	O
leads	O
to	O
increased	O
VEGF	B-Gene_or_gene_product
-	O
induced	O
phosphorylation	O
of	O
SHP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
this	O
increased	O
phosphorylation	O
parallels	O
the	O
increased	O
phosphatase	O
activity	O
of	O
SHP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
.	O

Active	O
SHP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
then	O
dephosphorylates	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
at	O
Y951	O
,	O
Y996	O
and	O
Y1059	O
,	O
but	O
not	O
Y1175	O
.	O

We	O
also	O
observe	O
that	O
SHP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
knockdown	O
impairs	O
the	O
dopamine	B-Simple_chemical
-	O
regulated	O
inhibition	O
of	O
VEGF	B-Gene_or_gene_product
-	O
induced	O
phosphorylation	O
of	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
,	O
subsequently	O
,	O
Src	B-Gene_or_gene_product
phosphorylation	O
and	O
migration	O
.	O

Our	O
data	O
establish	O
a	O
novel	O
role	O
for	O
SHP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
phosphatase	I-Gene_or_gene_product
in	O
the	O
dopamine	B-Simple_chemical
-	O
mediated	O
regulation	O
of	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
phosphorylation	O
.	O

Glioma	B-Cell
tumor	I-Cell
stem	I-Cell
-	I-Cell
like	I-Cell
cells	I-Cell
promote	O
tumor	B-Cancer
angiogenesis	O
and	O
vasculogenesis	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
and	O
stromal	B-Gene_or_gene_product
-	I-Gene_or_gene_product
derived	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

Cancer	B-Cell
stem	I-Cell
cells	I-Cell
(	O
CSC	B-Cell
)	O
are	O
predicted	O
to	O
be	O
critical	O
drivers	O
of	O
tumor	B-Cancer
progression	O
due	O
to	O
their	O
self	O
-	O
renewal	O
capacity	O
and	O
limitless	O
proliferative	O
potential	O
.	O

An	O
emerging	O
area	O
of	O
research	O
suggests	O
that	O
CSC	B-Cell
may	O
also	O
support	O
tumor	B-Cancer
progression	O
by	O
promoting	O
tumor	B-Cancer
angiogenesis	O
.	O

To	O
investigate	O
how	O
CSC	B-Cell
contribute	O
to	O
tumor	B-Multi-tissue_structure
vascular	I-Multi-tissue_structure
development	O
,	O
we	O
used	O
an	O
approach	O
comparing	O
tumor	B-Cancer
xenografts	I-Cancer
of	O
the	O
C6	B-Cell
glioma	I-Cell
cell	I-Cell
line	I-Cell
containing	O
either	O
a	O
low	O
or	O
a	O
high	O
fraction	O
of	O
CSC	B-Cell
.	O

Compared	O
with	O
CSC	B-Cancer
-	I-Cancer
low	I-Cancer
tumors	I-Cancer
,	O
CSC	B-Cancer
-	I-Cancer
high	I-Cancer
tumors	I-Cancer
exhibited	O
increased	O
microvessel	B-Tissue
density	O
and	O
blood	B-Organism_substance
perfusion	O
and	O
induced	O
increased	O
mobilization	O
and	O
tumor	B-Cancer
recruitment	O
of	O
bone	B-Multi-tissue_structure
marrow	I-Multi-tissue_structure
-	O
derived	O
endothelial	B-Cell
progenitor	I-Cell
cells	I-Cell
(	O
EPC	B-Cell
)	O
.	O

CSC	B-Cell
-	I-Cell
high	I-Cell
C6	I-Cell
cell	I-Cell
cultures	I-Cell
also	O
induced	O
higher	O
levels	O
of	O
endothelial	B-Cell
cell	I-Cell
proliferation	O
and	O
tubule	B-Tissue
organization	O
in	O
vitro	O
compared	O
with	O
CSC	B-Cell
-	I-Cell
low	I-Cell
cultures	I-Cell
.	O

CSC	B-Cell
-	I-Cell
high	I-Cell
cultures	I-Cell
and	O
tumors	B-Cancer
expressed	O
increased	O
levels	O
of	O
the	O
proangiogenic	O
factors	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
stromal	B-Gene_or_gene_product
-	I-Gene_or_gene_product
derived	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
and	O
when	O
signaling	O
by	O
either	O
factor	O
was	O
blocked	O
,	O
all	O
aspects	O
of	O
angiogenesis	O
observed	O
in	O
CSC	B-Cell
-	I-Cell
high	I-Cell
cultures	I-Cell
and	O
tumors	B-Cancer
,	O
including	O
microvessel	B-Tissue
density	O
,	O
perfusion	O
,	O
EPC	B-Cell
mobilization	O
/	O
recruitment	O
,	O
and	O
stimulation	O
of	O
endothelial	B-Cell
cell	I-Cell
activity	O
,	O
were	O
reduced	O
to	O
levels	O
comparable	O
with	O
those	O
observed	O
in	O
CSC	B-Cell
-	I-Cell
low	I-Cell
cultures	I-Cell
/	O
tumors	B-Cancer
.	O

These	O
results	O
suggest	O
that	O
CSC	B-Cell
contribute	O
to	O
tumor	B-Cancer
angiogenesis	O
by	O
promoting	O
both	O
local	O
endothelial	B-Cell
cell	I-Cell
activity	O
and	O
systemic	O
angiogenic	O
processes	O
involving	O
bone	B-Multi-tissue_structure
marrow	I-Multi-tissue_structure
-	O
derived	O
EPC	B-Cell
in	O
a	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
dependent	O
and	O
stromal	B-Gene_or_gene_product
-	I-Gene_or_gene_product
derived	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
dependent	O
manner	O
.	O

Inducible	B-Gene_or_gene_product
nitric	I-Gene_or_gene_product
oxide	I-Gene_or_gene_product
synthase	I-Gene_or_gene_product
modulates	O
angiogenesis	O
in	O
ischemic	O
hindlimb	B-Organism_subdivision
of	O
rat	B-Organism
.	O

BACKGROUND	O
:	O
Angiogenesis	O
plays	O
an	O
important	O
role	O
in	O
maintaining	O
adequate	O
oxygen	B-Simple_chemical
delivery	O
,	O
and	O
nitric	B-Simple_chemical
oxide	I-Simple_chemical
(	O
NO	B-Simple_chemical
)	O
is	O
a	O
potential	O
regulator	O
of	O
angiogenesis	O
.	O

NO	B-Simple_chemical
is	O
synthesized	O
through	O
three	O
isoforms	O
of	O
NO	B-Gene_or_gene_product
synthase	I-Gene_or_gene_product
(	O
NOS	B-Gene_or_gene_product
)	O
.	O

It	O
is	O
hypothesized	O
that	O
the	O
NO	B-Simple_chemical
derived	O
from	O
inducible	B-Gene_or_gene_product
NOS	I-Gene_or_gene_product
(	O
iNOS	B-Gene_or_gene_product
)	O
may	O
promote	O
survival	O
of	O
ischemic	B-Tissue
tissue	I-Tissue
through	O
angiogenesis	O
.	O

To	O
test	O
this	O
hypothesis	O
,	O
we	O
investigated	O
the	O
effect	O
of	O
iNOS	B-Gene_or_gene_product
deficiency	O
(	O
by	O
L	B-Simple_chemical
-	I-Simple_chemical
NIL	I-Simple_chemical
)	O
on	O
angiogenesis	O
in	O
a	O
hindlimb	B-Organism_subdivision
ischemia	O
model	O
.	O

METHODS	O
:	O
Thirty	O
-	O
two	O
male	O
wistar	B-Organism
rats	I-Organism
randomly	O
divided	O
into	O
four	O
groups	O
.	O

In	O
groups	O
1	O
&	O
2	O
,	O
hindlimb	B-Organism_subdivision
ischemia	O
was	O
induced	O
by	O
ligation	O
of	O
femoral	B-Multi-tissue_structure
artery	I-Multi-tissue_structure
and	O
they	O
received	O
L	B-Simple_chemical
-	I-Simple_chemical
NIL	I-Simple_chemical
and	O
saline	B-Simple_chemical
respectively	O
.	O

The	O
animals	O
in	O
groups	O
3	O
and	O
4	O
also	O
received	O
L	B-Simple_chemical
-	I-Simple_chemical
NIL	I-Simple_chemical
and	O
saline	B-Simple_chemical
respectively	O
without	O
surgical	O
procedure	O
.	O

After	O
21	O
days	O
,	O
the	O
serum	B-Organism_substance
concentration	O
of	O
nitrite	B-Simple_chemical
,	O
capillary	B-Tissue
density	O
and	O
expression	O
of	O
HIF1alpha	B-Gene_or_gene_product
were	O
determined	O
.	O

RESULTS	O
:	O
Serum	B-Organism_substance
nitrite	B-Simple_chemical
levels	O
were	O
significantly	O
lower	O
in	O
L	B-Simple_chemical
-	I-Simple_chemical
NIL	I-Simple_chemical
groups	O
(	O
p	O
<	O
0	O
.	O
05	O
)	O
.	O

The	O
capillary	B-Tissue
density	O
in	O
group	O
1	O
(	O
ischemia	O
+	O
L	B-Simple_chemical
-	I-Simple_chemical
NIL	I-Simple_chemical
)	O
was	O
significantly	O
different	O
from	O
group	O
2	O
(	O
ischemia	O
+	O
saline	B-Simple_chemical
)	O
;	O
group	O
1	O
:	O
360	O
.	O
33	O
+	O
/	O
-	O
77	O
.	O
02	O
,	O
group	O
2	O
:	O
549	O
+	O
/	O
-	O
81	O
.	O
85	O
/	O
mm2	O
,	O
p	O
<	O
0	O
.	O
05	O
)	O
.	O
In	O
addition	O
,	O
expression	O
of	O
HIF1alpha	B-Gene_or_gene_product
was	O
significantly	O
increased	O
in	O
ischemic	O
groups	O
(	O
p	O
<	O
0	O
.	O
05	O
)	O
.	O

CONCLUSION	O
:	O
Selective	O
inhibition	O
of	O
iNOS	B-Gene_or_gene_product
by	O
L	B-Simple_chemical
-	I-Simple_chemical
NIL	I-Simple_chemical
inhibits	O
angiogenesis	O
in	O
a	O
hindlimb	B-Organism_subdivision
ischemic	O
rat	B-Organism
model	O
.	O

In	O
addition	O
,	O
ischemia	O
induces	O
expression	O
of	O
HIF1alpha	B-Gene_or_gene_product
in	O
hypoxic	B-Tissue
tissue	I-Tissue
.	O

Phenolic	O
fraction	O
of	O
tobacco	B-Simple_chemical
smoke	I-Simple_chemical
condensate	I-Simple_chemical
potentiates	O
benzo	B-Simple_chemical
[	I-Simple_chemical
a	I-Simple_chemical
]	I-Simple_chemical
pyerene	I-Simple_chemical
diol	I-Simple_chemical
epoxide	I-Simple_chemical
-	O
induced	O
cell	B-Cell
transformation	O
:	O
role	O
of	O
protein	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
C	I-Gene_or_gene_product
.	O

In	O
this	O
study	O
we	O
separated	O
weakly	O
acidic	O
phenolic	O
components	O
from	O
other	O
neutral	O
,	O
acidic	O
and	O
basic	O
components	O
of	O
tobacco	B-Simple_chemical
smoke	I-Simple_chemical
condensate	I-Simple_chemical
(	O
TSC	B-Simple_chemical
)	O
and	O
observed	O
that	O
phenolic	B-Simple_chemical
fraction	I-Simple_chemical
of	O
TSC	B-Simple_chemical
significantly	O
increased	O
the	O
number	O
of	O
colonies	O
of	O
promotion	O
-	O
sensitive	O
JB6	B-Cell
Cl41	I-Cell
cells	I-Cell
that	O
showed	O
anchorage	O
-	O
independent	O
growth	O
on	O
soft	O
agar	O
in	O
response	O
to	O
BPDE	B-Simple_chemical
(	O
an	O
ultimate	O
carcinogen	O
produced	O
by	O
metabolic	O
activation	O
of	O
the	O
PAH	B-Simple_chemical
benzo	I-Simple_chemical
[	I-Simple_chemical
a	I-Simple_chemical
]	I-Simple_chemical
pyrene	I-Simple_chemical
)	O
.	O

Anchorage	O
-	O
independent	O
cell	B-Cell
growth	O
is	O
indicative	O
of	O
cell	B-Cell
transformation	O
resulting	O
in	O
acquisition	O
of	O
tumorigenic	O
potential	O
.	O

In	O
order	O
to	O
understand	O
the	O
underlying	O
mechanism	O
by	O
which	O
TSC	B-Simple_chemical
phenolic	I-Simple_chemical
fraction	I-Simple_chemical
potentiates	O
BPDE	B-Simple_chemical
-	O
induced	O
tumorigenicity	O
,	O
we	O
examined	O
its	O
effect	O
on	O
the	O
activation	O
of	O
two	O
transcription	O
factors	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
kappaB	I-Gene_or_gene_product
which	O
are	O
known	O
to	O
be	O
influenced	O
by	O
established	O
tumor	B-Cancer
promoter	O
TPA	B-Simple_chemical
.	O

BPDE	B-Simple_chemical
treatment	O
caused	O
induction	O
of	O
both	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
kappaB	I-Gene_or_gene_product
activity	O
as	O
determined	O
by	O
luciferase	B-Gene_or_gene_product
reporter	O
assay	O
and	O
only	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
induction	O
in	O
response	O
to	O
BPDE	B-Simple_chemical
was	O
significantly	O
attenuated	O
by	O
TSC	B-Simple_chemical
phenolic	I-Simple_chemical
fraction	I-Simple_chemical
whereas	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
induction	O
remains	O
unaltered	O
.	O

Attenuation	O
of	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
activation	O
by	O
TSC	B-Simple_chemical
phenolic	I-Simple_chemical
fraction	I-Simple_chemical
was	O
associated	O
with	O
significant	O
decrease	O
of	O
intracellular	B-Immaterial_anatomical_entity
PKC	B-Gene_or_gene_product
substrate	O
phosphorylation	O
in	O
BPDE	B-Simple_chemical
treated	O
cells	B-Cell
.	O

Non	O
-	O
specific	O
PKC	B-Gene_or_gene_product
inhibitors	O
staurosporine	B-Simple_chemical
and	O
bisindolylmaleimide	B-Simple_chemical
II	I-Simple_chemical
as	O
well	O
as	O
inhibitors	O
specific	O
to	O
conventional	O
PKCs	B-Gene_or_gene_product
(	O
Go6976	B-Simple_chemical
)	O
and	O
PKC	B-Gene_or_gene_product
-	I-Gene_or_gene_product
delta	I-Gene_or_gene_product
(	O
rottlerin	B-Simple_chemical
)	O
attenuated	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
activation	O
in	O
BPDE	B-Simple_chemical
treated	O
cells	B-Cell
to	O
a	O
varying	O
degree	O
indicating	O
a	O
possible	O
link	O
between	O
PKC	B-Gene_or_gene_product
down	O
-	O
regulation	O
and	O
the	O
attenuation	O
of	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
activity	O
by	O
TSC	B-Simple_chemical
phenolic	I-Simple_chemical
fraction	I-Simple_chemical
.	O

Treatment	O
of	O
cells	B-Cell
with	O
PKC	B-Gene_or_gene_product
inhibitors	O
also	O
potentiated	O
anchorage	O
-	O
independent	O
growth	O
of	O
BPDE	B-Simple_chemical
treated	O
cells	B-Cell
on	O
soft	O
agar	O
.	O

Our	O
data	O
suggest	O
a	O
possible	O
role	O
of	O
PKC	B-Gene_or_gene_product
down	O
-	O
regulation	O
in	O
potentiation	O
of	O
BPDE	B-Simple_chemical
-	O
induced	O
tumorogenicity	O
by	O
TSC	B-Simple_chemical
phenolic	I-Simple_chemical
fraction	I-Simple_chemical
.	O

Killing	O
tumor	B-Cell
cells	I-Cell
through	O
their	O
surface	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
or	O
major	B-Gene_or_gene_product
histocompatibility	I-Gene_or_gene_product
complex	I-Gene_or_gene_product
class	I-Gene_or_gene_product
I	I-Gene_or_gene_product
molecules	O
.	O

Targeted	O
antibody	O
-	O
based	O
therapy	O
has	O
been	O
used	O
successfully	O
to	O
treat	O
cancers	B-Cancer
.	O

Recent	O
studies	O
have	O
demonstrated	O
that	O
tumor	B-Cell
cells	I-Cell
treated	O
with	O
antibodies	O
specific	O
for	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
or	O
major	B-Gene_or_gene_product
histocompatibility	I-Gene_or_gene_product
complex	I-Gene_or_gene_product
(	I-Gene_or_gene_product
MHC	I-Gene_or_gene_product
)	I-Gene_or_gene_product
class	I-Gene_or_gene_product
I	I-Gene_or_gene_product
molecules	O
undergo	O
apoptosis	O
in	O
vitro	O
and	O
in	O
vivo	O
(	O
mouse	B-Organism
models	O
)	O
.	O

Antibodies	O
against	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
or	O
MHC	B-Gene_or_gene_product
class	I-Gene_or_gene_product
I	I-Gene_or_gene_product
induce	O
tumor	B-Cell
cell	I-Cell
apoptosis	O
by	O
1	O
)	O
recruiting	O
MHC	B-Gene_or_gene_product
class	I-Gene_or_gene_product
I	I-Gene_or_gene_product
molecules	O
to	O
lipid	B-Cellular_component
rafts	I-Cellular_component
and	O
activating	O
LYN	B-Gene_or_gene_product
kinase	O
and	O
the	O
signal	O
-	O
transducing	O
enzyme	O
phospholipase	B-Gene_or_gene_product
C	I-Gene_or_gene_product
-	I-Gene_or_gene_product
gamma2	I-Gene_or_gene_product
-	O
dependent	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
signaling	O
pathway	O
and	O
2	O
)	O
expelling	O
interleukin	B-Gene_or_gene_product
6	I-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
1	I-Gene_or_gene_product
receptors	I-Gene_or_gene_product
out	O
of	O
lipid	B-Cellular_component
rafts	I-Cellular_component
and	O
inhibiting	O
the	O
growth	O
and	O
survival	O
factor	O
-	O
induced	O
activation	O
of	O
the	O
phosphatidylinositol	B-Gene_or_gene_product
3	I-Gene_or_gene_product
-	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
/	O
Akt	B-Gene_or_gene_product
and	O
extracellular	B-Gene_or_gene_product
signal	I-Gene_or_gene_product
-	I-Gene_or_gene_product
related	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
pathways	O
.	O

Consequently	O
,	O
mitochondrial	B-Cellular_component
integrity	O
is	O
compromised	O
,	O
and	O
the	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
-	O
dependent	O
cascade	O
is	O
activated	O
in	O
treated	O
tumor	B-Cell
cells	I-Cell
.	O

However	O
,	O
although	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
and	O
MHC	B-Gene_or_gene_product
class	I-Gene_or_gene_product
I	I-Gene_or_gene_product
are	O
expressed	O
on	O
normal	O
hematopoietic	B-Cell
cells	I-Cell
,	O
which	O
is	O
a	O
potential	O
safety	O
concern	O
,	O
the	O
monoclonal	O
antibodies	O
were	O
selective	O
to	O
tumor	B-Cell
cells	I-Cell
and	O
did	O
not	O
damage	O
normal	O
cells	B-Cell
in	O
vitro	O
or	O
in	O
human	B-Organism
-	O
like	O
mouse	B-Organism
models	O
.	O

These	O
findings	O
suggest	O
that	O
targeting	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
or	O
MHC	B-Gene_or_gene_product
class	I-Gene_or_gene_product
I	I-Gene_or_gene_product
by	O
using	O
antibodies	O
or	O
other	O
agents	O
offers	O
a	O
potential	O
therapeutic	O
approach	O
for	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
MHC	B-Gene_or_gene_product
class	I-Gene_or_gene_product
I	I-Gene_or_gene_product
-	O
expressing	O
malignancies	O
.	O

Cancer	O
2010	O
.	O

(	O
c	O
)	O
2010	O
American	O
Cancer	O
Society	O
.	O

Loss	O
of	O
CDC4	B-Gene_or_gene_product
/	O
FBXW7	B-Gene_or_gene_product
in	O
gastric	B-Cancer
carcinoma	I-Cancer
.	O

BACKGROUND	O
:	O
CDC4	B-Gene_or_gene_product
/	O
FBXW7	B-Gene_or_gene_product
,	O
encoding	O
a	O
ubiquitin	B-Gene_or_gene_product
ligase	O
,	O
maps	O
to	O
4q32	O
and	O
has	O
been	O
implicated	O
as	O
a	O
tumor	B-Cancer
suppressor	O
gene	O
and	O
therapeutic	O
target	O
in	O
many	O
tumor	B-Cancer
types	O
.	O

