Positron	O
emission	O
tomography	O
in	O
a	O
case	O
of	O
intracranial	B-Cancer
hemangiopericytoma	I-Cancer
.	O

Due	O
to	O
the	O
low	O
prevalence	O
of	O
hemangiopericytomas	B-Cancer
(	O
HPCs	B-Cancer
)	O
,	O
data	O
on	O
the	O
biophysiological	O
characteristics	O
of	O
this	O
tumor	B-Cancer
are	O
rare	O
.	O

Positron	O
emission	O
tomography	O
(	O
PET	O
)	O
demonstrated	O
a	O
sixfold	O
increased	O
uptake	O
of	O
[	B-Simple_chemical
11C	I-Simple_chemical
]	I-Simple_chemical
methionine	I-Simple_chemical
and	O
hyperperfusion	O
in	O
the	O
HPC	B-Cancer
,	O
whereas	O
glucose	B-Simple_chemical
utilization	O
was	O
decreased	O
in	O
this	O
area	O
.	O

This	O
low	O
glucose	B-Simple_chemical
utilization	O
is	O
in	O
contrast	O
to	O
the	O
high	O
[	B-Simple_chemical
11C	I-Simple_chemical
]	I-Simple_chemical
methionine	I-Simple_chemical
uptake	O
and	O
the	O
malignancy	O
of	O
these	O
tumors	B-Cancer
.	O

The	O
characteristics	O
of	O
HPCs	B-Cancer
in	O
PET	O
described	O
herein	O
for	O
the	O
first	O
time	O
offer	O
additional	O
diagnostic	O
criteria	O
and	O
may	O
help	O
especially	O
to	O
differentiate	O
these	O
tumors	B-Cancer
from	O
meningiomas	B-Cancer
.	O

Thyrotropin	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
:	O
a	O
role	O
for	O
thyroid	B-Organ
tumourigenesis	O
?	O

Human	B-Organism
thyroid	B-Cancer
tumours	I-Cancer
represent	O
an	O
example	O
of	O
the	O
interplay	O
of	O
genetic	O
and	O
non	O
genetic	O
carcinogenesis	O
.	O

Recently	O
,	O
genetic	O
abnormalities	O
in	O
the	O
elements	O
of	O
the	O
Thyrotropin	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
(	O
TSH	B-Gene_or_gene_product
-	I-Gene_or_gene_product
R	I-Gene_or_gene_product
)	O
dependent	O
cAMP	B-Simple_chemical
regulatory	O
cascade	O
have	O
been	O
found	O
to	O
be	O
involved	O
both	O
in	O
benign	B-Pathological_formation
and	O
malignant	B-Cancer
thyroid	I-Cancer
tumours	I-Cancer
.	O

The	O
presence	O
of	O
activating	O
mutations	O
has	O
been	O
demonstrated	O
in	O
the	O
TSH	B-Gene_or_gene_product
-	I-Gene_or_gene_product
R	I-Gene_or_gene_product
gene	O
as	O
well	O
as	O
in	O
the	O
Gs	B-Gene_or_gene_product
alpha	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
gene	O
in	O
thyroid	B-Cancer
toxic	I-Cancer
adenoma	I-Cancer
resulting	O
in	O
the	O
constitutive	O
activation	O
of	O
the	O
cAMP	B-Simple_chemical
pathway	O
and	O
it	O
has	O
been	O
hypothesised	O
that	O
these	O
genetic	O
alterations	O
may	O
play	O
a	O
causative	O
role	O
in	O
the	O
disease	O
.	O

However	O
,	O
recent	O
observations	O
suggest	O
more	O
caution	O
in	O
accepting	O
such	O
a	O
hypothesis	O
.	O

The	O
presence	O
of	O
activating	O
TSH	B-Gene_or_gene_product
-	I-Gene_or_gene_product
R	I-Gene_or_gene_product
mutations	O
has	O
also	O
been	O
demonstrated	O
in	O
differentiated	O
thyroid	B-Cancer
carcinomas	I-Cancer
.	O

At	O
present	O
,	O
the	O
percentage	O
of	O
such	O
a	O
modification	O
is	O
low	O
,	O
unless	O
referred	O
to	O
selected	O
series	O
of	O
tumours	B-Cancer
.	O

Activating	O
mutations	O
of	O
the	O
TSH	B-Gene_or_gene_product
-	I-Gene_or_gene_product
R	I-Gene_or_gene_product
gene	O
have	O
been	O
detected	O
in	O
a	O
group	O
of	O
differentiated	O
carcinomas	B-Cancer
with	O
high	O
basal	O
adenylyl	B-Gene_or_gene_product
cyclase	I-Gene_or_gene_product
activity	O
,	O
and	O
in	O
a	O
few	O
cases	O
of	O
hyperfunctioning	O
thyroid	B-Cancer
carcinoma	I-Cancer
.	O

However	O
,	O
the	O
role	O
of	O
the	O
TSH	B-Gene_or_gene_product
-	I-Gene_or_gene_product
R	I-Gene_or_gene_product
-	I-Gene_or_gene_product
related	O
cAMP	B-Simple_chemical
pathway	O
alterations	O
in	O
thyroid	B-Organ
transformation	O
remains	O
to	O
be	O
elucidated	O
.	O

In	O
this	O
review	O
,	O
the	O
role	O
of	O
TSH	B-Gene_or_gene_product
-	I-Gene_or_gene_product
R	I-Gene_or_gene_product
gene	O
alterations	O
in	O
benign	B-Pathological_formation
and	O
malignant	B-Cancer
thyroid	I-Cancer
neoplasia	I-Cancer
is	O
examined	O
.	O

Cooccurrence	O
of	O
reduced	O
expression	O
of	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
and	O
overexpression	O
of	O
p53	B-Gene_or_gene_product
is	O
a	O
predictor	O
of	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
in	O
early	O
gastric	B-Cancer
cancer	I-Cancer
.	O

BACKGROUND	O
AND	O
OBJECTIVES	O
:	O
Even	O
though	O
the	O
pathological	O
background	O
contributes	O
to	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
,	O
the	O
biological	O
characteristics	O
of	O
tumors	B-Cancer
have	O
also	O
gained	O
wide	O
attention	O
.	O

In	O
this	O
study	O
,	O
the	O
expression	O
of	O
the	O
cadherin	B-Gene_or_gene_product
-	O
catenin	B-Gene_or_gene_product
complex	O
and	O
p53	B-Gene_or_gene_product
was	O
studied	O
in	O
early	O
gastric	B-Cancer
cancer	I-Cancer
.	O

Their	O
correlation	O
with	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
and	O
the	O
predictability	O
of	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastases	O
,	O
by	O
combining	O
these	O
factors	O
,	O
were	O
also	O
discussed	O
.	O

METHODS	O
:	O
One	O
hundred	O
and	O
one	O
specimens	B-Multi-tissue_structure
obtained	O
from	O
surgery	O
were	O
studied	O
by	O
immunohistochemistry	O
using	O
monoclonal	O
anti	O
-	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
,	O
anti	O
-	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
and	O
anti	O
-	O
p53	B-Gene_or_gene_product
antibodies	O
.	O

RESULTS	O
:	O
Expression	O
of	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
and	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
was	O
reduced	O
in	O
50	O
.	O
5	O
and	O
64	O
.	O
4	O
%	O
,	O
respectively	O
.	O

p53	B-Gene_or_gene_product
protein	O
staining	O
was	O
positive	O
in	O
29	O
.	O
7	O
%	O
.	O

There	O
was	O
a	O
significant	O
correlation	O
between	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
and	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
expression	O
,	O
but	O
no	O
correlation	O
was	O
found	O
between	O
p53	B-Gene_or_gene_product
expression	O
and	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
or	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
expression	O
.	O

A	O
reduction	O
in	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
expression	O
and	O
p53	B-Gene_or_gene_product
overexpression	O
correlated	O
to	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastases	O
,	O
respectively	O
.	O

Multivariate	O
analysis	O
showed	O
that	O
cooccurrence	O
of	O
reduced	O
expression	O
of	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
and	O
overexpression	O
of	O
p53	B-Gene_or_gene_product
was	O
an	O
independent	O
factor	O
indicating	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastases	O
.	O

CONCLUSION	O
:	O
A	O
study	O
of	O
both	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
and	O
p53	B-Gene_or_gene_product
expression	O
may	O
be	O
helpful	O
to	O
predict	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastases	O
in	O
early	O
gastric	B-Cancer
cancer	I-Cancer
.	O