Mutations	O
in	O
colonic	B-Cancer
adenomas	I-Cancer
,	O
and	O
the	O
frequent	O
losses	O
on	O
4q	B-Cellular_component
described	O
in	O
gastric	B-Cancer
cancer	I-Cancer
prompt	O
speculation	O
about	O
the	O
role	O
of	O
CDC4	B-Gene_or_gene_product
/	O
FBXW7	B-Gene_or_gene_product
in	O
gastric	B-Organ
carcinogenesis	O
.	O

METHODS	O
:	O
We	O
assessed	O
the	O
role	O
of	O
CDC4	B-Gene_or_gene_product
/	O
FBXW7	B-Gene_or_gene_product
in	O
gastric	B-Cancer
cancer	I-Cancer
,	O
through	O
loss	O
of	O
heterozygosity	O
(	O
LOH	O
)	O
and	O
multiplex	O
ligation	O
-	O
dependent	O
probe	O
amplification	O
(	O
MLPA	O
)	O
on	O
47	O
flow	O
-	O
sorted	O
gastric	B-Cancer
carcinomas	I-Cancer
including	O
early	B-Cancer
-	I-Cancer
onset	I-Cancer
gastric	I-Cancer
cancers	I-Cancer
(	O
EOGC	B-Cancer
)	O
and	O
xenografted	O
conventional	O
gastric	B-Cancer
carcinomas	I-Cancer
.	O

Ploidy	O
analysis	O
was	O
carried	O
out	O
on	O
39	O
EOGCs	B-Cancer
and	O
immunohistochemistry	O
of	O
CDC4	B-Gene_or_gene_product
/	O
FBXW7	B-Gene_or_gene_product
and	O
its	O
substrates	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
,	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
jun	I-Gene_or_gene_product
,	O
Notch	B-Gene_or_gene_product
and	O
cyclin	B-Gene_or_gene_product
E	I-Gene_or_gene_product
was	O
performed	O
on	O
204	O
gastric	B-Cancer
carcinomas	I-Cancer
using	O
tissue	B-Tissue
microarrays	O
(	O
TMAs	O
)	O
.	O

Sequence	O
analysis	O
of	O
CDC4	B-Gene_or_gene_product
/	O
FBXW7	B-Gene_or_gene_product
was	O
carried	O
out	O
on	O
gastric	B-Cell
carcinoma	I-Cell
cell	I-Cell
lines	I-Cell
and	O
xenografts	B-Cancer
.	O

RESULTS	O
:	O
Loss	O
of	O
heterozygosity	O
of	O
CDC4	B-Gene_or_gene_product
/	O
FBXW7	B-Gene_or_gene_product
occurred	O
in	O
32	O
%	O
of	O
EOGCs	B-Cancer
,	O
and	O
correlated	O
with	O
loss	O
of	O
expression	O
in	O
26	O
%	O
.	O

Loss	O
of	O
expression	O
was	O
frequent	O
in	O
both	O
EOGC	B-Cancer
and	O
conventional	O
gastric	B-Cancer
cancers	I-Cancer
.	O

No	O
CDC4	B-Gene_or_gene_product
/	O
FBXW7	B-Gene_or_gene_product
mutations	O
were	O
found	O
and	O
loss	O
of	O
CDC4	B-Gene_or_gene_product
/	O
FBXW7	B-Gene_or_gene_product
did	O
not	O
correlate	O
with	O
ploidy	O
status	O
.	O

There	O
was	O
a	O
significant	O
correlation	O
between	O
loss	O
of	O
CDC4	B-Gene_or_gene_product
/	O
FBXW7	B-Gene_or_gene_product
expression	O
and	O
upregulation	O
of	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
.	O

CONCLUSION	O
:	O
Loss	O
of	O
CDC4	B-Gene_or_gene_product
/	O
FBXW7	B-Gene_or_gene_product
appears	O
to	O
play	O
a	O
role	O
in	O
both	O
EOGC	B-Cancer
and	O
conventional	O
gastric	B-Organ
carcinogenesis	O
,	O
and	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
overexpression	O
is	O
likely	O
to	O
be	O
an	O
important	O
oncogenic	O
consequence	O
of	O
CDC4	B-Gene_or_gene_product
/	O
FBXW7	B-Gene_or_gene_product
loss	O
.	O

Angiogenesis	O
inhibitors	O
:	O
current	O
strategies	O
and	O
future	O
prospects	O
.	O

Angiogenesis	O
has	O
become	O
an	O
attractive	O
target	O
for	O
drug	O
therapy	O
because	O
of	O
its	O
key	O
role	O
in	O
tumor	B-Cancer
growth	O
.	O

An	O
extensive	O
array	O
of	O
compounds	O
is	O
currently	O
in	O
preclinical	O
development	O
,	O
with	O
many	O
now	O
entering	O
the	O
clinic	O
and	O
/	O
or	O
achieving	O
approval	O
from	O
the	O
US	O
Food	O
and	O
Drug	O
Administration	O
.	O

Several	O
regulatory	O
and	O
signaling	O
molecules	O
governing	O
angiogenesis	O
are	O
of	O
interest	O
,	O
including	O
growth	O
factors	O
(	O
eg	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
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
fibroblast	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
,	O
and	O
epidermal	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
)	O
,	O
receptor	O
tyrosine	O
kinases	O
,	O
and	O
transcription	O
factors	O
such	O
as	O
hypoxia	B-Gene_or_gene_product
inducible	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
,	O
as	O
well	O
as	O
molecules	O
involved	O
in	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
and	O
phosphoinositide	B-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
signaling	O
.	O

Pharmacologic	O
agents	O
have	O
been	O
identified	O
that	O
target	O
these	O
pathways	O
,	O
yet	O
for	O
some	O
agents	O
(	O
notably	O
thalidomide	B-Simple_chemical
)	O
,	O
an	O
understanding	O
of	O
the	O
specific	O
mechanisms	O
of	O
antitumor	B-Cancer
action	O
has	O
proved	O
elusive	O
.	O

The	O
following	O
review	O
describes	O
key	O
molecular	O
mechanisms	O
and	O
novel	O
therapies	O
that	O
are	O
on	O
the	O
horizon	O
for	O
antiangiogenic	O
tumor	B-Cancer
therapy	O
.	O

Overexpression	O
of	O
Bax	B-Gene_or_gene_product
inhibitor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
BI	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
induces	O
cell	B-Cell
transformation	O
in	O
NIH3T3	B-Cell
cells	I-Cell
.	O

BI	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
Bax	B-Gene_or_gene_product
inhibitor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
,	O
an	O
apoptosis	O
-	O
inhibiting	O
gene	O
belonging	O
to	O
the	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
protein	O
family	O
,	O
plays	O
an	O
important	O
role	O
in	O
mitochondrial	B-Cellular_component
apoptosis	O
pathway	O
to	O
suppress	O
Bax	B-Gene_or_gene_product
-	O
induced	O
apoptosis	O
.	O

To	O
investigate	O
the	O
potential	O
role	O
of	O
BI	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
in	O
promoting	O
cell	B-Cell
growth	O
and	O
tumorigenesis	O
,	O
in	O
the	O
present	O
study	O
we	O
overexpressed	O
the	O
BI	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
gene	O
in	O
NIH3T3	B-Cell
cells	I-Cell
using	O
the	O
lentivirus	B-Organism
-	O
mediated	O
gene	O
expression	O
system	O
.	O

Our	O
in	O
vitro	O
studies	O
showed	O
that	O
NIH3T3	B-Cell
cells	I-Cell
overexpressing	O
BI	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
displayed	O
a	O
significantly	O
higher	O
growth	O
rate	O
and	O
formed	O
more	O
and	O
larger	O
colonies	O
than	O
the	O
control	B-Cell
cells	I-Cell
.	O

In	O
addition	O
,	O
our	O
in	O
vivo	O
studies	O
indicated	O
that	O
the	O
lenti	O
-	O
BI	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
infected	O
cells	B-Cell
formed	O
obvious	O
tumours	B-Cancer
,	O
while	O
no	O
tumours	B-Cancer
were	O
formed	O
by	O
the	O
control	B-Cell
cells	I-Cell
after	O
subcutaneously	O
injected	O
into	O
nude	B-Organism
mice	I-Organism
.	O

These	O
results	O
strongly	O
suggested	O
that	O
the	O
BI	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
gene	O
might	O
play	O
a	O
crucial	O
role	O
in	O
neoplastic	O
genesis	O
and	O
development	O
.	O

Anti	O
-	O
metastasis	O
effects	O
of	O
gallic	B-Simple_chemical
acid	I-Simple_chemical
on	O
gastric	B-Cell
cancer	I-Cell
cells	I-Cell
involves	O
inhibition	O
of	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
activity	O
and	O
downregulation	O
of	O
PI3K	B-Gene_or_gene_product
/	O
AKT	B-Gene_or_gene_product
/	O
small	B-Gene_or_gene_product
GTPase	I-Gene_or_gene_product
signals	O
.	O

Polyphenols	B-Simple_chemical
are	O
natural	O
antioxidants	O
that	O
are	O
thought	O
to	O
contribute	O
to	O
prevention	O
of	O
cardiovascular	B-Anatomical_system
disease	O
and	O
malignancy	O
.	O

Although	O
many	O
studies	O
have	O
been	O
carried	O
out	O
to	O
investigate	O
the	O
chemopreventive	O
role	O
of	O
flavonoids	B-Simple_chemical
,	O
less	O
attention	O
has	O
been	O
focused	O
on	O
phenolic	B-Simple_chemical
acids	I-Simple_chemical
.	O

In	O
this	O
study	O
,	O
the	O
aim	O
was	O
to	O
investigate	O
the	O
effect	O
of	O
phenolic	B-Simple_chemical
acids	I-Simple_chemical
found	O
abundantly	O
in	O
vegetables	B-Organism_subdivision
,	O
i	O
.	O
e	O
.	O
gallic	B-Simple_chemical
acid	I-Simple_chemical
(	O
GA	B-Simple_chemical
)	O
,	O
caffeic	B-Simple_chemical
acid	I-Simple_chemical
(	O
CA	B-Simple_chemical
)	O
and	O
protocatechuic	B-Simple_chemical
acid	I-Simple_chemical
(	O
PCA	B-Simple_chemical
)	O
,	O
on	O
the	O
inhibition	O
of	O
gastric	B-Cell
adenocarcinoma	I-Cell
(	I-Cell
AGS	I-Cell
)	I-Cell
cell	I-Cell
metastasis	O
.	O

The	O
results	O
showed	O
0	O
.	O
01	O
mM	O
GA	B-Simple_chemical
induced	O
the	O
same	O
level	O
of	O
cell	B-Cell
toxicity	O
as	O
4	O
.	O
0mM	O
PCA	B-Simple_chemical
.	O

Using	O
wound	B-Pathological_formation
-	O
healing	O
assay	O
and	O
Boyden	O
chamber	O
assay	O
,	O
GA	B-Simple_chemical
had	O
potent	O
inhibitory	O
effects	O
on	O
AGS	B-Cell
cell	I-Cell
migration	O
.	O

The	O
expression	O
of	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
/	O
9	B-Gene_or_gene_product
of	O
AGS	B-Cell
cells	I-Cell
was	O
inhibited	O
by	O
2	O
.	O
0	O
microM	O
of	O
GA	B-Simple_chemical
.	O

It	O
is	O
possible	O
that	O
the	O
suppressive	O
effect	O
of	O
GA	B-Simple_chemical
on	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
/	O
9	B-Gene_or_gene_product
might	O
involve	O
the	O
inhibition	O
of	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
activity	O
.	O

Multiple	O
proteins	O
involved	O
in	O
metastasis	O
and	O
the	O
cytoskeletal	B-Cellular_component
reorganization	O
signal	O
pathway	O
,	O
including	O
Ras	B-Gene_or_gene_product
,	O
Cdc42	B-Gene_or_gene_product
,	O
Rac1	B-Gene_or_gene_product
,	O
RhoA	B-Gene_or_gene_product
,	O
RhoB	B-Gene_or_gene_product
,	O
PI3K	B-Gene_or_gene_product
and	O
p38MAPK	B-Gene_or_gene_product
,	O
were	O
also	O
inhibited	O
by	O
GA	B-Simple_chemical
.	O

Furthermore	O
,	O
immunoreactivity	O
assay	O
of	O
cytoskeletal	B-Cellular_component
F	B-Gene_or_gene_product
-	I-Gene_or_gene_product
actin	I-Gene_or_gene_product
demonstrated	O
a	O
significant	O
inhibitory	O
effect	O
of	O
GA	B-Simple_chemical
treatment	O
.	O

In	O
conclusion	O
,	O
GA	B-Simple_chemical
may	O
have	O
the	O
potential	O
to	O
be	O
an	O
effective	O
agent	O
for	O
prevention	O
and	O
treatment	O
of	O
gastric	B-Cancer
cancer	I-Cancer
metastasis	O
.	O

1	B-Simple_chemical
,	I-Simple_chemical
4	I-Simple_chemical
-	I-Simple_chemical
phenylenebis	I-Simple_chemical
(	I-Simple_chemical
methylene	I-Simple_chemical
)	I-Simple_chemical
selenocyanate	I-Simple_chemical
,	O
but	O
not	O
selenomethionine	B-Simple_chemical
,	O
inhibits	O
androgen	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
and	O
Akt	B-Gene_or_gene_product
signaling	O
in	O
human	B-Organism
prostate	B-Cell
cancer	I-Cell
cells	I-Cell
.	O

The	O
lack	O
of	O
treatment	O
for	O
worried	O
-	O
well	O
patients	B-Organism
with	O
high	O
-	O
grade	O
prostatic	B-Pathological_formation
intraepithelial	I-Pathological_formation
neoplasia	I-Pathological_formation
combined	O
with	O
issues	O
of	O
recurrence	O
and	O
hormone	O
resistance	O
in	O
prostate	B-Cancer
cancer	I-Cancer
survivors	O
remains	O
a	O
major	O
public	O
health	O
obstacle	O
.	O

The	O
long	O
latency	O
of	O
prostate	B-Cancer
cancer	I-Cancer
development	O
provides	O
an	O
opportunity	O
to	O
intervene	O
with	O
agents	O
of	O
known	O
mechanisms	O
at	O
various	O
stages	O
of	O
disease	O
progression	O
.	O

A	O
number	O
of	O
signaling	O
cascades	O
have	O
been	O
shown	O
to	O
play	O
important	O
roles	O
in	O
prostate	B-Cancer
cancer	I-Cancer
development	O
and	O
progression	O
,	O
including	O
the	O
androgen	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
(	O
AR	B-Gene_or_gene_product
)	O
and	O
phosphatidylinositol	B-Gene_or_gene_product
3	I-Gene_or_gene_product
-	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
/	O
Akt	B-Gene_or_gene_product
signaling	O
pathways	O
.	O

Crosstalk	O
between	O
these	O
two	O
pathways	O
is	O
also	O
thought	O
to	O
contribute	O
to	O
progression	O
and	O
hormone	O
-	O
refractory	O
prostate	B-Cancer
disease	I-Cancer
.	O

Our	O
initial	O
investigations	O
show	O
that	O
the	O
naturally	O
occurring	O
organoselenium	O
compound	O
selenomethionine	B-Simple_chemical
(	O
SM	B-Simple_chemical
)	O
and	O
the	O
synthetic	O
1	B-Simple_chemical
,	I-Simple_chemical
4	I-Simple_chemical
-	I-Simple_chemical
phenylenebis	I-Simple_chemical
(	I-Simple_chemical
methylene	I-Simple_chemical
)	I-Simple_chemical
selenocyanate	I-Simple_chemical
(	O
p	B-Simple_chemical
-	I-Simple_chemical
XSC	I-Simple_chemical
)	O
can	O
inhibit	O
human	B-Organism
prostate	B-Cell
cancer	I-Cell
cell	I-Cell
viability	O
;	O
however	O
,	O
in	O
contrast	O
to	O
SM	B-Simple_chemical
,	O
p	B-Simple_chemical
-	I-Simple_chemical
XSC	I-Simple_chemical
is	O
active	O
at	O
physiologically	O
relevant	O
doses	O
.	O

In	O
the	O
current	O
investigation	O
,	O
we	O
show	O
that	O
p	B-Simple_chemical
-	I-Simple_chemical
XSC	I-Simple_chemical
,	O
but	O
not	O
an	O
equivalent	O
dose	O
of	O
SM	B-Simple_chemical
,	O
alters	O
molecular	O
targets	O
and	O
induces	O
apoptosis	O
in	O
androgen	B-Simple_chemical
-	O
responsive	O
LNCaP	B-Cell
and	O
androgen	B-Simple_chemical
-	O
independent	O
LNCaP	B-Cell
C4	I-Cell
-	I-Cell
2	I-Cell
human	I-Cell
prostate	I-Cell
cancer	I-Cell
cells	I-Cell
.	O

p	B-Simple_chemical
-	I-Simple_chemical
XSC	I-Simple_chemical
effectively	O
inhibits	O
AR	B-Gene_or_gene_product
expression	O
and	O
transcriptional	O
activity	O
in	O
both	O
cell	B-Cell
lines	I-Cell
.	O

p	B-Simple_chemical
-	I-Simple_chemical
XSC	I-Simple_chemical
also	O
decreases	O
Akt	B-Gene_or_gene_product
phosphorylation	O
as	O
well	O
as	O
Akt	B-Gene_or_gene_product
-	O
specific	O
phosphorylation	O
of	O
the	O
AR	B-Gene_or_gene_product
.	O

Inhibition	O
of	O
Akt	B-Gene_or_gene_product
,	O
however	O
,	O
does	O
not	O
fully	O
attenuate	O
p	B-Simple_chemical
-	I-Simple_chemical
XSC	I-Simple_chemical
-	O
mediated	O
downregulation	O
of	O
AR	B-Gene_or_gene_product
activity	O
,	O
suggesting	O
that	O
inhibition	O
of	O
AR	B-Gene_or_gene_product
signaling	O
by	O
p	B-Simple_chemical
-	I-Simple_chemical
XSC	I-Simple_chemical
does	O
not	O
occur	O
solely	O
through	O
alterations	O
in	O
the	O
phosphatidylinositol	B-Gene_or_gene_product
3	I-Gene_or_gene_product
-	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
/	O
Akt	B-Gene_or_gene_product
survival	O
pathway	O
.	O

Our	O
data	O
suggest	O
that	O
p	B-Simple_chemical
-	I-Simple_chemical
XSC	I-Simple_chemical
inhibits	O
multiple	O
signaling	O
pathways	O
in	O
prostate	B-Cancer
cancer	I-Cancer
,	O
likely	O
accounting	O
for	O
the	O
downstream	O
effects	O
on	O
proliferation	O
and	O
apoptosis	O
.	O

Sanguinarine	B-Simple_chemical
induces	O
apoptosis	O
of	O
human	B-Organism
osteosarcoma	B-Cell
cells	I-Cell
through	O
the	O
extrinsic	O
and	O
intrinsic	O
pathways	O
.	O

The	O
quaternary	O
benzo	B-Simple_chemical
[	I-Simple_chemical
c	I-Simple_chemical
]	I-Simple_chemical
phenanthridine	I-Simple_chemical
alkaloid	I-Simple_chemical
sanguinarine	B-Simple_chemical
inhibits	O
the	O
proliferation	O
of	O
cancerous	B-Cell
cells	I-Cell
from	O
different	O
origins	O
,	O
including	O
lung	B-Organ
,	O
breast	B-Organism_subdivision
,	O
pancreatic	B-Organ
and	O
colon	B-Organ
,	O
but	O
nothing	O
is	O
known	O
of	O
its	O
effects	O
on	O
osteosarcoma	B-Cancer
,	O
a	O
primary	B-Cancer
malignant	I-Cancer
bone	I-Cancer
tumour	I-Cancer
.	O

We	O
have	O
found	O
that	O
sanguinarine	B-Simple_chemical
alters	O
the	O
morphology	O
and	O
reduces	O
the	O
viability	O
of	O
MG	B-Cell
-	I-Cell
63	I-Cell
and	O
SaOS	B-Cell
-	I-Cell
2	I-Cell
human	I-Cell
osteosarcoma	I-Cell
cell	I-Cell
lines	I-Cell
in	O
concentration	O
-	O
and	O
time	O
-	O
dependent	O
manner	O
.	O

Incubation	O
with	O
1	O
micromol	O
/	O
L	O
sanguinarine	B-Simple_chemical
for	O
4	O
and	O
24h	O
killed	O
more	O
efficiently	O
MG	B-Cell
-	I-Cell
63	I-Cell
cells	I-Cell
than	O
SaOS	B-Cell
-	I-Cell
2	I-Cell
cells	I-Cell
,	O
while	O
incubation	O
with	O
5	O
micromol	O
/	O
L	O
sanguinarine	B-Simple_chemical
killed	O
almost	O
100	O
%	O
of	O
both	O
cell	B-Cell
populations	I-Cell
within	O
24h	O
.	O