Proto	O
-	O
oncogene	O
N	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
promoter	O
is	O
down	O
regulated	O
by	O
the	O
Wilms	B-Cancer
'	I-Cancer
tumor	I-Cancer
suppressor	O
gene	O
WT1	B-Gene_or_gene_product
.	O

The	O
Wilms	B-Gene_or_gene_product
'	I-Gene_or_gene_product
tumor	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
WT1	B-Gene_or_gene_product
)	O
gene	O
is	O
a	O
tumor	B-Cancer
suppressor	O
gene	O
that	O
encodes	O
a	O
zinc	B-Simple_chemical
-	O
finger	O
transcription	O
factor	O
.	O

WT1	B-Gene_or_gene_product
represses	O
transcription	O
of	O
several	O
growth	O
factors	O
and	O
growth	O
factor	O
receptors	O
.	O

The	O
N	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
proto	O
-	O
oncogene	O
encodes	O
a	O
transcription	O
factor	O
which	O
regulates	O
cell	B-Cell
growth	O
and	O
differentiation	O
.	O

N	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
is	O
coexpressed	O
with	O
WT1	B-Gene_or_gene_product
in	O
the	O
developing	O
kidney	B-Organ
and	O
is	O
overexpressed	O
in	O
many	O
Wilms	B-Cancer
'	I-Cancer
tumors	I-Cancer
.	O

Here	O
,	O
we	O
show	O
that	O
the	O
proto	O
-	O
oncogene	O
N	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
promoter	O
was	O
down	O
-	O
regulated	O
by	O
WT1	B-Gene_or_gene_product
in	O
transient	O
transfection	O
assays	O
.	O

However	O
,	O
mutant	O
WT1	B-Gene_or_gene_product
(	O
R394W	B-Amino_acid
)	O
which	O
has	O
a	O
mutation	O
in	O
the	O
DNA	B-Cellular_component
binding	O
domain	O
could	O
not	O
repress	O
the	O
N	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
promoter	O
.	O

Electrophoretic	O
mobility	O
shift	O
assays	O
showed	O
that	O
the	O
oligonucleotides	O
containing	O
the	O
WT1	B-Gene_or_gene_product
motifs	O
could	O
bind	O
recombinant	O
WT1	B-Gene_or_gene_product
proteins	O
.	O

This	O
suggests	O
that	O
the	O
repression	O
of	O
the	O
N	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
promoter	O
is	O
mediated	O
through	O
the	O
WT1	B-Gene_or_gene_product
binding	O
sites	O
.	O

This	O
finding	O
may	O
help	O
to	O
elucidate	O
the	O
relationship	O
of	O
WT1	B-Gene_or_gene_product
and	O
N	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
in	O
tumorigenesis	O
and	O
renal	B-Organ
development	O
.	O

Tumour	B-Cancer
-	O
specific	O
distribution	O
of	O
BRCA1	B-Gene_or_gene_product
promoter	O
region	O
methylation	O
supports	O
a	O
pathogenetic	O
role	O
in	O
breast	B-Cancer
and	O
ovarian	B-Cancer
cancer	I-Cancer
.	O

The	O
role	O
of	O
BRCA1	B-Gene_or_gene_product
in	O
sporadic	O
breast	B-Cancer
and	O
ovarian	B-Cancer
cancers	I-Cancer
remains	O
elusive	O
.	O

Direct	O
involvement	O
of	O
BRCA1	B-Gene_or_gene_product
in	O
the	O
development	O
of	O
breast	B-Cancer
and	O
ovarian	B-Cancer
cancer	I-Cancer
is	O
suggested	O
by	O
the	O
finding	O
that	O
the	O
BRCA1	B-Gene_or_gene_product
promoter	O
region	O
CpG	O
island	O
is	O
methylated	O
in	O
a	O
proportion	O
of	O
breast	B-Cancer
and	O
ovarian	B-Cancer
cancers	I-Cancer
.	O