This	O
treatment	O
also	O
changed	O
the	O
mitochondrial	B-Cellular_component
membrane	I-Cellular_component
potential	O
in	O
both	O
MG	B-Cell
-	I-Cell
63	I-Cell
and	O
SaOS	B-Cell
-	I-Cell
2	I-Cell
cells	I-Cell
within	O
1h	O
,	O
caused	O
chromatin	B-Cellular_component
condensation	O
and	O
the	O
formation	O
of	O
apoptotic	B-Cellular_component
bodies	I-Cellular_component
.	O

It	O
activated	O
multicaspases	O
,	O
and	O
increased	O
the	O
activities	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
9	I-Gene_or_gene_product
in	O
both	O
MG	B-Cell
-	I-Cell
63	I-Cell
and	O
SaOS	B-Cell
-	I-Cell
2	I-Cell
cells	I-Cell
.	O

These	O
data	O
highlight	O
sanguinarine	B-Simple_chemical
as	O
a	O
novel	O
potential	O
agent	O
for	O
bone	B-Cancer
cancer	I-Cancer
therapy	O
.	O

Actin	B-Gene_or_gene_product
-	O
sequestering	O
protein	O
,	O
thymosin	B-Gene_or_gene_product
beta	I-Gene_or_gene_product
-	I-Gene_or_gene_product
4	I-Gene_or_gene_product
,	O
is	O
a	O
novel	O
hypoxia	O
responsive	O
regulator	O
.	O

Angiogenesis	O
is	O
induced	O
by	O
soluble	O
factors	O
such	O
as	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
released	O
from	O
tumor	B-Cell
cells	I-Cell
in	O
hypoxia	O
.	O

It	O
enhances	O
solid	B-Cancer
tumor	I-Cancer
growth	O
and	O
provides	O
an	O
ability	O
to	O
establish	O
metastasis	O
at	O
peripheral	B-Multi-tissue_structure
sites	I-Multi-tissue_structure
by	O
tumor	B-Cell
cell	I-Cell
migration	O
.	O

Thymosin	B-Gene_or_gene_product
beta	I-Gene_or_gene_product
-	I-Gene_or_gene_product
4	I-Gene_or_gene_product
(	O
TB4	B-Gene_or_gene_product
)	O
is	O
an	O
actin	B-Gene_or_gene_product
-	O
sequestering	O
protein	O
to	O
control	O
cytoskeletal	B-Cellular_component
reorganization	O
.	O

Here	O
,	O
we	O
investigated	O
whether	O
angiogenesis	O
and	O
tumor	B-Cancer
metastasis	O
are	O
dependent	O
on	O
hypoxia	O
conditioning	O
-	O
induced	O
TB4	B-Gene_or_gene_product
expression	O
in	O
B16F10	B-Cell
melanoma	I-Cell
cells	I-Cell
.	O

TB4	B-Gene_or_gene_product
expression	O
in	O
B16F10	B-Cell
cells	I-Cell
was	O
increased	O
by	O
hypoxia	O
conditioning	O
in	O
a	O
time	O
-	O
dependent	O
manner	O
.	O

In	O
addition	O
,	O
we	O
found	O
an	O
increase	O
of	O
angiogenesis	O
and	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
expression	O
in	O
TB4	B-Gene_or_gene_product
-	O
transgenic	O
(	O
Tg	O
)	O
mice	O
as	O
compared	O
to	O
wildtype	O
mice	B-Organism
.	O

When	O
wound	B-Pathological_formation
healing	O
assay	O
was	O
used	O
to	O
assess	O
in	O
vitro	O
tumor	B-Cell
cell	I-Cell
migration	O
,	O
hypoxia	O
conditioning	O
for	O
1	O
h	O
enhanced	O
B16F10	B-Cell
cell	I-Cell
migration	O
.	O

When	O
TB4	B-Gene_or_gene_product
expression	O
in	O
B16F10	B-Cell
cells	I-Cell
was	O
inhibited	O
by	O
the	O
infection	O
with	O
small	O
hairpin	O
(	O
sh	O
)	O
RNA	O
of	O
TB4	B-Gene_or_gene_product
cloned	O
in	O
lentiviral	B-Organism
vector	O
,	O
tumor	B-Cell
cell	I-Cell
migration	O
was	O
retarded	O
.	O

In	O
addition	O
,	O
hypoxia	O
conditioning	O
-	O
induced	O
tumor	B-Cell
cell	I-Cell
migration	O
was	O
reduced	O
by	O
the	O
infection	O
of	O
lentiviral	B-Organism
shRNA	O
of	O
TB4	B-Gene_or_gene_product
.	O

HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
stabilization	O
and	O
the	O
expression	O
of	O
VEGF	B-Gene_or_gene_product
isoform	O
165	O
and	O
121	O
in	O
hypoxia	O
were	O
also	O
reduced	O
by	O
the	O
infection	O
of	O
lentiviral	B-Organism
shRNA	O
of	O
TB4	B-Gene_or_gene_product
in	O
B16F10	B-Cell
cells	I-Cell
.	O

We	O
also	O
found	O
an	O
increase	O
of	O
tumor	B-Cancer
growth	O
and	O
lung	B-Organ
metastasis	O
count	O
in	O
TB4	B-Gene_or_gene_product
-	O
Tg	O
mice	O
as	O
compared	O
to	O
wildtype	O
mice	B-Organism
.	O

Collectively	O
,	O
hypoxia	O
conditioning	O
induced	O
tumor	B-Cell
cell	I-Cell
migration	O
by	O
TB4	B-Gene_or_gene_product
expression	O
-	O
dependent	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
stabilization	O
.	O

It	O
suggests	O
that	O
TB4	B-Gene_or_gene_product
could	O
be	O
a	O
hypoxia	O
responsive	O
regulator	O
to	O
control	O
tumor	B-Cell
cell	I-Cell
migration	O
in	O
angiogenesis	O
and	O
tumor	B-Cancer
metastasis	O
.	O

Androgen	B-Simple_chemical
stimulates	O
glycolysis	O
for	O
de	O
novo	O
lipid	B-Simple_chemical
synthesis	O
by	O
increasing	O
the	O
activities	O
of	O
hexokinase	B-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
6	B-Gene_or_gene_product
-	I-Gene_or_gene_product
phosphofructo	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
-	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
/	I-Gene_or_gene_product
fructose	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	I-Gene_or_gene_product
6	I-Gene_or_gene_product
-	I-Gene_or_gene_product
bisphosphatase	I-Gene_or_gene_product
2	I-Gene_or_gene_product
in	O
prostate	B-Cell
cancer	I-Cell
cells	I-Cell
.	O

Up	O
-	O
regulation	O
of	O
lipogenesis	O
by	O
androgen	B-Simple_chemical
is	O
one	O
of	O
the	O
most	O
characteristic	O
metabolic	O
features	O
of	O
LNCaP	B-Cell
prostate	I-Cell
cancer	I-Cell
cells	I-Cell
.	O

The	O
present	O
study	O
revealed	O
that	O
androgen	B-Simple_chemical
increases	O
glucose	B-Simple_chemical
utilization	O
for	O
de	O
novo	O
lipogenesis	O
in	O
LNCaP	B-Cell
cells	I-Cell
through	O
the	O
activation	O
of	O
HK2	B-Gene_or_gene_product
(	O
hexokinase	B-Gene_or_gene_product
2	I-Gene_or_gene_product
)	I-Gene_or_gene_product
and	O
activation	O
of	O
the	O
cardiac	B-Organ
isoform	O
of	O
PFKFB2	B-Gene_or_gene_product
(	O
6	B-Gene_or_gene_product
-	I-Gene_or_gene_product
phosphofructo	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
-	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
/	I-Gene_or_gene_product
fructose	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	I-Gene_or_gene_product
6	I-Gene_or_gene_product
-	I-Gene_or_gene_product
bisphosphatase	I-Gene_or_gene_product
)	O
.	O

Activation	O
of	O
PKA	B-Gene_or_gene_product
(	O
cAMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
dependent	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
)	O
by	O
androgen	B-Simple_chemical
increased	O
phosphorylation	O
of	O
CREB	B-Gene_or_gene_product
[	O
CRE	B-Gene_or_gene_product
(	I-Gene_or_gene_product
cAMP	I-Gene_or_gene_product
-	I-Gene_or_gene_product
response	I-Gene_or_gene_product
element	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	I-Gene_or_gene_product
binding	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
]	O
,	O
which	O
in	O
turn	O
bound	O
to	O
CRE	O
on	O
the	O
promoter	O
of	O
the	O
HK2	B-Gene_or_gene_product
gene	O
resulting	O
in	O
transcriptional	O
activation	O
of	O
the	O
HK2	B-Gene_or_gene_product
gene	O
.	O

Up	O
-	O
regulation	O
of	O
PFKFB2	B-Gene_or_gene_product
expression	O
was	O
mediated	O
by	O
the	O
direct	O
binding	O
of	O
ligand	O
-	O
activated	O
androgen	B-Simple_chemical
receptor	O
to	O
the	O
PFKFB2	B-Gene_or_gene_product
promoter	O
.	O

The	O
activated	O
PI3K	B-Gene_or_gene_product
(	O
phosphoinositide	B-Gene_or_gene_product
3	I-Gene_or_gene_product
-	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
)	O
/	O
Akt	B-Gene_or_gene_product
signalling	O
pathway	O
in	O
LNCaP	B-Cell
cells	I-Cell
contributes	O
to	O
the	O
phosphorylation	O
of	O
PFKFB2	B-Gene_or_gene_product
at	O
Ser466	B-Amino_acid
and	O
Ser483	B-Amino_acid
,	O
resulting	O
in	O
the	O
constitutive	O
activation	O
of	O
PFK	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
(	O
6	B-Gene_or_gene_product
-	I-Gene_or_gene_product
phosphofructo	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
-	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
)	O
activity	O
.	O

Glucose	B-Simple_chemical
uptake	O
and	O
lipogenesis	O
were	O
severely	O
blocked	O
by	O
knocking	O
-	O
down	O
of	O
PFKFB2	B-Gene_or_gene_product
using	O
siRNA	O
(	O
small	O
interfering	O
RNA	O
)	O
or	O
by	O
inhibition	O
of	O
PFK	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
activity	O
with	O
LY294002	B-Simple_chemical
treatment	O
.	O

Taken	O
together	O
,	O
our	O
results	O
suggest	O
that	O
the	O
induction	O
of	O
de	O
novo	O
lipid	B-Simple_chemical
synthesis	O
by	O
androgen	B-Simple_chemical
requires	O
the	O
transcriptional	O
up	O
-	O
regulation	O
of	O
HK2	B-Gene_or_gene_product
and	O
PFKFB2	B-Gene_or_gene_product
,	O
and	O
phosphorylation	O
of	O
PFKFB2	B-Gene_or_gene_product
generated	O
by	O
the	O
PI3K	B-Gene_or_gene_product
/	O
Akt	B-Gene_or_gene_product
signalling	O
pathway	O
to	O
supply	O
the	O
source	O
for	O
lipogenesis	O
from	O
glucose	B-Simple_chemical
in	O
prostate	B-Cell
cancer	I-Cell
cells	I-Cell
.	O

Diallyl	B-Simple_chemical
trisulfide	I-Simple_chemical
(	O
DATS	B-Simple_chemical
)	O
inhibits	O
mouse	B-Organism
colon	B-Cancer
tumor	I-Cancer
in	O
mouse	B-Organism
CT	B-Cell
-	I-Cell
26	I-Cell
cells	I-Cell
allograft	O
model	O
in	O
vivo	O
.	O

Our	O
earlier	O
studies	O
showed	O
that	O
DATS	B-Simple_chemical
induced	O
apoptosis	O
in	O
human	B-Organism
colon	B-Cell
cancer	I-Cell
HT29	I-Cell
and	O
colo	B-Cell
205	I-Cell
cell	I-Cell
lines	I-Cell
in	O
vitro	O
.	O

However	O
,	O
there	O
is	O
no	O
report	O
to	O
show	O
that	O
DATS	B-Simple_chemical
induced	O
apoptosis	O
in	O
vitro	O
and	O
inhibited	O
CT26	B-Cell
cancer	I-Cell
cells	I-Cell
in	O
vivo	O
on	O
a	O
murine	B-Organism
allograft	O
animal	O
model	O
.	O

In	O
vitro	O
studies	O
,	O
the	O
results	O
indicated	O
that	O
DATS	B-Simple_chemical
induced	O
morphological	O
changes	O
and	O
induction	O
of	O
apoptosis	O
in	O
CT26	B-Cell
cells	I-Cell
.	O

In	O
vivo	O
studies	O
,	O
CT26	B-Cell
cancer	I-Cell
cells	I-Cell
were	O
implanted	O
into	O
BALB	B-Organism
/	I-Organism
c	I-Organism
mice	I-Organism
and	O
groups	O
of	O
mice	B-Organism
were	O
treated	O
with	O
vehicle	O
,	O
DATS	B-Simple_chemical
(	O
10	O
and	O
50	O
mg	O
/	O
kg	O
of	O
body	B-Organism_subdivision
weight	O
)	O
.	O

DATS	B-Simple_chemical
were	O
injected	O
once	O
per	O
four	O
days	O
intraperitoneally	B-Immaterial_anatomical_entity
(	O
i	O
.	O
p	O
.	O
)	O
,	O
with	O
treatment	O
starting	O
4	O
weeks	O
prior	O
to	O
cells	B-Cell
inoculation	O
.	O

Treatment	O
with	O
vehicle	O
or	O
with	O
10	O
and	O
50	O
mg	O
/	O
kg	O
of	O
DATS	B-Simple_chemical
resulted	O
in	O
a	O
reduction	O
in	O
tumor	B-Cancer
volume	O
and	O
weight	O
.	O

Tumor	B-Cancer
volume	O
and	O
total	O
hemoglobin	B-Gene_or_gene_product
in	O
allograft	O
mice	B-Organism
treated	O
with	O
50	O
mg	O
/	O
kg	O
DATS	B-Simple_chemical
were	O
significantly	O
smaller	O
than	O
that	O
in	O
the	O
control	O
group	O
.	O

These	O
findings	O
indicated	O
that	O
DATS	B-Simple_chemical
inhibits	O
tumor	B-Cancer
growth	O
in	O
an	O
allograft	O
animal	O
model	O
.	O

Thus	O
,	O
DATS	B-Simple_chemical
may	O
represent	O
a	O
colon	B-Cancer
cancer	I-Cancer
preventive	O
agent	O
and	O
can	O
be	O
used	O
in	O
the	O
future	O
.	O

High	B-Gene_or_gene_product
-	I-Gene_or_gene_product
mobility	I-Gene_or_gene_product
group	I-Gene_or_gene_product
A2	I-Gene_or_gene_product
protein	O
modulates	O
hTERT	B-Gene_or_gene_product
transcription	O
to	O
promote	O
tumorigenesis	O
.	O

The	O
high	B-Gene_or_gene_product
-	I-Gene_or_gene_product
mobility	I-Gene_or_gene_product
group	I-Gene_or_gene_product
A2	I-Gene_or_gene_product
gene	O
(	O
HMGA2	B-Gene_or_gene_product
)	O
is	O
one	O
of	O
the	O
most	O
frequently	O
amplified	O
genes	O
in	O
human	B-Organism
cancers	B-Cancer
.	O

However	O
,	O
functions	O
of	O
HMGA2	B-Gene_or_gene_product
in	O
tumorigenesis	O
are	O
not	O
fully	O
understood	O
due	O
to	O
limited	O
knowledge	O
of	O
its	O
targets	O
in	O
tumor	B-Cell
cells	I-Cell
.	O

Our	O
study	O
reveals	O
a	O
novel	O
link	O
between	O
HMGA2	B-Gene_or_gene_product
and	O
the	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
(	O
hTERT	B-Gene_or_gene_product
)	O
,	O
the	O
catalytic	O
subunit	O
of	O
telomerase	B-Gene_or_gene_product
,	O
which	O
offers	O
critical	O
insight	O
into	O
how	O
HMGA2	B-Gene_or_gene_product
contributes	O
to	O
tumorigenesis	O
.	O

The	O
expression	O
of	O
HMGA2	B-Gene_or_gene_product
modulates	O
the	O
expression	O
of	O
hTERT	B-Gene_or_gene_product
,	O
resulting	O
in	O
cells	B-Cell
with	O
enhanced	O
telomerase	B-Gene_or_gene_product
activities	O
and	O
increased	O
telomere	B-Cellular_component
length	O
.	O

Treatment	O
with	O
suberoylanilide	B-Simple_chemical
hydroxamide	I-Simple_chemical
(	O
SAHA	B-Simple_chemical
)	O
,	O
a	O
histone	B-Gene_or_gene_product
deacetylase	I-Gene_or_gene_product
(	O
HDAC	B-Gene_or_gene_product
)	O
inhibitor	O
,	O
causes	O
dose	O
-	O
dependent	O
hTERT	B-Gene_or_gene_product
reporter	O
activation	O
,	O
mimicking	O
HMGA2	B-Gene_or_gene_product
overexpression	O
.	O

By	O
interacting	O
with	O
Sp1	B-Gene_or_gene_product
,	O
HMGA2	B-Gene_or_gene_product
interferes	O
with	O
the	O
recruitment	O
of	O
HDAC2	B-Gene_or_gene_product
to	O
the	O
hTERT	B-Gene_or_gene_product
proximal	O
promoter	O
,	O
enhancing	O
localized	O
histone	B-Gene_or_gene_product
H3	I-Gene_or_gene_product
-	O
K9	O
acetylation	O
and	O
thereby	O
stimulating	O
hTERT	B-Gene_or_gene_product
expression	O
and	O
telomerase	B-Gene_or_gene_product
activity	O
.	O

Moreover	O
,	O
HMGA2	B-Gene_or_gene_product
knockdown	O
by	O
short	O
hairpin	O
HMGA2	B-Gene_or_gene_product
in	O
HepG2	B-Cell
cells	I-Cell
leads	O
to	O
progressive	O
telomere	B-Cellular_component
shortening	O
and	O
a	O
concurrent	O
decrease	O
of	O
steady	O
-	O
state	O
hTERT	B-Gene_or_gene_product
mRNA	O
levels	O
,	O
attenuating	O
their	O
ability	O
to	O
form	O
colonies	O
in	O
soft	O
agar	O
.	O

Importantly	O
,	O
HMGA2	B-Gene_or_gene_product
partially	O
replaces	O
the	O
function	O
of	O
hTERT	B-Gene_or_gene_product
during	O
the	O
tumorigenic	O
transformation	O
of	O
normal	O
human	B-Organism
fibroblasts	B-Cell
.	O

These	O
findings	O
are	O
potentially	O
clinically	O
relevant	O
,	O
because	O
HMGA2	B-Gene_or_gene_product
expression	O
is	O
reported	O
to	O
be	O
upregulated	O
in	O
a	O
number	O
of	O
human	B-Organism
cancers	B-Cancer
as	O
telomere	B-Cellular_component
maintenance	O
is	O
essential	O
for	O
tumorigenesis	O
.	O

Hedgehog	B-Gene_or_gene_product
signaling	O
:	O
networking	O
to	O
nurture	O
a	O
promalignant	B-Cancer
tumor	I-Cancer
microenvironment	O
.	O

In	O
addition	O
to	O
its	O
role	O
in	O
embryonic	B-Developing_anatomical_structure
development	O
,	O
the	O
Hedgehog	B-Gene_or_gene_product
pathway	O
has	O
been	O
shown	O
to	O
be	O
an	O
active	O
participant	O
in	O
cancer	B-Cancer
development	O
,	O
progression	O
,	O
and	O
metastasis	O
.	O

Although	O
this	O
pathway	O
is	O
activated	O
by	O
autocrine	O
signaling	O
by	O
Hedgehog	B-Gene_or_gene_product
ligands	O
,	O
it	O
can	O
also	O
initiate	O
paracrine	O
signaling	O
with	O
cells	B-Cell
in	O
the	O
microenvironment	O
.	O

This	O
creates	O
a	O
network	O
of	O
Hedgehog	B-Gene_or_gene_product
signaling	O
that	O
determines	O
the	O
malignant	O
behavior	O
of	O
the	O
tumor	B-Cell
cells	I-Cell
.	O

As	O
a	O
result	O
of	O
paracrine	O
signal	O
transmission	O
,	O
the	O
effects	O
of	O
Hedgehog	B-Gene_or_gene_product
signaling	O
most	O
profoundly	O
influence	O
the	O
stromal	B-Cell
cells	I-Cell
that	O
constitute	O
the	O
tumor	B-Cancer
microenvironment	O
.	O

The	O
stromal	B-Cell
cells	I-Cell
in	O
turn	O
produce	O
factors	O
that	O
nurture	O
the	O
tumor	B-Cancer
.	O