The	O
aim	O
of	O
this	O
study	O
was	O
to	O
compare	O
the	O
incidence	O
of	O
BRCA1	B-Gene_or_gene_product
promoter	O
region	O
methylation	O
in	O
tumours	B-Cancer
in	O
which	O
loss	O
of	O
BRCA1	B-Gene_or_gene_product
has	O
been	O
shown	O
to	O
play	O
a	O
role	O
in	O
pathogenesis	O
(	O
breast	B-Cancer
and	O
ovarian	B-Cancer
carcinomas	I-Cancer
)	O
with	O
the	O
incidence	O
in	O
tumours	B-Cancer
in	O
which	O
BRCA1	B-Gene_or_gene_product
is	O
unlikely	O
to	O
play	O
a	O
role	O
in	O
pathogenesis	O
.	O

Promoter	O
region	O
hypermethylation	O
was	O
significantly	O
more	O
common	O
(	O
P	O
<	O
0	O
.	O
008	O
)	O
in	O
breast	B-Cancer
and	O
ovarian	B-Cancer
cancer	I-Cancer
(	O
6	O
/	O
38	O
tumours	B-Cancer
methylated	O
)	O
than	O
in	O
colon	B-Cancer
cancer	I-Cancer
(	O
0	O
/	O
35	O
tumours	B-Cancer
methylated	O
)	O
or	O
in	O
leukaemias	B-Cancer
(	O
0	O
/	O
19	O
samples	B-Cancer
methylated	O
)	O
.	O

The	O
restriction	O
of	O
BRCA1	B-Gene_or_gene_product
promoter	O
region	O
hypermethylation	O
to	O
breast	B-Cancer
and	O
ovarian	B-Cancer
cancer	I-Cancer
is	O
consistent	O
with	O
a	O
pathogenetic	O
role	O
of	O
BRCA1	B-Gene_or_gene_product
promoter	O
methylation	O
in	O
these	O
tumours	B-Cancer
.	O

We	O
suggest	O
that	O
the	O
rarity	O
of	O
observed	O
BRCA1	B-Gene_or_gene_product
mutations	O
in	O
sporadic	O
breast	B-Cancer
and	O
ovarian	B-Cancer
cancer	I-Cancer
is	O
due	O
to	O
the	O
greater	O
likelihood	O
of	O
BRCA1	B-Gene_or_gene_product
inactivation	O
by	O
non	O
-	O
mutational	O
mechanisms	O
such	O
as	O
methylation	O
.	O

Regulation	O
of	O
the	O
resident	O
chromosomal	B-Cellular_component
copy	O
of	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
by	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Myb	I-Gene_or_gene_product
is	O
involved	O
in	O
myeloid	B-Cell
leukemogenesis	O
.	O

c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myb	I-Gene_or_gene_product
is	O
a	O
frequent	O
target	O
of	O
retroviral	B-Organism
insertional	O
mutagenesis	O
in	O
murine	B-Organism
leukemia	I-Organism
virus	I-Organism
-	O
induced	O
myeloid	B-Cancer
leukemia	I-Cancer
.	O

Induction	O
of	O
the	O
leukemogenic	O
phenotype	O
is	O
generally	O
associated	O
with	O
inappropriate	O
expression	O
of	O
this	O
transcriptional	O
regulator	O
.	O

Despite	O
intensive	O
investigations	O
,	O
the	O
target	O
genes	O
of	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myb	I-Gene_or_gene_product
that	O
are	O
specifically	O
involved	O
in	O
development	O
of	O
these	O
myeloid	B-Cell
lineage	O
neoplasms	B-Cancer
are	O
still	O
unknown	O
.	O

In	O
vitro	O
assays	O
have	O
indicated	O
that	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
may	O
be	O
a	O
target	O
gene	O
of	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Myb	I-Gene_or_gene_product
;	O
however	O
,	O
regulation	O
of	O
the	O
resident	O
chromosomal	B-Cellular_component
gene	O
has	O
not	O
yet	O
been	O
demonstrated	O
.	O

To	O
address	O
this	O
question	O
further	O
,	O
we	O
analyzed	O
the	O
expression	O
of	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
in	O
a	O
myeloblastic	B-Cell
cell	I-Cell
line	I-Cell
,	O
M1	B-Cell
,	O
expressing	O
a	O
conditionally	O
active	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Myb	I-Gene_or_gene_product
-	I-Gene_or_gene_product
estrogen	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
fusion	O
protein	O
(	O
MybER	B-Gene_or_gene_product
)	O
.	O

Activation	O
of	O
MybER	B-Gene_or_gene_product
both	O
prevented	O
the	O
growth	O
arrest	O
induced	O
by	O
interleukin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
6	I-Gene_or_gene_product
(	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
6	I-Gene_or_gene_product
)	O
and	O
rapidly	O
restored	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
expression	O
in	O
nearly	O
terminal	O
differentiated	O
cells	B-Cell
that	O
had	O
been	O
exposed	O
to	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
6	I-Gene_or_gene_product
for	O
3	O
days	O
.	O

Restoration	O
occurred	O
in	O
the	O
presence	O
of	O
a	O
protein	O
synthesis	O
inhibitor	O
but	O
not	O
after	O
a	O
transcriptional	O
block	O
,	O
indicating	O
that	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
is	O
a	O
direct	O
,	O
transcriptionally	O
regulated	O
target	O
of	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Myb	I-Gene_or_gene_product
.	O

c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
is	O
a	O
major	O
target	O
that	O
transduces	O
Myb	B-Gene_or_gene_product
'	O
s	O
proliferative	O
signal	O
,	O
as	O
shown	O
by	O
the	O
ability	O
of	O
a	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Myc	I-Gene_or_gene_product
-	I-Gene_or_gene_product
estrogen	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
fusion	O
protein	O
alone	O
to	O
also	O
reverse	O
growth	O
arrest	O
in	O
this	O
system	O
.	O

To	O
investigate	O
the	O
possibility	O
that	O
this	O
regulatory	O
connection	O
contributes	O
to	O
Myb	B-Gene_or_gene_product
'	O
s	O
oncogenicity	O
,	O
we	O
expressed	O
a	O
dominant	O
negative	O
Myb	B-Gene_or_gene_product
in	O
the	O
myeloid	B-Cell
leukemic	I-Cell
cell	I-Cell
line	I-Cell
RI	I-Cell
-	I-Cell
4	I-Cell
-	I-Cell
11	I-Cell
.	O

In	O
this	O
cell	B-Cell
line	I-Cell
,	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myb	I-Gene_or_gene_product
is	O
activated	O
by	O
insertional	O
mutagenesis	O
and	O
cannot	O
be	O
effectively	O
down	O
regulated	O
by	O
cytokine	O
.	O

Myb	B-Gene_or_gene_product
'	O
s	O
ability	O
to	O
regulate	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
'	I-Gene_or_gene_product
s	I-Gene_or_gene_product
expression	O
was	O
also	O
demonstrated	O
in	O
these	O
cells	B-Cell
,	O
showing	O
a	O
mechanism	O
through	O
which	O
the	O
proto	O
-	O
oncogene	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myb	I-Gene_or_gene_product
can	O
exert	O
its	O
oncogenic	O
potential	O
in	O
myeloid	B-Cell
lineage	I-Cell
hematopoietic	I-Cell
cells	I-Cell
.	O

Adenovirus	B-Organism
-	O
mediated	O
gene	O
transfer	O
of	O
endostatin	B-Gene_or_gene_product
in	O
vivo	O
results	O
in	O
high	O
level	O
of	O
transgene	O
expression	O
and	O
inhibition	O
of	O
tumor	B-Cancer
growth	O
and	O
metastases	O
.	O

Inhibition	O
of	O
angiogenesis	O
has	O
been	O
shown	O
to	O
be	O
an	O
effective	O
strategy	O
in	O
cancer	B-Cancer
therapy	O
in	O
mice	B-Organism
.	O

However	O
,	O
its	O
widespread	O
application	O
has	O
been	O
hampered	O
by	O
difficulties	O
in	O
the	O
large	O
-	O
scale	O
production	O
of	O
the	O
antiangiogenic	O
proteins	O
.	O

This	O
limitation	O
may	O
be	O
resolved	O
by	O
in	O
vivo	O
delivery	O
and	O
expression	O
of	O
the	O
antiangiogenic	O
genes	O
.	O

We	O
have	O
constructed	O
a	O
recombinant	O
adenovirus	B-Organism
that	O
expresses	O
murine	B-Organism
endostatin	B-Gene_or_gene_product
that	O
is	O
biologically	O
active	O
both	O
in	O
vitro	O
,	O
as	O
determined	O
in	O
endothelial	B-Cell
cell	I-Cell
proliferation	O
assays	O
,	O
and	O
in	O
vivo	O
,	O
by	O
suppression	O
of	O
angiogenesis	O
induced	O
by	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
165	I-Gene_or_gene_product
.	O