Thus	O
,	O
such	O
a	O
resonating	O
cross	O
-	O
talk	O
can	O
amplify	O
Hedgehog	B-Gene_or_gene_product
signaling	O
,	O
resulting	O
in	O
molecular	O
chatter	O
that	O
overall	O
promotes	O
tumor	B-Cancer
progression	O
.	O

Inhibitors	O
of	O
Hedgehog	B-Gene_or_gene_product
signaling	O
have	O
been	O
the	O
subject	O
of	O
intense	O
research	O
.	O

Several	O
of	O
these	O
inhibitors	O
are	O
currently	O
being	O
evaluated	O
in	O
clinical	O
trials	O
.	O

Here	O
,	O
we	O
review	O
the	O
role	O
of	O
the	O
Hedgehog	B-Gene_or_gene_product
pathway	O
in	O
the	O
signature	O
characteristics	O
of	O
cancer	B-Cell
cells	I-Cell
that	O
determine	O
tumor	B-Cancer
development	O
,	O
progression	O
,	O
and	O
metastasis	O
.	O

This	O
review	O
condenses	O
the	O
latest	O
findings	O
on	O
the	O
signaling	O
pathways	O
that	O
are	O
activated	O
and	O
/	O
or	O
regulated	O
by	O
molecules	O
generated	O
from	O
Hedgehog	B-Gene_or_gene_product
signaling	O
in	O
cancer	B-Cancer
and	O
cites	O
promising	O
clinical	O
interventions	O
.	O

Finally	O
,	O
we	O
discuss	O
future	O
directions	O
for	O
identifying	O
the	O
appropriate	O
patients	B-Organism
for	O
therapy	O
,	O
developing	O
reliable	O
markers	O
of	O
efficacy	O
of	O
treatment	O
,	O
and	O
combating	O
resistance	O
to	O
Hedgehog	B-Gene_or_gene_product
pathway	O
inhibitors	O
.	O

Analysis	O
of	O
molecular	O
aberrations	O
of	O
Wnt	B-Gene_or_gene_product
pathway	O
gladiators	O
in	O
colorectal	B-Cancer
cancer	I-Cancer
in	O
the	O
Kashmiri	O
population	O
.	O

The	O
development	O
and	O
progression	O
of	O
colorectal	B-Cancer
cancer	I-Cancer
(	O
CRC	B-Cancer
)	O
is	O
a	O
multi	O
-	O
step	O
process	O
,	O
and	O
the	O
Wnt	B-Gene_or_gene_product
pathways	O
with	O
its	O
two	O
molecular	O
gladiators	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
and	O
beta	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
plays	O
an	O
important	O
role	O
in	O
transforming	O
a	O
normal	B-Tissue
tissue	I-Tissue
into	O
a	O
malignant	B-Cancer
one	I-Cancer
.	O

In	O
this	O
study	O
,	O
we	O
aimed	O
to	O
investigate	O
the	O
role	O
of	O
aberrations	O
in	O
the	O
APC	B-Gene_or_gene_product
and	O
beta	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
genes	O
in	O
the	O
pathogenesis	O
of	O
CRC	B-Cancer
in	O
the	O
Kashmir	O
valley	O
,	O
and	O
to	O
correlate	O
it	O
with	O
various	O
clinicopathological	O
variables	O
.	O

We	O
examined	O
the	O
paired	O
tumour	B-Cancer
and	O
normal	O
-	O
tissue	B-Tissue
specimens	I-Tissue
of	O
86	O
CRC	B-Cancer
patients	B-Organism
for	O
the	O
occurrence	O
of	O
aberrations	O
in	O
the	O
mutation	O
cluster	O
region	O
(	O
MCR	O
)	O
of	O
the	O
APC	B-Gene_or_gene_product
gene	O
and	O
exon	O
3	O
of	O
the	O
beta	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
gene	O
by	O
polymerase	O
chain	O
reaction	O
-	O
single	O
-	O
strand	O
conformation	O
polymorphism	O
(	O
PCR	O
-	O
SSCP	O
)	O
and	O
/	O
or	O
PCR	O
-	O
direct	O
sequencing	O
.	O

Analysis	O
of	O
promoter	O
hypermethylation	O
of	O
the	O
APC	B-Gene_or_gene_product
gene	O
was	O
also	O
carried	O
out	O
using	O
methylation	O
-	O
specific	O
PCR	O
(	O
MS	O
-	O
PCR	O
)	O
.	O

The	O
overall	O
mutation	O
rate	O
of	O
the	O
MCR	O
of	O
the	O
APC	B-Gene_or_gene_product
gene	O
among	O
86	O
CRC	B-Cancer
cases	O
was	O
12	O
.	O
8	O
per	O
cent	O
(	O
11	O
of	O
86	O
)	O
.	O

Promoter	O
hypermethylation	O
of	O
APC	B-Gene_or_gene_product
was	O
observed	O
in	O
54	O
.	O
65	O
per	O
cent	O
(	O
47	O
of	O
86	O
)	O
of	O
cases	O
.	O

Furthermore	O
,	O
we	O
found	O
a	O
significant	O
association	O
between	O
tumour	B-Cancer
location	O
,	O
tumour	B-Cancer
grade	O
and	O
node	B-Multi-tissue_structure
status	O
and	O
the	O
methylation	O
status	O
of	O
the	O
APC	B-Gene_or_gene_product
gene	O
(	O
p	O
<	O
=	O
0	O
.	O
05	O
)	O
.	O

Although	O
the	O
number	O
of	O
mutations	O
in	O
the	O
APC	B-Gene_or_gene_product
and	O
beta	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
genes	O
in	O
our	O
CRC	B-Cancer
cases	O
was	O
very	O
low	O
,	O
the	O
study	O
confirms	O
the	O
role	O
of	O
epigenetic	O
gene	O
silencing	O
of	O
the	O
pivotal	O
molecular	O
gladiator	O
,	O
APC	B-Gene_or_gene_product
,	O
of	O
the	O
Wnt	B-Gene_or_gene_product
pathway	O
in	O
the	O
development	O
of	O
CRC	B-Cancer
in	O
the	O
Kashmiri	O
population	O
.	O

Aglycon	B-Simple_chemical
of	O
rhizochalin	B-Simple_chemical
from	O
the	O
Rhizochalina	B-Organism
incrustata	I-Organism
induces	O
apoptosis	O
via	O
activation	O
of	O
AMP	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
in	O
HT	B-Cell
-	I-Cell
29	I-Cell
colon	I-Cell
cancer	I-Cell
cells	I-Cell
.	O

Rhizochalin	B-Simple_chemical
is	O
a	O
two	O
-	O
headed	O
sphingolipid	B-Simple_chemical
-	O
like	O
compound	O
isolated	O
from	O
the	O
sponge	O
Rhizochalina	B-Organism
incrustata	I-Organism
.	O

It	O
has	O
been	O
reported	O
that	O
rhizocalin	B-Simple_chemical
and	O
its	O
derivates	O
have	O
a	O
chemopreventive	O
and	O
chemotherapeutic	O
effect	O
.	O

However	O
,	O
the	O
molecular	O
mechanism	O
of	O
these	O
effects	O
is	O
not	O
understood	O
.	O

Here	O
,	O
we	O
demonstrate	O
that	O
aglycon	B-Simple_chemical
of	O
rhizochalin	B-Simple_chemical
(	O
AglRhz	B-Simple_chemical
)	O
from	O
the	O
Rhizochalina	B-Organism
incrustata	I-Organism
induces	O
AMP	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
AMPK	B-Gene_or_gene_product
)	O
phosphorylation	O
,	O
and	O
thereby	O
inhibits	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
-	O
p70S6	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
-	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
signaling	O
and	O
activator	B-Gene_or_gene_product
protein	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
activity	O
via	O
phosphorylation	O
of	O
Raptor	B-Gene_or_gene_product
in	O
HT	B-Cell
-	I-Cell
29	I-Cell
cells	I-Cell
.	O

In	O
addition	O
,	O
AglRhz	B-Simple_chemical
induced	O
activation	O
of	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
and	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
,	O
and	O
DNA	B-Cellular_component
fragmentation	O
in	O
HT	B-Cell
-	I-Cell
29	I-Cell
cells	I-Cell
,	O
leads	O
to	O
induction	O
of	O
apoptosis	O
as	O
well	O
as	O
suppression	O
of	O
tumorigenicity	O
of	O
HT	B-Cell
-	I-Cell
29	I-Cell
cells	I-Cell
.	O

Notably	O
,	O
AglRhz	B-Simple_chemical
inhibits	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
(	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
-	O
induced	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
activity	O
and	O
cell	B-Cell
transformation	O
in	O
JB6	B-Cell
Cl41	I-Cell
cells	I-Cell
.	O

Overall	O
,	O
our	O
findings	O
identify	O
AMPK	B-Gene_or_gene_product
as	O
an	O
important	O
target	O
protein	O
for	O
mediating	O
the	O
anti	O
-	O
tumor	B-Cancer
properties	O
of	O
AglRhz	B-Simple_chemical
in	O
HT	B-Cell
-	I-Cell
29	I-Cell
colon	I-Cell
cancer	I-Cell
cells	I-Cell
and	O
have	O
important	O
implication	O
for	O
sponges	O
,	O
the	O
most	O
important	O
marine	O
source	O
,	O
in	O
colon	B-Cancer
cancer	I-Cancer
.	O

Expression	O
of	O
thymidylate	B-Gene_or_gene_product
synthase	I-Gene_or_gene_product
and	O
dihydropyrimidine	B-Gene_or_gene_product
dehydrogenase	I-Gene_or_gene_product
in	O
primary	B-Cancer
oral	I-Cancer
squamous	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
and	O
corresponding	O
metastases	O
in	O
cervical	B-Multi-tissue_structure
lymph	I-Multi-tissue_structure
nodes	I-Multi-tissue_structure
:	O
association	O
with	O
the	O
metastasis	O
suppressor	O
CD82	B-Gene_or_gene_product
.	O

Thymidylate	B-Gene_or_gene_product
synthase	I-Gene_or_gene_product
(	O
TS	B-Gene_or_gene_product
)	O
and	O
dihydropyrimidine	B-Gene_or_gene_product
dehydrogenase	I-Gene_or_gene_product
(	O
DPD	B-Gene_or_gene_product
)	O
are	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
metabolizing	O
enzymes	O
and	O
are	O
involved	O
in	O
the	O
sensitivity	O
of	O
carcinoma	B-Cancer
patients	B-Organism
to	O
5	B-Simple_chemical
-	I-Simple_chemical
FU	I-Simple_chemical
.	O

Although	O
5	B-Simple_chemical
-	I-Simple_chemical
FU	I-Simple_chemical
is	O
often	O
used	O
for	O
the	O
treatment	O
of	O
oral	B-Cancer
carcinoma	I-Cancer
,	O
there	O
has	O
not	O
been	O
any	O
investigation	O
into	O
the	O
expression	O
of	O
these	O
enzymes	O
in	O
metastatic	B-Multi-tissue_structure
lymph	I-Multi-tissue_structure
nodes	I-Multi-tissue_structure
or	O
of	O
their	O
roles	O
in	O
the	O
effectiveness	O
of	O
5	B-Simple_chemical
-	I-Simple_chemical
FU	I-Simple_chemical
in	O
treating	O
lymph	B-Cancer
node	I-Cancer
-	I-Cancer
metastatic	I-Cancer
cancer	I-Cancer
.	O

Oral	B-Cancer
squamous	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
(	O
OSCC	B-Cancer
)	O
often	O
metastasizes	O
to	O
the	O
lymph	B-Multi-tissue_structure
nodes	I-Multi-tissue_structure
,	O
and	O
these	O
enzymes	O
may	O
be	O
significant	O
in	O
the	O
survival	O
of	O
patients	B-Organism
with	O
this	O
disease	O
.	O

This	O
study	O
investigated	O
the	O
expression	O
of	O
TS	B-Gene_or_gene_product
and	O
DPD	B-Gene_or_gene_product
in	O
cervical	B-Multi-tissue_structure
lymph	I-Multi-tissue_structure
node	I-Multi-tissue_structure
metastases	O
and	O
its	O
relationship	O
with	O
primary	B-Cancer
OSCC	I-Cancer
,	O
as	O
well	O
as	O
the	O
interaction	O
between	O
these	O
enzymes	O
and	O
Kangai	B-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
KAI1	B-Gene_or_gene_product
/	O
CD82	B-Gene_or_gene_product
)	O
which	O
is	O
a	O
metastasis	O
suppressor	O
protein	O
.	O

Surgical	O
specimens	B-Multi-tissue_structure
from	O
20	O
cases	O
of	O
OSCC	B-Cancer
with	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
,	O
20	O
cases	O
of	O
OSCC	B-Cancer
without	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
,	O
and	O
10	O
cases	O
of	O
normal	O
mucosa	B-Multi-tissue_structure
were	O
examined	O
by	O
immunohistochemistry	O
.	O

The	O
relationship	O
between	O
TS	B-Gene_or_gene_product
and	O
DPD	B-Gene_or_gene_product
expression	O
and	O
clinicopathological	O
data	O
was	O
analyzed	O
.	O

TS	B-Gene_or_gene_product
and	O
DPD	B-Gene_or_gene_product
proteins	O
were	O
overexpressed	O
in	O
primary	B-Cancer
OSCC	I-Cancer
compared	O
to	O
that	O
in	O
normal	O
mucosa	B-Multi-tissue_structure
.	O

TS	B-Gene_or_gene_product
expression	O
of	O
the	O
primary	O
oral	B-Cell
cancer	I-Cell
cells	I-Cell
in	O
the	O
group	O
with	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
was	O
higher	O
than	O
that	O
of	O
those	O
without	O
.	O

DPD	B-Gene_or_gene_product
expression	O
did	O
not	O
significantly	O
correlate	O
with	O
the	O
occurrence	O
of	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
,	O
nor	O
was	O
it	O
different	O
between	O
primary	O
oral	B-Cell
cancer	I-Cell
cells	I-Cell
and	O
cervical	B-Multi-tissue_structure
metastases	O
.	O

CD82	B-Gene_or_gene_product
expression	O
was	O
significantly	O
reduced	O
in	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastases	O
.	O

These	O
findings	O
indicate	O
that	O
TS	B-Gene_or_gene_product
and	O
CD82	B-Gene_or_gene_product
may	O
be	O
of	O
great	O
value	O
in	O
assessing	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
of	O
OSCC	B-Cancer
,	O
and	O
could	O
be	O
taken	O
as	O
new	O
targets	O
for	O
therapy	O
of	O
metastatic	O
OSCC	B-Cancer
.	O

Impaired	O
CK1	B-Gene_or_gene_product
delta	I-Gene_or_gene_product
activity	O
attenuates	O
SV40	B-Organism
-	O
induced	O
cellular	B-Cell
transformation	O
in	O
vitro	O
and	O
mouse	B-Organism
mammary	B-Organ
carcinogenesis	O
in	O
vivo	O
.	O

Simian	B-Organism
virus	I-Organism
40	I-Organism
(	O
SV40	B-Organism
)	O
is	O
a	O
powerful	O
tool	O
to	O
study	O
cellular	B-Cell
transformation	O
in	O
vitro	O
,	O
as	O
well	O
as	O
tumor	B-Cancer
development	O
and	O
progression	O
in	O
vivo	O
.	O

Various	O
cellular	B-Cell
kinases	O
,	O
among	O
them	O
members	O
of	O
the	O
CK1	B-Gene_or_gene_product
family	O
,	O
play	O
an	O
important	O
role	O
in	O
modulating	O
the	O
transforming	O
activity	O
of	O
SV40	B-Organism
,	O
including	O
the	O
transforming	O
activity	O
of	O
T	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Ag	I-Gene_or_gene_product
,	O
the	O
major	O
transforming	O
protein	O
of	O
SV40	B-Organism
,	O
itself	O
.	O

Here	O
we	O
characterized	O
the	O
effects	O
of	O
mutant	O
CK1delta	B-Gene_or_gene_product
variants	O
with	O
impaired	O
kinase	O
activity	O
on	O
SV40	B-Organism
-	O
induced	O
cell	B-Cell
transformation	O
in	O
vitro	O
,	O
and	O
on	O
SV40	B-Organism
-	O
induced	O
mammary	B-Organ
carcinogenesis	O
in	O
vivo	O
in	O
a	O
transgenic	B-Organism
/	I-Organism
bi	I-Organism
-	I-Organism
transgenic	I-Organism
mouse	I-Organism
model	O
.	O

CK1delta	B-Gene_or_gene_product
mutants	O
exhibited	O
a	O
reduced	O
kinase	O
activity	O
compared	O
to	O
wtCK1delta	B-Gene_or_gene_product
in	O
in	O
vitro	O
kinase	O
assays	O
.	O

Molecular	O
modeling	O
studies	O
suggested	O
that	O
mutation	O
N172D	O
,	O
located	O
within	O
the	O
substrate	O
binding	O
region	O
,	O
is	O
mainly	O
responsible	O
for	O
impaired	O
mutCK1delta	B-Gene_or_gene_product
activity	O
.	O

When	O
stably	O
over	O
-	O
expressed	O
in	O
maximal	O
transformed	O
SV	B-Cell
-	I-Cell
52	I-Cell
cells	I-Cell
,	O
CK1delta	B-Gene_or_gene_product
mutants	O
induced	O
reversion	O
to	O
a	O
minimal	O
transformed	O
phenotype	O
by	O
dominant	O
-	O
negative	O
interference	O
with	O
endogenous	O
wtCK1delta	B-Gene_or_gene_product
.	O

To	O
characterize	O
the	O
effects	O
of	O
CK1delta	B-Gene_or_gene_product
on	O
SV40	B-Organism
-	O
induced	O
mammary	B-Organ
carcinogenesis	O
,	O
we	O
generated	O
transgenic	B-Organism
mice	I-Organism
expressing	O
mutant	O
CK1delta	B-Gene_or_gene_product
under	O
the	O
control	O
of	O
the	O
whey	B-Gene_or_gene_product
acidic	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
(	O
WAP	B-Gene_or_gene_product
)	O
gene	O
promoter	O
,	O
and	O
crossed	O
them	O
with	O
SV40	B-Organism
transgenic	O
WAP	B-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
(	O
WAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
T	I-Gene_or_gene_product
)	O
mice	O
.	O

Both	O
WAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
T	I-Gene_or_gene_product
mice	B-Organism
as	O
well	O
as	O
WAP	B-Gene_or_gene_product
-	O
mutCK1delta	B-Gene_or_gene_product
/	O
WAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
T	I-Gene_or_gene_product
bi	O
-	O
transgenic	O
mice	O
developed	O
breast	B-Cancer
cancer	I-Cancer
.	O

However	O
,	O
tumor	B-Cancer
incidence	O
was	O
lower	O
and	O
life	O
span	O
was	O
significantly	O
longer	O
in	O
WAP	B-Gene_or_gene_product
-	O
mutCK1delta	B-Gene_or_gene_product
/	O
WAP	B-Gene_or_gene_product
-	O
T	O
bi	O
-	O
transgenic	O
animals	O
.	O

The	O
reduced	O
CK1delta	B-Gene_or_gene_product
activity	O
did	O
not	O
affect	O
early	O
lesion	B-Pathological_formation
formation	O
during	O
tumorigenesis	O
,	O
suggesting	O
that	O
impaired	O
CK1delta	B-Gene_or_gene_product
activity	O
reduces	O
the	O
probability	O
for	O
outgrowth	O
of	O
in	B-Cancer
situ	I-Cancer
carcinomas	I-Cancer
to	O
invasive	B-Cancer
carcinomas	I-Cancer
.	O

The	O
different	O
tumorigenic	O
potential	O
of	O
SV40	B-Organism
in	O
WAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
T	I-Gene_or_gene_product
and	O
WAP	B-Gene_or_gene_product
-	O
mutCK1delta	B-Gene_or_gene_product
/	O
WAP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
T	I-Gene_or_gene_product
tumors	B-Cancer
was	O
also	O
reflected	O
by	O
a	O
significantly	O
different	O
expression	O
of	O
various	O
genes	O
known	O
to	O
be	O
involved	O
in	O
tumor	B-Cancer
progression	O
,	O
specifically	O
of	O
those	O
involved	O
in	O
wnt	B-Gene_or_gene_product
-	O
signaling	O
and	O
DNA	B-Cellular_component
repair	O
.	O

Our	O
data	O
show	O
that	O
inactivating	O
mutations	O
in	O
CK1delta	B-Gene_or_gene_product
impair	O
SV40	B-Organism
-	O
induced	O
cellular	B-Cell
transformation	O
in	O
vitro	O
and	O
mouse	B-Organism
mammary	B-Organ
carcinogenesis	O
in	O
vivo	O
.	O

Osteonectin	B-Gene_or_gene_product
transcript	O
and	O
metastatic	O
behavior	O
in	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Ki	I-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
transformed	O
fibroblasts	B-Cell
.	O

Osteonectin	B-Gene_or_gene_product
is	O
one	O
of	O
the	O
major	O
non	O
-	O
collagenous	O
proteins	O
of	O
bone	B-Tissue
.	O

However	O
,	O
its	O
transcript	O
has	O
been	O
found	O
in	O
many	O
soft	B-Tissue
,	I-Tissue
extracellular	I-Tissue
matrix	I-Tissue
-	I-Tissue
producing	I-Tissue
tissues	I-Tissue
;	O
an	O
osteonectin	B-Gene_or_gene_product
-	O
related	O
protein	O
was	O
detected	O
in	O
tumor	B-Cellular_component
basement	I-Cellular_component
membrane	I-Cellular_component
.	O

We	O
have	O
investigated	O
the	O
expression	O
of	O
osteonectin	B-Gene_or_gene_product
gene	O
in	O
fresh	O
BALB	B-Cell
/	I-Cell
c	I-Cell
fibroblasts	I-Cell
transformed	O
by	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Ki	I-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
.	O

Transformed	O
cells	B-Cell
exhibited	O
lower	O
levels	O
of	O
RNA	O
as	O
compared	O
with	O
normal	O
fibroblasts	B-Cell
.	O

The	O
transformed	O
cells	B-Cell
were	O
cloned	O
after	O
in	O
vivo	O
tumorigenic	O
assay	O
,	O
and	O
4	O
clones	B-Cell
were	O
analyzed	O
for	O
osteonectin	B-Gene_or_gene_product
expression	O
by	O
Northern	O
blots	O
.	O

Two	O
of	O
them	O
were	O
selected	O
for	O
high	O
or	O
low	O
osteonectin	B-Gene_or_gene_product
expression	O
and	O
tested	O
in	O
vivo	O
in	O
spontaneous	O
and	O
artificial	O
metastasis	O
assays	O
.	O

High	O
osteonectin	B-Gene_or_gene_product
expression	O
was	O
correlated	O
with	O
high	O
lung	B-Organ
colonization	O
.	O

When	O
10	O
(	O
5	O
)	O
cells	B-Cell
were	O
injected	O
i	B-Immaterial_anatomical_entity
.	I-Immaterial_anatomical_entity
v	I-Immaterial_anatomical_entity
.	I-Immaterial_anatomical_entity
,	O
median	O
colony	B-Cell
value	O
was	O
55	O
and	O
20	O
in	O
higher	O
expressor	O
vs	O
.	O
lower	O
expressor	O
respectively	O
(	O
p	O
less	O
than	O
0	O
.	O
005	O
)	O
.	O

Spontaneous	O
metastasis	O
indicates	O
a	O
possible	O
reverse	O
correlation	O
.	O

Our	O
data	O
align	O
osteonectin	B-Gene_or_gene_product
with	O
other	O
matrix	B-Cellular_component
-	O
components	O
and	O
adhesion	O
molecules	O
in	O
affecting	O
potential	O
metastatic	O
spreading	O
of	O
transformed	B-Cell
cells	I-Cell
.	O

[	O
Intraepithelial	B-Cancer
neoplasm	I-Cancer
of	O
the	O
uterine	B-Multi-tissue_structure
cervix	I-Multi-tissue_structure
and	O
angiogenesis	O
:	O
morphologic	O
study	O
]	O

Thirty	O
uterine	B-Multi-tissue_structure
cervix	I-Multi-tissue_structure
specimens	I-Multi-tissue_structure
sampled	O
following	O
conization	O
or	O
total	O
hysterectomy	O
were	O
studied	O
using	O
histology	O
,	O
histoenzymology	O
(	O
vessel	B-Multi-tissue_structure
phosphatase	O
alkaline	O
activity	O
)	O
,	O
and	O
immunohistochemistry	O
(	O
demonstration	O
of	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
in	O
epithelium	B-Tissue
and	O
vessel	B-Cellular_component
basement	I-Cellular_component
membranes	I-Cellular_component
)	O
.	O

Pathologic	O
conditions	O
included	O
dystrophia	O
,	O
moderate	O
dysplasia	O
,	O
severe	O
dysplasia	O
,	O
and	O
intraepithelial	B-Cancer
carcinoma	I-Cancer
.	O

Results	O
were	O
compared	O
to	O
findings	O
in	O
a	O
control	O
group	O
.	O

We	O
found	O
that	O
the	O
severity	O
of	O
vascular	B-Multi-tissue_structure
abnormalities	O
correlated	O
positively	O
with	O
the	O
severity	O
of	O
histologic	O
epithelial	B-Pathological_formation
lesions	I-Pathological_formation
;	O
this	O
finding	O
is	O
consistent	O
with	O
colposcopic	O
results	O
.	O

Anarchic	O
angiogenesis	O
with	O
large	O
,	O
moniliform	O
,	O
tortuous	O
vessels	B-Multi-tissue_structure
was	O
seen	O
in	O
severe	O
dysplasias	O
and	O
carcinomas	B-Cancer
.	O

The	O
vascular	B-Multi-tissue_structure
anomalies	O
seem	O
to	O
precede	O
the	O
development	O
of	O
histologic	B-Pathological_formation
lesions	I-Pathological_formation
in	O
some	O
instances	O
.	O

Histogenesis	O
of	O
the	O
abnormal	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
may	O
involve	O
production	O
of	O
an	O
angiogenic	O
factor	O
by	O
the	O
cancerized	B-Tissue
epithelia	I-Tissue
.	O

Glycolysis	O
and	O
glutaminolysis	O
in	O
perifused	O
Ehrlich	B-Cell
ascites	I-Cell
tumour	I-Cell
cells	I-Cell
.	O

A	O
perifusion	O
system	O
was	O
designed	O
in	O
order	O
to	O
study	O
glucose	B-Simple_chemical
and	O
glutamine	B-Amino_acid
metabolism	O
by	O
freshly	O
harvested	O
Ehrlich	B-Cell
ascites	I-Cell
tumour	I-Cell
cells	I-Cell
in	O
steady	O
state	O
conditions	O
.	O

Cells	B-Cell
were	O
perifused	O
in	O
the	O
presence	O
of	O
5	O
mM	O
glucose	B-Simple_chemical
,	O
0	O
.	O
5	O
mM	O
glutamine	B-Amino_acid
or	O
5	O
mM	O
glucose	B-Simple_chemical
and	O
0	O
.	O
5	O
mM	O
glutamine	B-Amino_acid
.	O

The	O
results	O
in	O
steady	O
state	O
reveal	O
that	O
both	O
substrates	O
glucose	B-Simple_chemical
and	O
glutamine	B-Amino_acid
are	O
continuously	O
wasted	O
by	O
tumour	B-Cell
cells	I-Cell
,	O
excreting	O
two	O
moles	O
of	O
lactate	B-Simple_chemical
per	O
mol	O
of	O
glucose	B-Simple_chemical
and	O
one	O
mol	O
of	O
glutamate	B-Amino_acid
and	O
ammonia	B-Simple_chemical
per	O
mol	O
of	O
glutamine	B-Amino_acid
consumed	O
into	O
the	O
medium	O
.	O

Glutamine	B-Amino_acid
consumption	O
in	O
the	O
presence	O
of	O
glucose	B-Simple_chemical
was	O
higher	O
than	O
with	O
glutamine	B-Amino_acid
alone	O
.	O

Multi	O
-	O
step	O
neoplastic	O
transformation	O
of	O
normal	O
human	B-Organism
fibroblasts	B-Cell
by	O
Co	B-Simple_chemical
-	I-Simple_chemical
60	I-Simple_chemical
gamma	O
rays	O
and	O
Ha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
oncogenes	O
.	O

As	O
reported	O
previously	O
(	O
Namba	O
et	O
al	O
.	O
,	O
1985	O
;	O
Namba	O
,	O
1985	O
)	O
,	O
normal	O
human	B-Organism
fibroblasts	B-Cell
were	O
transformed	O
into	O
immortal	B-Cell
cells	I-Cell
with	O
abnormal	O
karyotypes	O
by	O
Co	B-Simple_chemical
-	I-Simple_chemical
60	I-Simple_chemical
gamma	O
-	O
ray	O
irradiation	O
.	O

These	O
immortally	O
transformed	O
cells	B-Cell
(	O
KMST	B-Cell
-	I-Cell
6	I-Cell
)	O
showed	O
no	O
clonability	O
in	O
soft	O
agar	O
and	O
were	O
not	O
tumorigenic	O
.	O

However	O
,	O
by	O
treatment	O
with	O
Ha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
oncogenes	O
derived	O
from	O
a	O
human	B-Organism
lung	I-Organism
carcinoma	I-Organism
or	O
Harvey	B-Organism
murine	I-Organism
sarcoma	I-Organism
virus	I-Organism
,	O
the	O
KMST	B-Cell
-	I-Cell
6	I-Cell
cells	I-Cell
acquired	O
elevated	O
clonability	O
in	O
soft	O
agar	O
and	O
transplantability	O
in	O
nude	B-Organism
mice	I-Organism
.	O

All	O
the	O
tumors	B-Cancer
produced	O
grew	O
progressively	O
without	O
showing	O
regression	O
and	O
killed	O
the	O
mice	B-Organism
.	O

The	O
tumors	B-Cancer
were	O
also	O
serially	O
transplantable	O
into	O
other	O
mice	B-Organism
.	O

The	O
Ha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
oncogene	O
alone	O
did	O
not	O
convert	O
normal	O
human	B-Organism
fibroblasts	B-Cell
into	O
either	O
immortal	B-Cell
or	O
tumorigenic	B-Cell
cells	I-Cell
.	O

Our	O
current	O
data	O
suggest	O
that	O
gamma	O
rays	O
worked	O
as	O
an	O
initiator	O
of	O
carcinogenesis	O
in	O
normal	O
human	B-Organism
cells	B-Cell
,	O
giving	O
rise	O
to	O
chromosome	B-Cellular_component
aberrations	O
and	O
immortality	O
,	O
and	O
the	O
Ha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
oncogene	O
played	O
a	O
role	O
in	O
the	O
progression	O
of	O
the	O
immortally	O
transformed	O
cell	B-Cell
population	O
to	O
a	O
neoplastic	O
one	O
showing	O
enhanced	O
colony	O
formation	O
in	O
soft	O
agar	O
and	O
tumorigenicity	O
in	O
nude	B-Organism
mice	I-Organism
.	O

New	O
functions	O
of	O
epidermal	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
:	O
stimulation	O
of	O
capillary	B-Cell
endothelial	I-Cell
cell	I-Cell
migration	O
and	O
matrix	B-Cellular_component
dependent	O
proliferation	O
.	O

The	O
proliferative	O
response	O
of	O
bovine	B-Organism
retinal	B-Cell
capillary	I-Cell
endothelial	I-Cell
cells	I-Cell
to	O
EGF	B-Gene_or_gene_product
is	O
dependent	O
upon	O
attaching	O
the	O
cells	B-Cell
to	O
a	O
matrix	B-Cellular_component
of	O
fibronectin	B-Gene_or_gene_product
.	O

Bovine	B-Organism
capillary	B-Cell
endothelial	I-Cell
cells	I-Cell
are	O
also	O
stimulated	O
to	O
actively	O
migrate	O
when	O
exposed	O
to	O
EGF	B-Gene_or_gene_product
in	O
vitro	O
.	O

These	O
activities	O
provide	O
an	O
explanation	O
for	O
the	O
angiogenic	O
properties	O
of	O
EGF	B-Gene_or_gene_product
in	O
vivo	O
.	O

Capillary	B-Cell
cell	I-Cell
migration	O
and	O
proliferation	O
are	O
proposed	O
as	O
sensitive	O
quantifiable	O
bioassays	O
to	O
explore	O
the	O
functional	O
domains	O
of	O
the	O
EGF	B-Gene_or_gene_product
molecule	O
.	O

Studies	O
on	O
the	O
inactivation	O
of	O
these	O
properties	O
of	O
EGF	B-Gene_or_gene_product
by	O
specific	O
cleavage	O
of	O
the	O
molecule	O
with	O
CNBr	B-Simple_chemical
or	O
proteases	O
suggest	O
that	O
an	O
intact	O
loop	O
composed	O
in	O
part	O
by	O
amino	O
acid	O
residues	O
20	O
to	O
31	O
is	O
essential	O
for	O
at	O
least	O
some	O
functions	O
.	O

Angiogenesis	O
:	O
initiation	O
and	O
control	O
.	O

From	O
in	O
vivo	O
experiments	O
using	O
new	O
methods	O
such	O
as	O
the	O
rabbit	B-Organism
cornea	B-Multi-tissue_structure
,	O
it	O
is	O
now	O
becoming	O
clear	O
that	O
the	O
growth	O
of	O
a	O
capillary	B-Tissue
involves	O
an	O
ordered	O
sequence	O
of	O
events	O
that	O
includes	O
lysis	O
of	O
the	O
basement	B-Cellular_component
membrane	I-Cellular_component
of	O
a	O
parent	O
venule	B-Multi-tissue_structure
,	O
directional	O
migration	O
of	O
capillary	B-Cell
endothelial	I-Cell
cells	I-Cell
toward	O
the	O
angiogenic	O
stimulus	O
,	O
lumen	B-Immaterial_anatomical_entity
formation	O
,	O
development	O
of	O
branches	B-Tissue
,	O
and	O
anastomosis	O
of	O
the	O
tip	O
of	O
one	O
tube	B-Tissue
with	O
another	O
to	O
form	O
a	O
loop	O
.	O

It	O
is	O
also	O
clear	O
that	O
diffusible	O
angiogenic	O
stimuli	O
can	O
be	O
released	O
not	O
only	O
from	O
most	O
solid	B-Cancer
tumors	I-Cancer
,	O
but	O
also	O
from	O
at	O
least	O
three	O
non	B-Cell
-	I-Cell
neoplastic	I-Cell
cells	I-Cell
.	O

These	O
include	O
activated	O
macrophages	B-Cell
,	O
sensitized	O
lymphocytes	B-Cell
,	O
and	O
adipocytes	B-Cell
.	O

Other	O
normal	B-Tissue
tissues	I-Tissue
can	O
also	O
stimulate	O
angiogenesis	O
,	O
but	O
the	O
type	O
of	O
cell	B-Cell
giving	O
rise	O
to	O
the	O
angiogenic	O
stimulus	O
is	O
unknown	O
,	O
and	O
the	O
period	O
of	O
angiogenic	O
stimulation	O
is	O
brief	O
.	O

With	O
the	O
recent	O
ability	O
to	O
clone	O
capillary	B-Cell
endothelial	I-Cell
cells	I-Cell
and	O
to	O
carry	O
them	O
in	O
long	O
-	O
term	O
culture	O
,	O
it	O
has	O
been	O
possible	O
to	O
further	O
delineate	O
the	O
mechanism	O
of	O
capillary	B-Tissue
growth	O
.	O

In	O
vitro	O
studies	O
have	O
shown	O
that	O
the	O
mast	B-Cell
cell	I-Cell
seems	O
to	O
behave	O
as	O
a	O
helper	B-Cell
cell	I-Cell
for	O
capillary	B-Cell
endothelial	I-Cell
cells	I-Cell
,	O
in	O
some	O
way	O
speeding	O
up	O
their	O
rate	O
of	O
directional	O
migration	O
.	O

At	O
this	O
writing	O
,	O
heparin	B-Simple_chemical
appears	O
to	O
be	O
the	O
principal	O
mast	B-Cell
cell	I-Cell
factor	O
responsible	O
for	O
this	O
effect	O
on	O
capillary	B-Cell
endothelial	I-Cell
cells	I-Cell
.	O

One	O
theoretical	O
possibility	O
is	O
that	O
mast	B-Cell
cells	I-Cell
may	O
prepare	O
the	O
matrix	B-Cellular_component
,	O
perhaps	O
by	O
slow	O
release	O
of	O
heparin	B-Simple_chemical
,	O
so	O
that	O
capillary	B-Tissue
sprouts	I-Tissue
can	O
more	O
easily	O
move	O
through	O
it	O
toward	O
their	O
angiogenic	O
target	O
.	O

While	O
the	O
study	O
of	O
angiogenesis	O
as	O
a	O
phenomenon	O
is	O
still	O
in	O
an	O
early	O
phase	O
,	O
it	O
has	O
become	O
possible	O
,	O
by	O
using	O
a	O
combination	O
of	O
in	O
vitro	O
and	O
in	O
vivo	O
techniques	O
,	O
to	O
more	O
thoroughly	O
understand	O
the	O
initiation	O
and	O
control	O
of	O
capillary	B-Tissue
growth	O
.	O

Protection	O
from	O
experimental	O
ocular	O
herpetic	O
keratitis	O
by	O
a	O
heat	O
-	O
killed	O
virus	O
vaccine	O
.	O

New	B-Organism
Zealand	I-Organism
white	I-Organism
rabbits	I-Organism
were	O
given	O
limbal	O
inoculations	O
of	O
a	O
heat	O
-	O
killed	O
suspension	O
of	O
herpes	B-Organism
simplex	I-Organism
virus	I-Organism
(	O
HSV	B-Organism
)	O
in	O
a	O
lysate	O
of	O
human	B-Organism
embryonic	B-Cell
kidney	I-Cell
cells	I-Cell
.	O

At	O
intervals	O
of	O
four	O
to	O
14	O
days	O
,	O
the	O
animals	O
were	O
challenged	O
by	O
intrastromal	O
inoculation	O
with	O
10	O
,	O
000	O
plaque	O
-	O
forming	O
units	O
of	O
viable	O
HSV	B-Organism
.	O

Epithelial	B-Tissue
keratitis	O
,	O
disciform	B-Pathological_formation
edema	I-Pathological_formation
,	O
and	O
necrotizing	O
keratitis	O
with	O
neovascularization	O
of	O
the	O
cornea	B-Multi-tissue_structure
developed	O
in	O
control	O
animals	O
.	O

Epithelial	B-Tissue
keratitis	O
and	O
corneal	B-Pathological_formation
edema	I-Pathological_formation
also	O
developed	O
in	O
the	O
immunized	O
animals	O
during	O
the	O
first	O
week	O
after	O
virus	O
challenge	O
,	O
but	O
these	O
symptoms	O
rapidly	O
resolved	O
during	O
the	O
following	O
weeks	O
.	O

The	O
absence	O
of	O
iritis	O
,	O
neovascularization	O
,	O
and	O
necrotizing	O
keratitis	O
in	O
the	O
corneas	B-Multi-tissue_structure
of	O
the	O
immunized	O
animals	O
was	O
particularly	O
striking	O
.	O

Protein	O
kinase	O
activities	O
in	O
immune	O
complexes	O
of	O
simian	B-Organism
virus	I-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
and	O
transformation	O
-	O
associated	O
cellular	B-Cell
p53	B-Gene_or_gene_product
protein	O
.	O

Immune	O
complex	O
kinase	O
assays	O
in	O
the	O
simian	B-Organism
virus	I-Organism
40	I-Organism
system	O
were	O
performed	O
by	O
incubation	O
of	O
immunoprecipitates	O
containing	O
tumor	B-Cancer
antigens	O
with	O
[	B-Simple_chemical
gamma	I-Simple_chemical
-	I-Simple_chemical
32P	I-Simple_chemical
]	I-Simple_chemical
ATP	I-Simple_chemical
,	O
followed	O
by	O
analysis	O
of	O
any	O
phosphoacceptor	O
proteins	O
.	O

These	O
assays	O
yielded	O
mainly	O
the	O
viral	B-Organism
large	B-Gene_or_gene_product
T	I-Gene_or_gene_product
-	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
and	O
,	O
in	O
particular	O
,	O
the	O
associated	O
cellular	B-Cell
p53	B-Gene_or_gene_product
as	O
endogenous	O
substrates	O
.	O

The	O
nature	O
of	O
these	O
substrates	O
was	O
confirmed	O
by	O
proteolysis	O
techniques	O
.	O

Under	O
specific	O
conditions	O
,	O
casein	B-Gene_or_gene_product
could	O
be	O
used	O
as	O
an	O
exogenous	O
substrate	O
as	O
well	O
.	O

The	O
kinase	O
reactions	O
showed	O
preference	O
for	O
ATP	B-Simple_chemical
and	O
MgCl2	B-Simple_chemical
instead	O
of	O
GTP	B-Simple_chemical
or	O
MnCl2	B-Simple_chemical
.	O

Both	O
phosphoserine	B-Amino_acid
and	O
phosphothreonine	B-Amino_acid
,	O
but	O
in	O
no	O
case	O
phosphotyrosine	B-Amino_acid
,	O
were	O
detected	O
after	O
an	O
immune	O
complex	O
kinase	O
reaction	O
.	O