Persistent	O
high	O
serum	B-Organism_substance
levels	O
of	O
endostatin	B-Gene_or_gene_product
(	O
605	O
-	O
1740	O
ng	O
/	O
ml	O
;	O
mean	O
,	O
936	O
ng	O
/	O
ml	O
)	O
were	O
achieved	O
after	O
systemic	O
administration	O
of	O
the	O
vector	O
to	O
nude	B-Organism
mice	I-Organism
,	O
which	O
resulted	O
in	O
significant	O
reduction	O
of	O
the	O
growth	O
rates	O
and	O
the	O
volumes	O
of	O
JC	B-Cancer
breast	I-Cancer
carcinoma	I-Cancer
and	O
Lewis	B-Cancer
lung	I-Cancer
carcinoma	I-Cancer
(	O
P	O
less	O
than	O
0	O
.	O
001	O
and	O
P	O
less	O
than	O
0	O
.	O
05	O
,	O
respectively	O
)	O
.	O

In	O
addition	O
,	O
the	O
endostatin	B-Gene_or_gene_product
vector	O
treatment	O
completely	O
prevented	O
the	O
formation	O
of	O
pulmonary	B-Cancer
micrometastases	I-Cancer
in	O
Lewis	B-Cancer
lung	I-Cancer
carcinoma	I-Cancer
(	O
P	O
=	O
0	O
.	O
0001	O
)	O
.	O

Immunohistochemical	O
staining	O
of	O
the	O
tumors	B-Cancer
demonstrated	O
a	O
decreased	O
number	O
of	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
in	O
the	O
treatment	O
group	O
versus	O
the	O
controls	O
.	O

In	O
conclusion	O
,	O
the	O
present	O
study	O
clearly	O
demonstrates	O
the	O
potential	O
of	O
vector	O
-	O
mediated	O
antiangiogenic	O
gene	O
therapy	O
as	O
a	O
component	O
in	O
cancer	B-Cancer
therapy	O
.	O

High	O
frequency	O
of	O
hypermethylation	O
at	O
the	O
14	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
sigma	I-Gene_or_gene_product
locus	O
leads	O
to	O
gene	O
silencing	O
in	O
breast	B-Cancer
cancer	I-Cancer
.	O

Expression	O
of	O
14	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
final	I-Gene_or_gene_product
sigma	I-Gene_or_gene_product
(	O
final	B-Gene_or_gene_product
sigma	I-Gene_or_gene_product
)	O
is	O
induced	O
in	O
response	O
to	O
DNA	B-Cellular_component
damage	O
,	O
and	O
causes	O
cells	B-Cell
to	O
arrest	O
in	O
G	O
(	O
2	O
)	O
.	O

By	O
SAGE	O
(	O
serial	O
analysis	O
of	O
gene	O
expression	O
)	O
analysis	O
,	O
we	O
identified	O
final	B-Gene_or_gene_product
sigma	I-Gene_or_gene_product
as	O
a	O
gene	O
whose	O
expression	O
is	O
7	O
-	O
fold	O
lower	O
in	O
breast	B-Cell
carcinoma	I-Cell
cells	I-Cell
than	O
in	O
normal	O
breast	B-Tissue
epithelium	I-Tissue
.	O

We	O
verified	O
this	O
finding	O
by	O
Northern	O
blot	O
analysis	O
.	O

Remarkably	O
,	O
final	B-Gene_or_gene_product
sigma	I-Gene_or_gene_product
mRNA	O
was	O
undetectable	O
in	O
45	O
of	O
48	O
primary	O
breast	B-Cancer
carcinomas	I-Cancer
.	O

Genetic	O
alterations	O
at	O
final	B-Gene_or_gene_product
sigma	I-Gene_or_gene_product
such	O
as	O
loss	O
of	O
heterozygosity	O
were	O
rare	O
(	O
1	O
/	O
20	O
informative	O
cases	O
)	O
,	O
and	O
no	O
mutations	O
were	O
detected	O
(	O
0	O
/	O
34	O
)	O
.	O