Apparently	O
,	O
several	O
in	O
vivo	O
phosphorylation	O
sites	O
were	O
recognized	O
in	O
vitro	O
in	O
both	O
large	B-Gene_or_gene_product
T	I-Gene_or_gene_product
-	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
and	O
p53	B-Gene_or_gene_product
,	O
but	O
the	O
presence	O
of	O
some	O
artifactual	O
sites	O
could	O
not	O
be	O
completely	O
excluded	O
.	O

Although	O
contaminating	O
kinases	O
were	O
detectable	O
in	O
the	O
immune	O
complexes	O
,	O
at	O
least	O
the	O
p53	B-Gene_or_gene_product
molecules	O
were	O
phosphorylated	O
in	O
vitro	O
in	O
a	O
more	O
specific	O
way	O
.	O

This	O
followed	O
from	O
several	O
characteristics	O
of	O
the	O
immune	O
complex	O
kinase	O
reactions	O
and	O
especially	O
from	O
the	O
strong	O
inhibition	O
of	O
p53	B-Gene_or_gene_product
phosphorylation	O
by	O
two	O
anti	B-Gene_or_gene_product
-	I-Gene_or_gene_product
large	I-Gene_or_gene_product
-	I-Gene_or_gene_product
T	I-Gene_or_gene_product
monoclonal	I-Gene_or_gene_product
antibodies	I-Gene_or_gene_product
.	O

It	O
was	O
shown	O
that	O
large	B-Gene_or_gene_product
T	I-Gene_or_gene_product
-	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
showed	O
associated	O
kinase	O
activity	O
,	O
although	O
none	O
of	O
our	O
results	O
could	O
unambiguously	O
demonstrate	O
an	O
intrinsic	O
kinase	O
activity	O
of	O
this	O
protein	O
.	O

Finally	O
,	O
anti	B-Gene_or_gene_product
-	I-Gene_or_gene_product
p53	I-Gene_or_gene_product
monoclonal	I-Gene_or_gene_product
antibodies	I-Gene_or_gene_product
only	O
slightly	O
affected	O
in	O
vitro	O
phosphorylation	O
reactions	O
,	O
whereas	O
a	O
p53	B-Gene_or_gene_product
molecule	O
from	O
a	O
simian	B-Organism
virus	I-Organism
40	I-Organism
-	O
free	O
,	O
chemically	O
transformed	O
human	B-Organism
cell	B-Cell
line	I-Cell
was	O
not	O
phosphorylated	O
in	O
vitro	O
under	O
any	O
condition	O
tested	O
.	O

Thus	O
,	O
it	O
is	O
highly	O
unlikely	O
that	O
the	O
p53	B-Gene_or_gene_product
molecule	O
per	O
se	O
carries	O
intrinsic	O
or	O
even	O
associated	O
kinase	O
activities	O
.	O

Molecular	O
mediators	O
of	O
interactions	O
with	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
components	I-Cellular_component
in	O
metastasis	O
and	O
angiogenesis	O
.	O

Metastasis	O
and	O
tumor	B-Cancer
angiogenesis	O
are	O
invasive	O
phenomena	O
and	O
share	O
many	O
common	O
properties	O
at	O
the	O
physiological	O
level	O
and	O
some	O
similarities	O
at	O
the	O
molecular	O
level	O
.	O

Each	O
consists	O
of	O
repetitive	O
cycles	O
of	O
interaction	O
with	O
adjacent	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
components	I-Cellular_component
by	O
mediating	O
cellular	B-Cell
adhesion	O
,	O
matrix	B-Cellular_component
dissolution	O
,	O
and	O
cellular	B-Cell
motility	O
to	O
achieve	O
metastasis	O
of	O
cancer	B-Cell
cells	I-Cell
or	O
neovascularization	O
of	O
tumors	B-Cancer
.	O

Molecular	O
factors	O
which	O
implement	O
this	O
triad	O
of	O
events	O
are	O
reviewed	O
,	O
as	O
are	O
several	O
signal	O
transduction	O
components	O
which	O
may	O
regulate	O
them	O
.	O

Some	O
potentially	O
promising	O
prognostic	O
,	O
diagnostic	O
,	O
and	O
therapeutic	O
modalities	O
for	O
tumor	B-Cancer
angiogenesis	O
and	O
metastatic	O
disease	O
are	O
also	O
discussed	O
.	O

Suppressed	O
transformation	O
and	O
induced	O
differentiation	O
of	O
HER	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
/	O
neu	B-Gene_or_gene_product
-	O
overexpressing	O
breast	B-Cell
cancer	I-Cell
cells	I-Cell
by	O
emodin	B-Simple_chemical
.	O

The	O
amplification	O
and	O
overexpression	O
of	O
the	O
HER	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
/	O
neu	B-Gene_or_gene_product
proto	O
-	O
oncogene	O
,	O
which	O
encodes	O
the	O
tyrosine	B-Amino_acid
kinase	O
receptor	O
p185neu	B-Gene_or_gene_product
,	O
have	O
been	O
observed	O
frequently	O
in	O
tumors	B-Cancer
from	O
human	B-Organism
breast	B-Cancer
cancer	I-Cancer
patients	B-Organism
and	O
are	O
correlated	O
with	O
poor	O
prognosis	O
.	O

To	O
explore	O
the	O
potential	O
of	O
chemotherapy	O
directed	O
at	O
the	O
tyrosine	B-Amino_acid
kinase	O
of	O
p185neu	B-Gene_or_gene_product
,	O
we	O
have	O
found	O
that	O
emodin	B-Simple_chemical
(	O
3	B-Simple_chemical
-	I-Simple_chemical
methyl	I-Simple_chemical
-	I-Simple_chemical
1	I-Simple_chemical
,	I-Simple_chemical
6	I-Simple_chemical
,	I-Simple_chemical
8	I-Simple_chemical
-	I-Simple_chemical
trihydroxyanthraquinone	I-Simple_chemical
)	O
,	O
a	O
tyrosine	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
inhibitor	O
,	O
suppresses	O
autophosphorylation	O
and	O
transphosphorylation	O
activities	O
of	O
HER	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
/	O
neu	B-Gene_or_gene_product
tyrosine	B-Amino_acid
kinase	O
,	O
resulting	O
in	O
tyrosine	B-Amino_acid
hypophosphorylation	O
of	O
p185neu	B-Gene_or_gene_product
in	O
HER	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
/	O
neu	B-Gene_or_gene_product
-	O
overexpressing	O
breast	B-Cell
cancer	I-Cell
cells	I-Cell
.	O

Emodin	B-Simple_chemical
,	O
at	O
a	O
40	O
-	O
microM	O
concentration	O
,	O
which	O
repressed	O
tyrosine	B-Amino_acid
kinase	O
of	O
p185neu	B-Gene_or_gene_product
,	O
efficiently	O
inhibited	O
both	O
anchorage	O
-	O
dependent	O
and	O
anchorage	O
-	O
independent	O
growth	O
of	O
HER	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
/	O
neu	B-Gene_or_gene_product
-	O
overexpressing	O
breast	B-Cell
cancer	I-Cell
cells	I-Cell
.	O

However	O
,	O
the	O
inhibition	O
was	O
much	O
less	O
effective	O
for	O
those	O
cells	B-Cell
expressing	O
basal	O
levels	O
of	O
p185neu	B-Gene_or_gene_product
under	O
the	O
same	O
conditions	O
.	O

Emodin	B-Simple_chemical
also	O
induced	O
differentiation	O
of	O
HER	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
/	O
neu	B-Gene_or_gene_product
-	O
overexpressing	O
breast	B-Cell
cancer	I-Cell
cells	I-Cell
by	O
exhibiting	O
a	O
morphological	O
maturation	O
property	O
of	O
large	O
lacy	O
nuclei	B-Cellular_component
surrounded	O
by	O
sizable	O
flat	B-Organism_substance
cytoplasm	I-Organism_substance
and	O
by	O
showing	O
a	O
measurable	O
production	O
of	O
large	O
lipid	B-Simple_chemical
droplets	O
,	O
which	O
is	O
a	O
marker	O
of	O
mature	B-Cell
breast	I-Cell
cells	I-Cell
.	O

Therefore	O
,	O
our	O
results	O
indicate	O
that	O
emodin	B-Simple_chemical
inhibits	O
HER	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
/	O
neu	B-Gene_or_gene_product
tyrosine	B-Amino_acid
kinase	O
activity	O
and	O
preferentially	O
suppresses	O
growth	O
and	O
induces	O
differentiation	O
of	O
HER	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
/	O
neu	B-Gene_or_gene_product
-	O
overexpressing	O
cancer	B-Cell
cells	I-Cell
.	O

These	O
results	O
may	O
have	O
chemotherapeutic	O
implications	O
for	O
using	O
emodin	B-Simple_chemical
to	O
target	O
HER	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
/	O
neu	B-Gene_or_gene_product
-	O
overexpressing	O
cancer	B-Cell
cells	I-Cell
.	O

Metastatic	O
NIH	B-Cell
3T3	I-Cell
x	I-Cell
LTA	I-Cell
cell	I-Cell
hybrids	I-Cell
express	O
72	B-Gene_or_gene_product
kDa	I-Gene_or_gene_product
type	I-Gene_or_gene_product
IV	I-Gene_or_gene_product
collagenase	I-Gene_or_gene_product
.	O

We	O
previously	O
reported	O
that	O
the	O
murine	B-Organism
fibroblast	B-Cell
cell	I-Cell
line	I-Cell
LTA	I-Cell
is	O
tumorigenic	O
but	O
non	O
-	O
metastatic	O
,	O
and	O
is	O
non	O
-	O
responsive	O
to	O
a	O
transfected	O
H	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
oncogene	O
.	O

In	O
contrast	O
,	O
NIH	B-Cell
3T3	I-Cell
cells	I-Cell
are	O
non	O
-	O
tumorigenic	O
but	O
are	O
ras	B-Gene_or_gene_product
-	O
responsive	O
and	O
become	O
metastatic	O
when	O
transfected	O
with	O
ras	B-Gene_or_gene_product
.	O

Somatic	B-Cell
cell	I-Cell
hybrids	I-Cell
between	O
LTA	B-Cell
and	O
NIH	B-Cell
3T3	I-Cell
cells	I-Cell
are	O
tumorigenic	O
,	O
metastatic	O
and	O
ras	B-Gene_or_gene_product
-	O
responsive	O
.	O

Here	O
we	O
examined	O
expression	O
of	O
type	B-Gene_or_gene_product
IV	I-Gene_or_gene_product
collagenases	I-Gene_or_gene_product
in	O
parental	O
LTA	B-Cell
and	O
NIH	B-Cell
3T3	I-Cell
cells	I-Cell
(	O
with	O
and	O
without	O
ras	B-Gene_or_gene_product
)	O
and	O
four	O
metastatic	O
LTA	B-Cell
x	I-Cell
NIH	I-Cell
3T3	I-Cell
hybrids	I-Cell
(	O
also	O
with	O
and	O
without	O
ras	B-Gene_or_gene_product
)	O
.	O

Parental	O
NIH	B-Cell
3T3	I-Cell
-	I-Cell
derived	I-Cell
cells	I-Cell
had	O
both	O
72	B-Gene_or_gene_product
kDa	I-Gene_or_gene_product
and	O
92	B-Gene_or_gene_product
kDa	I-Gene_or_gene_product
gelatinase	I-Gene_or_gene_product
activities	O
,	O
and	O
LTA	B-Cell
-	O
derived	O
cells	B-Cell
had	O
either	O
aberrantly	O
sized	O
approximately	O
90	B-Gene_or_gene_product
kDa	I-Gene_or_gene_product
activity	O
alone	O
or	O
neither	O
enzyme	O
activity	O
.	O

All	O
four	O
metastatic	O
hybrids	B-Cell
expressed	O
60	B-Gene_or_gene_product
-	I-Gene_or_gene_product
72	I-Gene_or_gene_product
kDa	I-Gene_or_gene_product
gelatinase	I-Gene_or_gene_product
activity	O
,	O
while	O
three	O
of	O
them	O
also	O
had	O
92	B-Gene_or_gene_product
kDa	I-Gene_or_gene_product
activity	O
and	O
one	O
had	O
only	O
minimal	O
92	B-Gene_or_gene_product
kDa	I-Gene_or_gene_product
activity	O
.	O

Thus	O
the	O
metastatic	O
phenotype	O
of	O
the	O
hybrids	B-Cell
was	O
associated	O
with	O
expression	O
of	O
72	B-Gene_or_gene_product
kDa	I-Gene_or_gene_product
gelatinase	I-Gene_or_gene_product
.	O

Levels	O
of	O
RNA	O
for	O
tissue	B-Tissue
inhibitors	O
of	O
metalloproteinases	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
were	O
relatively	O
constant	O
,	O
suggesting	O
independent	O
regulation	O
of	O
type	B-Gene_or_gene_product
IV	I-Gene_or_gene_product
collagenases	I-Gene_or_gene_product
and	O
their	O
inhibitors	O
.	O

Southern	O
blotting	O
and	O
probing	O
with	O
PCR	O
-	O
synthesized	O
cDNA	O
fragments	O
of	O
the	O
mouse	B-Organism
72	B-Gene_or_gene_product
kDa	I-Gene_or_gene_product
type	I-Gene_or_gene_product
IV	I-Gene_or_gene_product
collagenase	I-Gene_or_gene_product
gene	O
showed	O
that	O
this	O
gene	O
was	O
present	O
in	O
all	O
cells	B-Cell
although	O
the	O
structure	O
of	O
this	O
gene	O
in	O
one	O
of	O
the	O
three	O
LTA	B-Cell
cell	I-Cell
lines	I-Cell
differed	O
from	O
that	O
of	O
the	O
other	O
cells	B-Cell
.	O

Our	O
results	O
suggest	O
that	O
a	O
key	O
change	O
in	O
the	O
metastatic	O
hybrids	B-Cell
,	O
relative	O
to	O
non	O
-	O
metastatic	O
parental	O
LTA	B-Cell
cells	I-Cell
,	O
is	O
induction	O
of	O
expression	O
of	O
72	B-Gene_or_gene_product
kDa	I-Gene_or_gene_product
type	I-Gene_or_gene_product
IV	I-Gene_or_gene_product
collagenase	I-Gene_or_gene_product
.	O

Neoplastic	O
progression	O
of	O
human	B-Organism
colorectal	B-Cancer
cancer	I-Cancer
is	O
associated	O
with	O
overexpression	O
of	O
the	O
stromelysin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
and	O
BM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
40	I-Gene_or_gene_product
/	O
SPARC	B-Gene_or_gene_product
genes	O
.	O

The	O
interaction	O
of	O
neoplastic	B-Cell
cells	I-Cell
with	O
the	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
is	O
a	O
critical	O
event	O
for	O
the	O
initiation	O
of	O
cancer	B-Cancer
invasion	O
and	O
metastasis	O
.	O

This	O
study	O
was	O
designed	O
to	O
evaluate	O
the	O
potential	O
implication	O
of	O
stromelysin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
(	O
ST3	B-Gene_or_gene_product
)	O
,	O
a	O
newly	O
identified	O
member	O
of	O
the	O
matrix	O
-	O
degrading	O
metalloproteinase	O
family	O
,	O
and	O
of	O
BM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
40	I-Gene_or_gene_product
/	O
SPARC	B-Gene_or_gene_product
,	O
a	O
glycoprotein	O
associated	O
with	O
the	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
,	O
during	O
the	O
progression	O
of	O
human	B-Organism
colorectal	B-Cancer
cancers	I-Cancer
.	O

We	O
analyzed	O
the	O
relative	O
abundance	O
of	O
ST3	B-Gene_or_gene_product
and	O
BM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
40	I-Gene_or_gene_product
/	O
SPARC	B-Gene_or_gene_product
transcripts	O
by	O
Northern	O
blot	O
,	O
and	O
their	O
distribution	O
by	O
in	O
situ	O
hybridization	O
,	O
in	O
normal	O
mucosa	B-Multi-tissue_structure
,	O
benign	B-Pathological_formation
adenomas	I-Pathological_formation
,	O
and	O
primary	O
colorectal	B-Cancer
adenocarcinomas	I-Cancer
and	O
their	O
liver	B-Organ
metastases	O
.	O

The	O
ST3	B-Gene_or_gene_product
and	O
BM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
40	I-Gene_or_gene_product
/	O
SPARC	B-Gene_or_gene_product
transcripts	O
were	O
overexpressed	O
in	O
primary	O
colorectal	B-Cancer
cancers	I-Cancer
and	O
their	O
liver	B-Organ
metastases	O
compared	O
to	O
non	O
-	O
neoplastic	O
mucosa	B-Multi-tissue_structure
.	O

These	O
transcripts	O
were	O
localized	O
in	O
stromal	B-Cell
fibroblasts	I-Cell
adjacent	O
to	O
the	O
neoplastic	O
foci	O
.	O

Overexpression	O
of	O
ST3	B-Gene_or_gene_product
correlated	O
with	O
the	O
progression	O
of	O
human	B-Organism
colorectal	B-Cancer
tumors	I-Cancer
toward	O
local	O
invasion	O
and	O
liver	B-Organ
metastasis	O
.	O

Induction	O
of	O
these	O
genes	O
also	O
occurred	O
in	O
diverticulitis	O
and	O
digestive	O
neoplasms	B-Cancer
such	O
as	O
gastric	B-Cancer
and	O
esophageal	B-Cancer
carcinomas	I-Cancer
.	O

Down	O
-	O
regulation	O
of	O
collagen	B-Gene_or_gene_product
XII	I-Gene_or_gene_product
in	O
transformed	O
mesenchymal	B-Cell
cells	I-Cell
.	O

Collagen	B-Gene_or_gene_product
XII	I-Gene_or_gene_product
is	O
a	O
complex	O
multidomain	O
protein	O
associated	O
with	O
the	O
surface	O
of	O
interstitial	B-Organ
collagen	B-Gene_or_gene_product
fibrils	B-Cellular_component
.	O

This	O
protein	O
is	O
produced	O
in	O
large	O
amounts	O
by	O
fibroblasts	B-Cell
cultivated	O
in	O
vitro	O
.	O

However	O
,	O
it	O
is	O
completely	O
absent	O
from	O
cells	B-Cell
transformed	O
by	O
the	O
oncogene	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
or	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
src	I-Gene_or_gene_product
and	O
from	O
cells	B-Cell
derived	O
from	O
a	O
methylcholanthrene	B-Simple_chemical
-	O
induced	O
fibrosarcoma	B-Cancer
.	O

Since	O
all	O
these	O
cells	B-Cell
lack	O
any	O
mRNA	O
for	O
collagen	B-Gene_or_gene_product
,	I-Gene_or_gene_product
XII	I-Gene_or_gene_product
,	O
it	O
seems	O
likely	O
that	O
the	O
synthesis	O
is	O
blocked	O
at	O
the	O
transcriptional	O
level	O
.	O

Experiments	O
with	O
a	O
temperature	O
-	O
sensitive	O
mutant	O
of	O
Rous	B-Organism
sarcoma	I-Organism
virus	I-Organism
demonstrated	O
that	O
a	O
single	O
oncogene	O
product	O
is	O
sufficient	O
to	O
inhibit	O
the	O
synthesis	O
.	O

A	O
reduction	O
in	O
the	O
expression	O
of	O
collagen	B-Gene_or_gene_product
XII	I-Gene_or_gene_product
might	O
have	O
profound	O
effects	O
on	O
the	O
stability	O
of	O
the	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
of	O
transformed	O
cells	B-Cell
.	O

Comparative	O
NMR	O
study	O
of	O
a	O
differentiated	O
rat	B-Organism
hepatoma	B-Cancer
and	O
its	O
dedifferentiated	O
subclone	B-Cell
cultured	O
as	O
spheroids	B-Cell
and	O
as	O
implanted	O
tumors	B-Cancer
.	O

H4IIEC3	B-Cell
(	O
H4	B-Cell
)	O
,	O
a	O
differentiated	O
rat	B-Organism
hepatoma	B-Cell
line	I-Cell
and	O
H5	B-Cell
,	O
its	O
dedifferentiated	O
subclone	B-Cell
,	O
were	O
investigated	O
as	O
proliferating	O
spheroids	B-Cell
and	O
as	O
implanted	O
subcutaneous	B-Cancer
tumors	I-Cancer
in	O
juvenile	B-Organism
rats	I-Organism
.	O

H4	B-Cell
cells	I-Cell
formed	O
tight	O
,	O
round	O
spheroids	B-Cell
whereas	O
H5	B-Cell
cells	I-Cell
formed	O
loose	O
,	O
grape	B-Cell
-	I-Cell
like	I-Cell
structures	I-Cell
.	O

31P	B-Simple_chemical
MR	O
spectra	O
showed	O
that	O
phosphocreatine	B-Simple_chemical
was	O
present	O
in	O
H5	B-Cell
spheroids	I-Cell
but	O
not	O
in	O
H4	B-Cell
spheroids	I-Cell
or	O
tumors	B-Cancer
.	O

[	B-Simple_chemical
13C	I-Simple_chemical
]	I-Simple_chemical
Lactate	I-Simple_chemical
production	O
from	O
[	B-Simple_chemical
13C	I-Simple_chemical
]	I-Simple_chemical
glucose	I-Simple_chemical
,	O
with	O
no	O
detectable	O
uptake	O
of	O
[	B-Simple_chemical
13C	I-Simple_chemical
]	I-Simple_chemical
alanine	I-Simple_chemical
,	O
indicated	O
that	O
energy	O
production	O
in	O
H5	B-Cell
spheroids	I-Cell
was	O
primarily	O
via	O
glycolysis	O
.	O

No	O
[	B-Simple_chemical
13C	I-Simple_chemical
]	I-Simple_chemical
glucose	I-Simple_chemical
utilization	O
was	O
detected	O
in	O
H4	B-Cell
spheroids	I-Cell
,	O
but	O
uptake	O
of	O
alanine	B-Amino_acid
and	O
accumulation	O
of	O
labeled	O
lactate	B-Simple_chemical
,	O
glutamate	B-Amino_acid
and	O
glutamine	B-Amino_acid
indicated	O
oxidation	O
via	O
the	O
tricarboxylic	B-Simple_chemical
acid	I-Simple_chemical
(	O
TCA	B-Simple_chemical
)	O
cycle	O
.	O

Tumors	B-Cancer
of	O
H4	B-Cell
cells	I-Cell
were	O
well	O
perfused	O
,	O
unlike	O
tumors	B-Cancer
of	O
H5	B-Cell
cells	I-Cell
which	O
were	O
highly	O
necrotic	O
.	O

Following	O
i	O
.	O
v	O
.	O
infusion	O
with	O
[	B-Simple_chemical
13C	I-Simple_chemical
]	I-Simple_chemical
alanine	I-Simple_chemical
,	O
[	B-Simple_chemical
13C	I-Simple_chemical
]	I-Simple_chemical
lactate	I-Simple_chemical
and	O
glutamate	B-Amino_acid
,	O
evidence	O
of	O
oxidation	O
via	O
the	O
TCA	B-Simple_chemical
cycle	O
,	O
were	O
observed	O
in	O
H4	B-Cancer
tumors	I-Cancer
.	O

Thus	O
the	O
results	O
obtained	O
by	O
31P	B-Simple_chemical
and	O
13C	B-Simple_chemical
MRS	O
correlated	O
with	O
the	O
differentiation	O
state	O
of	O
H4	B-Cell
and	O
H5	B-Cell
spheroids	I-Cell
and	O
tumors	B-Cancer
.	O

[	O
The	O
erbB	B-Gene_or_gene_product
gene	O
family	O
:	O
significance	O
for	O
tumor	B-Cancer
development	O
,	O
prognosis	O
and	O
new	O
therapeutic	O
modalities	O
]	O
.	O

RNA	B-Organism
and	O
DNA	B-Organism
viruses	I-Organism
can	O
be	O
transforming	O
and	O
tumourigenic	O
agents	O
.	O

The	O
transformation	O
is	O
a	O
consequence	O
of	O
the	O
ability	O
of	O
viruses	B-Organism
to	O
integrate	O
into	O
the	O
host	B-Cell
cell	I-Cell
'	O
s	O
DNA	B-Cellular_component
and	O
to	O
produce	O
transforming	O
proteins	O
.	O

These	O
proteins	O
are	O
mainly	O
produced	O
by	O
specific	O
integral	O
parts	O
of	O
the	O
viral	B-Organism
genome	O
,	O
the	O
oncogenes	O
.	O

Comparison	O
between	O
RNA	O
/	O
DNA	B-Cellular_component
sequence	O
of	O
viral	B-Organism
oncogenes	O
and	O
normal	O
human	B-Organism
genome	O
of	O
non	O
-	O
transformed	O
cells	B-Cell
revealed	O
high	O
sequence	O
similarities	O
in	O
specific	O
genomic	O
areas	O
,	O
which	O
were	O
named	O
cellular	B-Cell
proto	O
-	O
oncogens	O
.	O

They	O
are	O
important	O
components	O
of	O
the	O
growth	O
regulatory	O
pathways	O
in	O
normal	O
cells	B-Cell
.	O

The	O
accumulation	O
of	O
genetic	O
alterations	O
of	O
some	O
proto	O
-	O
oncogens	O
,	O
like	O
the	O
erbB	B-Gene_or_gene_product
-	O
family	O
,	O
may	O
be	O
part	O
of	O
the	O
mechanism	O
,	O
by	O
which	O
malignant	B-Cell
cells	I-Cell
can	O
acquire	O
a	O
selective	O
growth	O
advantage	O
.	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
(	O
EGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
R	I-Gene_or_gene_product
,	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
erbB1	I-Gene_or_gene_product
)	O
,	O
Her	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
/	O
neu	B-Gene_or_gene_product
(	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
erbB2	I-Gene_or_gene_product
)	O
,	O
and	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
erbB3	I-Gene_or_gene_product
are	O
members	O
of	O
the	O
erbB	B-Gene_or_gene_product
-	O
family	O
.	O

The	O
detection	O
of	O
increased	O
abundance	O
of	O
EGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
R	I-Gene_or_gene_product
or	O
Her	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
/	O
neu	B-Gene_or_gene_product
proteins	O
in	O
human	B-Organism
tumours	B-Cancer
can	O
provide	O
additional	O
information	O
on	O
the	O
disease	O
-	O
free	O
survival	O
and	O
overall	O
survival	O
for	O
patients	B-Organism
with	O
breast	B-Cancer
,	O
ovarian	B-Cancer
,	O
endometrial	B-Cancer
or	O
cervical	B-Cancer
cancer	I-Cancer
.	O

Molecular	O
and	O
cell	B-Cell
-	O
physiological	O
analyses	O
have	O
improved	O
the	O
understanding	O
of	O
tumour	B-Cancer
biology	O
and	O
provide	O
the	O
opportunity	O
for	O
new	O
therapeutic	O
approaches	O
.	O

Monoclonal	O
antibody	O
targeted	O
therapy	O
directed	O
against	O
EGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
R	I-Gene_or_gene_product
or	O
Her	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
/	O
neu	B-Gene_or_gene_product
,	O
the	O
use	O
of	O
anti	O
-	O
sense	O
oligonucleotides	O
and	O
oligodeoxynucleotides	O
,	O
and	O
the	O
application	O
of	O
tyrosine	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
and	O
protein	B-Gene_or_gene_product
C	I-Gene_or_gene_product
-	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
inhibitors	O
are	O
currently	O
being	O
investigated	O
.	O

Human	B-Organism
cytomegalovirus	I-Organism
elevates	O
levels	O
of	O
the	O
cellular	B-Cell
protein	O
p53	B-Gene_or_gene_product
in	O
infected	O
fibroblasts	B-Cell
.	O

Human	B-Organism
cytomegalovirus	I-Organism
(	O
HCMV	B-Organism
)	O
,	O
like	O
other	O
DNA	B-Cellular_component
tumor	B-Cancer
viruses	O
,	O
induces	O
morphological	O
transformation	O
of	O
cells	B-Cell
in	O
vitro	O
and	O
stimulates	O
host	O
cell	B-Cell
macromolecular	O
synthesis	O
in	O
infected	O
cells	B-Cell
.	O

Since	O
other	O
DNA	B-Cellular_component
tumor	B-Cancer
viruses	O
,	O
such	O
as	O
simian	B-Organism
virus	I-Organism
40	I-Organism
and	O
adenovirus	B-Organism
,	O
have	O
previously	O
been	O
shown	O
to	O
interact	O
with	O
cellular	B-Cell
protein	O
p53	B-Gene_or_gene_product
,	O
we	O
investigated	O
whether	O
infection	O
of	O
cells	B-Cell
by	O
HCMV	B-Organism
would	O
modulate	O
cellular	B-Cell
p53	B-Gene_or_gene_product
levels	O
.	O

Our	O
results	O
indicate	O
that	O
HCMV	B-Organism
elevates	O
cellular	B-Cell
p53	B-Gene_or_gene_product
levels	O
on	O
the	O
order	O
of	O
10	O
-	O
to	O
20	O
-	O
fold	O
in	O
infected	O
fibroblasts	B-Cell
.	O

The	O
induction	O
of	O
elevated	O
p53	B-Gene_or_gene_product
levels	O
was	O
dependent	O
upon	O
the	O
presence	O
of	O
active	O
virus	O
and	O
was	O
prevented	O
by	O
neutralizing	O
antibody	O
.	O

The	O
induction	O
of	O
elevated	O
p53	B-Gene_or_gene_product
levels	O
was	O
determined	O
not	O
to	O
be	O
due	O
to	O
virus	O
-	O
receptor	O
interactions	O
or	O
HCMV	B-Organism
late	O
events	O
.	O

The	O
induction	O
of	O
elevated	O
p53	B-Gene_or_gene_product
levels	O
commenced	O
at	O
immediate	O
-	O
early	O
times	O
of	O
the	O
HCMV	B-Organism
multiplication	O
cycle	O
(	O
6	O
h	O
postinfection	O
)	O
and	O
reached	O
maximal	O
levels	O
by	O
24	O
h	O
postinfection	O
,	O
before	O
most	O
of	O
the	O
HCMV	B-Organism
DNA	B-Cellular_component
synthesis	O
was	O
initiated	O
.	O

HCMV	B-Organism
immediate	B-Gene_or_gene_product
-	I-Gene_or_gene_product
early	I-Gene_or_gene_product
proteins	I-Gene_or_gene_product
were	O
clearly	O
shown	O
to	O
be	O
responsible	O
for	O
elevating	O
p53	B-Gene_or_gene_product
levels	O
in	O
infected	O
fibroblasts	B-Cell
;	O
expression	O
of	O
HCMV	B-Organism
immediate	B-Gene_or_gene_product
-	I-Gene_or_gene_product
early	I-Gene_or_gene_product
region	I-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
2	B-Gene_or_gene_product
proteins	O
resulted	O
in	O
elevation	O
of	O
p53	B-Gene_or_gene_product
levels	O
in	O
transfected	O
human	B-Organism
fibroblasts	B-Cell
.	O

This	O
is	O
the	O
first	O
report	O
of	O
increased	O
p53	B-Gene_or_gene_product
levels	O
caused	O
by	O
HCMV	B-Organism
in	O
infected	O
fibroblasts	B-Cell
.	O

H	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
mutations	O
in	O
human	B-Organism
pituitary	B-Cancer
carcinoma	I-Cancer
metastases	O
.	O

Molecular	O
mechanisms	O
of	O
pituitary	B-Organ
tumorigenesis	O
were	O
studied	O
using	O
Polymerase	O
chain	O
reaction	O
-	O
single	O
stranded	O
conformational	O
polymorphism	O
with	O
DNA	O
sequencing	O
to	O
identify	O
potential	O
mutations	O
in	O
the	O
ras	B-Gene_or_gene_product
protooncogenes	O
and	O
the	O
tumor	B-Cancer
suppressor	O
gene	O
p53	B-Gene_or_gene_product
in	O
invasive	B-Cancer
pituitary	I-Cancer
adenomas	I-Cancer
and	O
carcinomas	B-Cancer
.	O

Sequencing	O
of	O
exons	O
5	O
through	O
8	O
of	O
the	O
p53	B-Gene_or_gene_product
gene	O
revealed	O
no	O
mutations	O
,	O
nor	O
were	O
mutations	O
detected	O
in	O
the	O
N	B-Gene_or_gene_product
-	I-Gene_or_gene_product
or	O
K	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
protooncogenes	O
in	O
four	O
of	O
the	O
carcinomas	B-Cancer
and	O
their	O
respective	O
metastatic	O
deposits	B-Cancer
.	O

Point	O
mutations	O
of	O
H	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
however	O
,	O
were	O
identified	O
in	O
three	O
distant	O
metastatic	O
pituitary	B-Cancer
tumor	I-Cancer
secondaries	I-Cancer
,	O
but	O
not	O
in	O
their	O
respective	O
primary	B-Cancer
pituitary	I-Cancer
carcinomas	I-Cancer
,	O
or	O
in	O
six	O
invasive	O
adenomas	B-Cancer
.	O

Two	O
of	O
the	O
mutations	O
included	O
a	O
G	B-Simple_chemical
to	O
C	B-Simple_chemical
substitution	O
at	O
codon	O
12	O
,	O
and	O
a	O
G	B-Simple_chemical
to	O
A	B-Simple_chemical
substitution	O
at	O
codon	O
18	O
,	O
resulting	O
in	O
a	O
glycine	B-Amino_acid
to	O
arginine	B-Amino_acid
,	O
and	O
an	O
alanine	B-Amino_acid
to	O
threonine	B-Amino_acid
change	O
at	O
these	O
amino	B-Amino_acid
acids	I-Amino_acid
,	O
respectively	O
.	O

A	O
third	O
mutation	O
involved	O
a	O
single	O
base	O
pair	O
(	O
adenine	B-Simple_chemical
)	O
deletion	O
in	O
codon	O
3	O
of	O
H	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
which	O
causes	O
a	O
frame	O
shift	O
,	O
resulting	O
in	O
a	O
termination	O
signal	O
at	O
codon	O
19	O
.	O

These	O
results	O
suggest	O
that	O
point	O
mutations	O
in	O
p53	B-Gene_or_gene_product
and	O
ras	B-Gene_or_gene_product
are	O
not	O
associated	O
with	O
pituitary	B-Organ
tumorigenesis	O
,	O
however	O
,	O
point	O
mutations	O
of	O
the	O
H	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
gene	O
may	O
be	O
important	O
in	O
the	O
formation	O
and	O
or	O
growth	O
of	O
pituitary	B-Cancer
metastases	I-Cancer
.	O

This	O
observed	O
genomic	O
instability	O
will	O
be	O
of	O
value	O
in	O
predicting	O
the	O
potential	O
metastatic	O
behavior	O
of	O
these	O
aggressive	O
pituitary	B-Cancer
tumors	I-Cancer
.	O

Overexpression	O
of	O
Glut	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
glucose	B-Simple_chemical
transporter	O
in	O
human	B-Organism
breast	B-Cancer
cancer	I-Cancer
.	O

An	O
immunohistochemical	O
study	O
.	O

BACKGROUND	O
:	O
Breast	B-Cancer
cancers	I-Cancer
have	O
higher	O
than	O
normal	O
glucose	B-Simple_chemical
metabolism	O
,	O
but	O
the	O
mechanism	O
of	O
glucose	B-Simple_chemical
entry	O
into	O
these	O
tumors	B-Cancer
is	O
not	O
well	O
understood	O
.	O

METHODS	O
:	O
The	O
expression	O
of	O
five	O
facilitative	O
glucose	B-Simple_chemical
transporters	O
,	O
Glut	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
erythrocyte	B-Cell
type	O
)	O
,	O
Glut	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
(	O
liver	B-Organ
type	O
)	O
,	O
Glut	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
(	O
brain	B-Organ
type	O
)	O
,	O
Glut	B-Gene_or_gene_product
-	I-Gene_or_gene_product
4	I-Gene_or_gene_product
(	O
muscle	B-Organ
/	O
fat	B-Tissue
type	O
)	O
,	O
and	O
Glut	B-Gene_or_gene_product
-	I-Gene_or_gene_product
5	I-Gene_or_gene_product
(	O
small	B-Organ
intestine	I-Organ
type	O
)	O
,	O
was	O
studied	O
by	O
immunohistochemistry	O
of	O
paraffin	B-Simple_chemical
sections	B-Tissue
from	O
12	O
primary	O
human	B-Organism
breast	B-Cancer
cancers	I-Cancer
and	O
8	O
lymph	B-Cancer
node	I-Cancer
metastases	I-Cancer
from	O
2	O
patients	B-Organism
.	O

Rat	B-Organism
tissues	B-Tissue
known	O
to	O
express	O
these	O
glucose	B-Simple_chemical
transporters	O
were	O
used	O
as	O
controls	O
.	O

RESULTS	O
:	O
All	O
the	O
primary	O
breast	B-Cancer
cancers	I-Cancer
and	O
the	O
lymph	B-Cancer
node	I-Cancer
metastases	I-Cancer
were	O
positive	O
for	O
Glut	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

This	O
transporter	O
was	O
expressed	O
on	O
the	O
cell	B-Cellular_component
membrane	I-Cellular_component
and	O
in	O
the	O
cytoplasm	B-Organism_substance
of	O
the	O
tumor	B-Cell
cells	I-Cell
,	O
but	O
exhibited	O
marked	O
intratumoral	B-Cancer
and	O
intertumoral	B-Cancer
variability	O
in	O
the	O
proportions	O
of	O
positive	O
cells	B-Cell
and	O
the	O
intensity	O
of	O
staining	O
.	O

Staining	O
of	O
the	O
normal	O
mammary	B-Tissue
epithelium	I-Tissue
,	O
if	O
present	O
,	O
was	O
much	O
lower	O
than	O
observed	O
in	O
tumor	B-Cell
cells	I-Cell
from	O
the	O
same	O
patient	B-Organism
.	O

Glut	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
was	O
expressed	O
in	O
all	O
of	O
the	O
tumors	B-Cancer
,	O
but	O
the	O
intensity	O
of	O
staining	O
was	O
not	O
consistently	O
stronger	O
than	O
that	O
seen	O
in	O
healthy	B-Tissue
breast	I-Tissue
.	O

Clusters	O
of	O
Glut	B-Gene_or_gene_product
-	I-Gene_or_gene_product
4	I-Gene_or_gene_product
-	O
positive	O
granule	O
were	O
observed	O
in	O
cells	B-Cell
in	O
six	O
of	O
the	O
tumors	B-Cancer
.	O

None	O
of	O
the	O
tumors	B-Cancer
or	O
the	O
healthy	B-Tissue
breast	I-Tissue
in	O
the	O
tissues	B-Tissue
studied	O
expressed	O
Glut	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
or	O
Glut	B-Gene_or_gene_product
-	I-Gene_or_gene_product
5	I-Gene_or_gene_product
.	O

CONCLUSIONS	O
:	O
Higher	O
expression	O
of	O
the	O
glucose	B-Simple_chemical
transporter	O
Glut	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
by	O
breast	B-Cell
cancer	I-Cell
cells	I-Cell
compared	O
with	O
the	O
healthy	B-Tissue
breast	I-Tissue
tissue	I-Tissue
is	O
common	O
.	O

Increased	O
glucose	B-Simple_chemical
transporter	O
protein	O
expression	O
may	O
contribute	O
to	O
the	O
increased	O
uptake	O
of	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
FDG	B-Simple_chemical
)	O
by	O
these	O
tumors	B-Cancer
observed	O
by	O
positron	O
emission	O
tomography	O
(	O
PET	O
)	O
imaging	O
.	O

High	O
density	O
of	O
somatostatin	B-Gene_or_gene_product
receptors	I-Gene_or_gene_product
in	O
veins	B-Multi-tissue_structure
surrounding	O
human	B-Organism
cancer	B-Tissue
tissue	I-Tissue
:	O
role	O
in	O
tumor	B-Cancer
-	O
host	O
interaction	O
?	O

Somatostatin	B-Gene_or_gene_product
receptors	I-Gene_or_gene_product
were	O
detected	O
in	O
peritumoral	B-Multi-tissue_structure
veins	I-Multi-tissue_structure
of	O
various	O
human	B-Organism
cancer	B-Tissue
tissue	I-Tissue
specimens	I-Tissue
.	O

Vascular	B-Tissue
and	O
neoplastic	B-Tissue
tissue	I-Tissue
from	O
14	O
colonic	B-Cancer
adenocarcinomas	I-Cancer
,	O
13	O
carcinoids	B-Cancer
,	O
6	O
renal	B-Cancer
-	I-Cancer
cell	I-Cancer
carcinomas	I-Cancer
and	O
7	O
malignant	B-Cancer
lymphomas	I-Cancer
were	O
analyzed	O
for	O
somatostatin	B-Gene_or_gene_product
receptors	I-Gene_or_gene_product
by	O
use	O
of	O
quantitative	O
receptor	O
autoradiography	O
.	O

In	O
colonic	B-Cancer
carcinoma	I-Cancer
specimens	I-Cancer
,	O
the	O
peritumoral	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
expressed	O
a	O
high	O
density	O
of	O
somatostatin	B-Gene_or_gene_product
receptors	I-Gene_or_gene_product
,	O
whereas	O
the	O
neoplastic	B-Tissue
tissue	I-Tissue
itself	O
was	O
receptor	O
-	O
negative	O
in	O
many	O
cases	O
.	O