On	O
the	O
other	O
hand	O
,	O
hypermethylation	O
of	O
CpG	O
islands	O
in	O
the	O
final	B-Gene_or_gene_product
sigma	I-Gene_or_gene_product
gene	O
was	O
detected	O
in	O
91	O
%	O
(	O
75	O
/	O
82	O
)	O
of	O
breast	B-Cancer
tumors	I-Cancer
and	O
was	O
associated	O
with	O
lack	O
of	O
gene	O
expression	O
.	O

Hypermethylation	O
of	O
final	B-Gene_or_gene_product
sigma	I-Gene_or_gene_product
is	O
functionally	O
important	O
,	O
because	O
treatment	O
of	O
final	B-Gene_or_gene_product
sigma	I-Gene_or_gene_product
-	O
non	O
-	O
expressing	O
breast	B-Cell
cancer	I-Cell
cell	I-Cell
lines	I-Cell
with	O
the	O
drug	O
5	B-Simple_chemical
-	I-Simple_chemical
aza	I-Simple_chemical
-	I-Simple_chemical
2	I-Simple_chemical
'	I-Simple_chemical
-	I-Simple_chemical
deoxycytidine	I-Simple_chemical
resulted	O
in	O
demethylation	O
of	O
the	O
gene	O
and	O
synthesis	O
of	O
final	B-Gene_or_gene_product
sigma	I-Gene_or_gene_product
mRNA	O
.	O

Breast	B-Cell
cancer	I-Cell
cells	I-Cell
lacking	O
final	B-Gene_or_gene_product
sigma	I-Gene_or_gene_product
expression	O
showed	O
increased	O
number	O
of	O
chromosomal	B-Cellular_component
breaks	O
and	O
gaps	O
when	O
exposed	O
to	O
gamma	O
-	O
irradiation	O
.	O

Therefore	O
,	O
it	O
is	O
possible	O
that	O
loss	O
of	O
final	B-Gene_or_gene_product
sigma	I-Gene_or_gene_product
expression	O
contributes	O
to	O
malignant	O
transformation	O
by	O
impairing	O
the	O
G	O
(	O
2	O
)	O
cell	B-Cell
cycle	O
checkpoint	O
function	O
,	O
thus	O
allowing	O
an	O
accumulation	O
of	O
genetic	O
defects	O
.	O

Hypermethylation	O
and	O
loss	O
of	O
final	B-Gene_or_gene_product
sigma	I-Gene_or_gene_product
expression	O
are	O
the	O
most	O
consistent	O
molecular	O
alterations	O
in	O
breast	B-Cancer
cancer	I-Cancer
identified	O
so	O
far	O
.	O

Reduced	O
oncogenicity	O
of	O
p190	B-Gene_or_gene_product
Bcr	I-Gene_or_gene_product
/	I-Gene_or_gene_product
Abl	I-Gene_or_gene_product
F	B-Gene_or_gene_product
-	I-Gene_or_gene_product
actin	I-Gene_or_gene_product
-	O
binding	O
domain	O
mutants	O
.	O

The	O
deregulated	O
Bcr	B-Gene_or_gene_product
/	I-Gene_or_gene_product
Abl	I-Gene_or_gene_product
tyrosine	B-Amino_acid
kinase	O
is	O
responsible	O
for	O
the	O
development	O
of	O
Philadelphia	O
(	O
Ph	O
)	O
-	O
positive	O
leukemia	B-Cancer
in	O
humans	B-Organism
.	O

To	O
investigate	O
the	O
significance	O
of	O
the	O
C	O
-	O
terminal	O
Abl	B-Gene_or_gene_product
actin	B-Gene_or_gene_product
-	O
binding	O
domain	O
within	O
Bcr	B-Gene_or_gene_product
/	I-Gene_or_gene_product
Abl	I-Gene_or_gene_product
p190	I-Gene_or_gene_product
in	O
the	O
development	O
of	O
leukemia	B-Cancer
/	O
lymphoma	B-Cancer
in	O
vivo	O
,	O
mutant	O
p190	B-Gene_or_gene_product
DNA	B-Cellular_component
constructs	O
were	O
used	O
to	O
generate	O
transgenic	B-Organism
mice	I-Organism
.	O

Eight	O
founder	O
and	O
progeny	O
mice	B-Organism
of	O
5	O
different	O
lines	O
were	O
monitored	O
for	O
leukemogenesis	O
.	O