In	O
contrast	O
,	O
the	O
incidence	O
and	O
density	O
of	O
somatostatin	B-Gene_or_gene_product
receptors	I-Gene_or_gene_product
in	O
peritumoral	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
was	O
low	O
in	O
well	O
-	O
differentiated	O
gastrointestinal	B-Cancer
and	O
bronchial	B-Cancer
carcinoids	I-Cancer
,	O
in	O
contrast	O
to	O
the	O
high	O
density	O
of	O
such	O
receptors	O
in	O
the	O
carcinoid	B-Tissue
tumor	I-Tissue
tissue	I-Tissue
.	O

Autochthonous	O
vessels	B-Multi-tissue_structure
surrounding	O
other	O
tumors	B-Cancer
such	O
as	O
renal	B-Cancer
-	I-Cancer
cell	I-Cancer
carcinomas	I-Cancer
or	O
malignant	B-Cancer
lymphomas	I-Cancer
also	O
frequently	O
expressed	O
somatostatin	B-Gene_or_gene_product
receptors	I-Gene_or_gene_product
.	O

In	O
all	O
cases	O
,	O
the	O
somatostatin	B-Gene_or_gene_product
receptors	I-Gene_or_gene_product
were	O
localized	O
in	O
veins	B-Multi-tissue_structure
,	O
particularly	O
in	O
the	O
smooth	B-Tissue
-	I-Tissue
muscle	I-Tissue
cell	I-Tissue
layer	I-Tissue
.	O

They	O
exhibited	O
specific	O
and	O
high	O
-	O
affinity	O
binding	O
of	O
somatostatin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
14	I-Gene_or_gene_product
,	O
somatostatin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
28	I-Gene_or_gene_product
and	O
octreotide	B-Simple_chemical
,	O
suggesting	O
a	O
preferential	O
expression	O
of	O
the	O
SSTR2	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
subtype	O
.	O

Since	O
the	O
vessels	B-Multi-tissue_structure
of	O
normal	O
non	O
-	O
neoplastic	O
human	B-Organism
tissues	B-Tissue
,	O
e	O
.	O
g	O
.	O
of	O
intestine	B-Organ
or	O
lymphatic	B-Organ
organs	I-Organ
,	O
have	O
few	O
somatostatin	B-Gene_or_gene_product
receptors	I-Gene_or_gene_product
,	O
the	O
increased	O
somatostatin	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
expression	O
in	O
peritumoral	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
observed	O
in	O
this	O
study	O
may	O
be	O
linked	O
to	O
the	O
neoplastic	O
process	O
itself	O
.	O

The	O
results	O
suggest	O
that	O
somatostatin	B-Gene_or_gene_product
and	O
somatostatin	B-Gene_or_gene_product
receptors	I-Gene_or_gene_product
may	O
play	O
a	O
regulatory	O
role	O
for	O
hemodynamic	O
tumor	B-Cancer
-	O
host	O
interactions	O
,	O
possibly	O
involving	O
tumor	B-Tissue
stroma	I-Tissue
generation	O
,	O
tumor	B-Cancer
environment	O
,	O
angiogenesis	O
and	O
,	O
particularly	O
,	O
vascular	B-Multi-tissue_structure
drainage	O
of	O
poorly	O
differentiated	O
neoplasms	B-Cancer
.	O

Syndecan	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
expression	O
in	O
mammary	B-Cell
epithelial	I-Cell
tumor	I-Cell
cells	I-Cell
is	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
-	O
dependent	O
.	O

E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
is	O
a	O
Ca	B-Simple_chemical
(	I-Simple_chemical
2	I-Simple_chemical
+	I-Simple_chemical
)	I-Simple_chemical
-	O
dependent	O
cell	B-Cell
-	O
cell	B-Cell
adhesion	O
molecule	O
,	O
which	O
is	O
mainly	O
expressed	O
in	O
epithelial	B-Cell
cells	I-Cell
.	O

Recent	O
studies	O
have	O
shown	O
that	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
has	O
an	O
important	O
role	O
as	O
an	O
invasion	O
suppressor	O
molecule	O
in	O
epithelial	B-Cell
tumor	I-Cell
cells	I-Cell
.	O

Syndecan	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
is	O
a	O
cell	B-Cellular_component
surface	I-Cellular_component
proteoglycan	O
that	O
has	O
been	O
implicated	O
in	O
a	O
number	O
of	O
cellular	B-Cell
functions	O
including	O
cell	B-Cell
-	O
cell	B-Cell
adhesion	O
,	O
cell	B-Cell
-	O
matrix	B-Cellular_component
anchorage	O
and	O
growth	B-Gene_or_gene_product
factor	I-Gene_or_gene_product
presentation	O
for	O
signalling	O
receptors	O
.	O

Its	O
suppression	O
has	O
also	O
been	O
shown	O
to	O
be	O
associated	O
with	O
malignant	O
transformation	O
of	O
epithelial	B-Cell
cells	I-Cell
.	O

In	O
order	O
to	O
better	O
understand	O
the	O
coordinated	O
regulation	O
of	O
cell	B-Cell
-	O
cell	B-Cell
and	O
cell	B-Cell
-	O
matrix	B-Cellular_component
interactions	O
during	O
malignant	O
transformation	O
,	O
we	O
have	O
studied	O
the	O
expression	O
of	O
syndecan	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
in	O
malignant	B-Cell
mammary	I-Cell
tumor	I-Cell
cells	I-Cell
genetically	O
manipulated	O
for	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
expression	O
.	O

In	O
invasive	O
NM	B-Cell
-	I-Cell
e	I-Cell
-	I-Cell
ras	I-Cell
-	I-Cell
MAC1	I-Cell
cells	I-Cell
,	O
where	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
was	O
partially	O
downregulated	O
by	O
specific	O
antisense	O
RNA	O
,	O
syndecan	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
expression	O
was	O
suppressed	O
.	O

Furthermore	O
,	O
transfection	O
of	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
cDNA	I-Gene_or_gene_product
into	O
invasive	O
NM	B-Cell
-	I-Cell
f	I-Cell
-	I-Cell
ras	I-Cell
-	I-Cell
TD	I-Cell
cells	I-Cell
resulted	O
in	O
the	O
upregulation	O
of	O
syndecan	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
expression	O
in	O
association	O
with	O
decreased	O
invasiveness	O
.	O

In	O
both	O
cases	O
,	O
regulation	O
of	O
syndecan	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
occurred	O
post	O
-	O
transcriptionally	O
,	O
since	O
syndecan	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
mRNA	O
levels	O
remained	O
unchanged	O
.	O

Instead	O
,	O
a	O
translational	O
regulation	O
is	O
suggested	O
,	O
since	O
syndecan	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
core	O
protein	O
synthesis	O
was	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
dependent	O
.	O

Another	O
cell	B-Cell
adhesion	O
protein	O
,	O
beta	B-Gene_or_gene_product
1	I-Gene_or_gene_product
-	I-Gene_or_gene_product
integrin	I-Gene_or_gene_product
was	O
not	O
affected	O
by	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
expression	O
.	O

The	O
data	O
provide	O
an	O
example	O
of	O
coordinated	O
changes	O
in	O
the	O
expression	O
of	O
two	O
cell	B-Cell
adhesion	O
molecules	O
,	O
syndecan	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
during	O
epithelial	B-Cell
cell	I-Cell
transformation	O
.	O

Metastasis	O
-	O
associated	O
5T4	B-Gene_or_gene_product
antigen	O
disrupts	O
cell	B-Cell
-	O
cell	B-Cell
contacts	O
and	O
induces	O
cellular	B-Cell
motility	O
in	O
epithelial	B-Cell
cells	I-Cell
.	O

The	O
5T4	B-Gene_or_gene_product
antigen	O
is	O
defined	O
by	O
a	O
monoclonal	O
antibody	O
(	O
MAb	O
)	O
specific	O
for	O
human	B-Organism
trophoblast	B-Cell
.	O

It	O
is	O
also	O
expressed	O
by	O
many	O
types	O
of	O
tumour	B-Cell
cell	I-Cell
and	O
has	O
been	O
associated	O
with	O
metastasis	O
and	O
poor	O
clinical	O
outcome	O
in	O
a	O
number	O
of	O
cancers	B-Cancer
.	O

This	O
pattern	O
of	O
expression	O
is	O
consistent	O
with	O
a	O
mechanistic	O
involvement	O
of	O
5T4	B-Gene_or_gene_product
molecules	O
in	O
the	O
spread	O
of	O
cancer	B-Cell
cells	I-Cell
.	O

The	O
5T4	B-Gene_or_gene_product
antigen	O
is	O
a	O
transmembrane	B-Cellular_component
glycoprotein	O
with	O
a	O
310	O
amino	B-Amino_acid
acid	I-Amino_acid
extracellular	B-Immaterial_anatomical_entity
domain	O
and	O
a	O
44	O
amino	B-Amino_acid
acid	I-Amino_acid
cytoplasmic	B-Organism_substance
domain	O
.	O

Transfection	O
of	O
full	O
-	O
length	O
5T4	B-Gene_or_gene_product
cDNA	O
into	O
epithelial	B-Cell
cells	I-Cell
alters	O
cell	B-Cell
-	O
cell	B-Cell
contacts	O
and	O
cellular	B-Cell
motility	O
.	O

Thus	O
,	O
in	O
5T4	B-Gene_or_gene_product
-	O
transfected	O
CL	B-Cell
-	I-Cell
S1	I-Cell
murine	I-Cell
mammary	I-Cell
cells	I-Cell
,	O
5T4	B-Gene_or_gene_product
expression	O
is	O
associated	O
with	O
dendritic	B-Cell
morphology	O
,	O
accompanied	O
by	O
abrogation	O
of	O
actin	B-Gene_or_gene_product
/	O
cadherin	B-Gene_or_gene_product
-	O
containing	O
contacts	O
and	O
increased	O
motility	O
.	O

In	O
transfected	O
MDCK	B-Cell
canine	I-Cell
kidney	I-Cell
epithelial	I-Cell
cells	I-Cell
,	O
5T4	B-Gene_or_gene_product
over	O
-	O
expression	O
also	O
results	O
in	O
increased	O
motility	O
,	O
but	O
disruption	O
of	O
cell	B-Cell
-	O
cell	B-Cell
contacts	O
,	O
either	O
by	O
culturing	O
cells	B-Cell
in	O
low	O
calcium	B-Simple_chemical
medium	O
or	O
by	O
addition	O
of	O
HGF	B-Gene_or_gene_product
/	O
SF	B-Gene_or_gene_product
,	O
is	O
needed	O
.	O

The	O
effects	O
of	O
5T4	B-Gene_or_gene_product
expression	O
on	O
morphology	O
and	O
motility	O
are	O
separable	O
since	O
cells	B-Cell
transfected	O
with	O
a	O
truncated	O
form	O
of	O
5T4	B-Gene_or_gene_product
cDNA	O
in	O
which	O
the	O
cytoplasmic	B-Organism_substance
domain	O
is	O
deleted	O
reveal	O
that	O
the	O
latter	O
is	O
necessary	O
to	O
abrogate	O
actin	B-Gene_or_gene_product
/	O
cadherin	B-Gene_or_gene_product
-	O
containing	O
contacts	O
but	O
does	O
not	O
influence	O
the	O
effects	O
on	O
motility	O
.	O

Thus	O
,	O
5T4	B-Gene_or_gene_product
molecules	O
can	O
deliver	O
signals	O
through	O
both	O
the	O
extracellular	B-Immaterial_anatomical_entity
and	O
intracellular	B-Immaterial_anatomical_entity
domains	O
,	O
and	O
the	O
resultant	O
effects	O
are	O
consistent	O
with	O
a	O
role	O
for	O
5T4	B-Gene_or_gene_product
molecules	O
in	O
invasion	O
processes	O
.	O

[	O
Effects	O
of	O
carcinostatic	O
agents	O
in	O
the	O
hematogenous	O
metastasis	O
of	O
cancer	B-Cancer
]	O
.	O

The	O
experimental	O
studies	O
in	O
vitro	O
and	O
in	O
vivo	O
were	O
performed	O
to	O
investigate	O
the	O
effects	O
of	O
carcinostatic	O
agents	O
in	O
the	O
adhesion	O
of	O
cancer	B-Cell
cells	I-Cell
to	O
endothelial	B-Cell
cells	I-Cell
.	O

Certain	O
carcinostatic	O
agents	O
induce	O
the	O
expression	O
of	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
selectin	I-Gene_or_gene_product
on	O
endothelial	B-Cell
cells	I-Cell
and	O
enhance	O
the	O
expression	O
of	O
carbohydrate	B-Simple_chemical
ligands	O
on	O
cancer	B-Cell
cells	I-Cell
.	O

Consequently	O
,	O
increased	O
adhesion	O
of	O
cancer	B-Cell
cells	I-Cell
to	O
endothelial	B-Cell
cells	I-Cell
was	O
observed	O
by	O
the	O
treatment	O
of	O
carcinostatic	O
agents	O
.	O

In	O
the	O
trans	B-Immaterial_anatomical_entity
-	I-Immaterial_anatomical_entity
splenic	I-Immaterial_anatomical_entity
hepatic	B-Organ
metastasis	O
model	O
using	O
nude	B-Organism
mice	I-Organism
,	O
the	O
augmentation	O
of	O
cancer	B-Cancer
metastasis	O
was	O
observed	O
by	O
the	O
treatment	O
of	O
carcinostatic	O
agents	O
.	O

From	O
the	O
above	O
,	O
we	O
concluded	O
that	O
carcinostatic	O
agents	O
may	O
increase	O
the	O
metastatic	O
ability	O
of	O
cancer	B-Cell
cells	I-Cell
in	O
patients	B-Organism
with	O
cancer	B-Cancer
.	O

Changes	O
in	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
levels	O
in	O
non	B-Cancer
-	I-Cancer
small	I-Cancer
cell	I-Cancer
lung	I-Cancer
cancer	I-Cancer
.	O

Increased	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
levels	O
are	O
increased	O
in	O
a	O
number	O
of	O
malignancies	B-Cancer
and	O
have	O
been	O
correlated	O
with	O
decreased	O
disease	O
-	O
free	O
interval	O
and	O
decreased	O
overall	O
survival	O
.	O

We	O
have	O
,	O
therefore	O
,	O
examined	O
components	O
of	O
this	O
plasminogen	B-Gene_or_gene_product
activating	O
system	O
in	O
patients	B-Organism
with	O
Non	B-Cancer
-	I-Cancer
Small	I-Cancer
Cell	I-Cancer
Lung	I-Cancer
Cancer	I-Cancer
(	O
NSCLC	B-Cancer
)	O
.	O

Levels	O
of	O
uPA	B-Gene_or_gene_product
,	O
urokinase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
plasminogen	I-Gene_or_gene_product
activator	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
(	O
uPAR	B-Gene_or_gene_product
)	O
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
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
were	O
measured	O
semiquantitatively	O
in	O
paraffin	O
sections	B-Cancer
of	O
tumours	B-Cancer
from	O
147	O
patients	B-Organism
with	O
NSCLC	B-Cancer
.	O

Immunohistochemically	O
stained	O
sections	B-Cancer
of	O
tumour	B-Cancer
were	O
allocated	O
a	O
score	O
for	O
stain	O
intensity	O
and	O
results	O
correlated	O
to	O
:	O
survival	O
;	O
tumour	B-Cancer
stage	O
(	O
T	O
)	O
;	O
nodal	B-Multi-tissue_structure
stage	O
(	O
N	O
)	O
;	O
stage	O
grouping	O
(	O
I	O
to	O
IIIb	O
)	O
,	O
survival	O
status	O
and	O
sex	O
.	O

Increased	O
levels	O
of	O
PAI	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
were	O
associated	O
with	O
a	O
decreased	O
survival	O
in	O
squamous	B-Cancer
cell	I-Cancer
carcinoma	I-Cancer
(	O
SCC	B-Cancer
)	O
X2	O
=	O
5	O
.	O
72	O
,	O
p	O
=	O
0	O
.	O
017	O
(	O
n	O
=	O
74	O
)	O
.	O

There	O
was	O
a	O
significant	O
positive	O
relationship	O
between	O
PAI	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
levels	O
and	O
N	O
-	O
stage	O
(	O
p	O
=	O
<	O
0	O
.	O
05	O
)	O
,	O
presence	O
of	O
nodal	B-Multi-tissue_structure
metastases	O
(	O
p	O
=	O
<	O
0	O
.	O
05	O
)	O
,	O
stage	O
grouping	O
(	O
p	O
=	O
<	O
0	O
.	O
01	O
)	O
and	O
extent	O
of	O
disease	O
(	O
p	O
=	O
<	O
0	O
.	O
05	O
)	O
in	O
the	O
total	O
group	O
and	O
the	O
SCC	B-Cancer
subgroup	O
,	O
but	O
not	O
adenocarcinoma	B-Cancer
.	O

There	O
was	O
a	O
significant	O
positive	O
relationship	O
between	O
PAI	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
levels	O
and	O
T	O
-	O
stage	O
(	O
p	O
=	O
<	O
0	O
.	O
05	O
)	O
in	O
the	O
total	O
group	O
,	O
and	O
survival	O
status	O
(	O
p	O
=	O
<	O
0	O
.	O
05	O
)	O
in	O
the	O
SCC	B-Cancer
subgroup	O
alone	O
.	O

uPA	B-Gene_or_gene_product
and	O
uPAR	B-Gene_or_gene_product
levels	O
were	O
not	O
significantly	O
associated	O
with	O
tumour	B-Cancer
staging	O
or	O
survival	O
.	O

We	O
conclude	O
that	O
increased	O
PAI	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
antigen	O
levels	O
may	O
be	O
associated	O
with	O
a	O
decreased	O
survival	O
in	O
patients	B-Organism
with	O
SCC	B-Cancer
.	O

Potential	O
roles	O
for	O
focal	B-Gene_or_gene_product
adhesion	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
in	O
development	O
.	O

Focal	B-Gene_or_gene_product
adhesion	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
(	O
pp125FAK	B-Gene_or_gene_product
or	O
FAK	B-Gene_or_gene_product
)	O
is	O
a	O
protein	B-Gene_or_gene_product
tyrosine	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
which	O
is	O
associated	O
with	O
intracellular	B-Immaterial_anatomical_entity
signalling	O
cascades	O
which	O
are	O
initiated	O
when	O
the	O
integrin	B-Gene_or_gene_product
family	O
of	O
cell	B-Gene_or_gene_product
adhesion	I-Gene_or_gene_product
molecules	I-Gene_or_gene_product
engage	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
molecules	O
.	O

In	O
cultured	O
cells	B-Cell
,	O
this	O
molecule	O
is	O
physically	O
associated	O
with	O
focal	B-Cellular_component
adhesions	I-Cellular_component
,	O
which	O
are	O
well	O
-	O
defined	O
regions	O
of	O
intimate	O
cell	B-Cell
-	O
to	O
-	O
substratum	O
adhesion	O
.	O

In	O
this	O
location	O
,	O
it	O
interacts	O
with	O
other	O
proteins	O
of	O
the	O
focal	B-Cellular_component
adhesion	I-Cellular_component
to	O
activate	O
intracellular	B-Immaterial_anatomical_entity
signalling	O
events	O
associated	O
with	O
cell	B-Cell
adhesion	O
.	O

The	O
in	O
vitro	O
expression	O
of	O
FAK	B-Gene_or_gene_product
and	O
its	O
level	O
of	O
phosphorylation	O
appear	O
to	O
be	O
related	O
to	O
several	O
physiological	O
phenomena	O
,	O
including	O
cell	B-Cell
spreading	O
,	O
cell	B-Cell
differentiation	O
,	O
cell	B-Cell
locomotion	O
and	O
cell	B-Cell
death	O
.	O

Because	O
these	O
phenomena	O
are	O
all	O
of	O
critical	O
importance	O
during	O
morphogenesis	O
,	O
and	O
because	O
FAK	B-Gene_or_gene_product
is	O
expressed	O
in	O
embryonic	B-Cell
cells	I-Cell
,	O
evidence	O
has	O
been	O
accumulating	O
to	O
indicate	O
that	O
FAK	B-Gene_or_gene_product
may	O
be	O
an	O
important	O
modulator	O
of	O
developmental	O
processes	O
.	O

In	O
this	O
review	O
,	O
this	O
evidence	O
is	O
surveyed	O
together	O
with	O
evidence	O
from	O
analogous	O
situations	O
,	O
such	O
as	O
tumour	B-Cell
cell	I-Cell
migration	O
and	O
invasiveness	O
.	O

Although	O
evidence	O
suggesting	O
a	O
role	O
for	O
FAK	B-Gene_or_gene_product
in	O
morphogenesis	O
is	O
accumulating	O
,	O
current	O
uncertainties	O
regarding	O
its	O
cytoplasmic	B-Organism_substance
location	O
and	O
its	O
molecular	O
interactions	O
in	O
vivo	O
make	O
it	O
difficult	O
to	O
reach	O
definitive	O
conclusions	O
regarding	O
the	O
significance	O
of	O
its	O
contributions	O
to	O
developmental	O
processes	O
.	O