Latency	O
was	O
markedly	O
increased	O
and	O
occurrence	O
decreased	O
in	O
the	O
p190	B-Gene_or_gene_product
del	O
C	O
lines	B-Organism
as	O
compared	O
with	O
nonmutated	O
p190	B-Gene_or_gene_product
BCR	I-Gene_or_gene_product
/	I-Gene_or_gene_product
ABL	I-Gene_or_gene_product
transgenics	B-Organism
.	O

Western	O
blot	O
analysis	O
of	O
involved	O
hematologic	B-Tissue
tissues	I-Tissue
of	O
the	O
p190	B-Gene_or_gene_product
del	O
C	O
transgenics	B-Organism
with	O
end	O
-	O
stage	O
disease	O
showed	O
high	O
-	O
level	O
expression	O
of	O
the	O
transgene	O
and	O
tyrosine	B-Amino_acid
phosphorylation	O
of	O
Cbl	B-Gene_or_gene_product
and	O
Hef1	B-Gene_or_gene_product
/	O
Cas	B-Gene_or_gene_product
,	O
proteins	O
previously	O
shown	O
to	O
be	O
affected	O
by	O
Bcr	B-Gene_or_gene_product
/	I-Gene_or_gene_product
Abl	I-Gene_or_gene_product
.	O

These	O
results	O
show	O
that	O
the	O
actin	B-Gene_or_gene_product
-	O
binding	O
domain	O
of	O
Abl	B-Gene_or_gene_product
enhances	O
leukemia	B-Cancer
development	O
but	O
does	O
not	O
appear	O
to	O
be	O
an	O
absolute	O
requirement	O
for	O
leukemogenesis	O
.	O

Building	O
the	O
vertebrate	O
vasculature	B-Anatomical_system
:	O
research	O
is	O
going	O
swimmingly	O
.	O

The	O
vertebrate	O
vasculature	B-Anatomical_system
develops	O
in	O
remarkably	O
similar	O
fashion	O
in	O
all	O
vertebrates	O
.	O

A	O
cohort	O
of	O
unspecified	O
mesodermal	B-Cell
cells	I-Cell
differentiates	O
into	O
primitive	O
endothelial	B-Cell
cells	I-Cell
,	O
which	O
migrate	O
to	O
and	O
occupy	O
positions	O
within	O
the	O
stereotypical	O
blueprint	O
of	O
the	O
primitive	O
vasculature	B-Anatomical_system
.	O

Once	O
in	O
position	O
,	O
these	O
cells	B-Cell
coalesce	O
and	O
form	O
cords	B-Tissue
,	O
which	O
lumenize	O
and	O
become	O
ensheathed	O
by	O
supporting	O
pericytes	B-Cell
and	O
smooth	B-Cell
muscle	I-Cell
cells	I-Cell
.	O

This	O
primitive	O
vascular	B-Anatomical_system
network	I-Anatomical_system
is	O
extensively	O
remodeled	O
in	O
some	O
places	O
,	O
and	O
expanded	O
by	O
sprouting	O
in	O
others	O
.	O

Various	O
studies	O
using	O
the	O
mouse	B-Organism
,	O
quail	B-Organism
/	O
chick	B-Organism
,	O
and	O
frog	B-Organism
have	O
uncovered	O
a	O
number	O
of	O
signals	O
that	O
guide	O
these	O
complex	O
processes	O
but	O
many	O
gaps	O
still	O
exist	O
in	O
our	O
understanding	O
of	O
the	O
mechanisms	O
by	O
which	O
the	O
embryonic	B-Anatomical_system
vasculature	I-Anatomical_system
is	O
built	O
.	O

Because	O
many	O
questions	O
will	O
require	O
in	O
vivo	O
studies	O
to	O
be	O
properly	O
addressed	O
,	O
the	O
zebrafish	B-Organism
,	O
with	O
its	O
unique	O
accessibility	O
to	O
analysis	O
by	O
combined	O
embryological	B-Developing_anatomical_structure
,	O
molecular	O
,	O
and	O
genetic	O
methods	O
,	O
should	O
prove	O
invaluable	O
in	O
identifying	O
new	O
molecules	O
involved	O
in	O
blood	B-Multi-tissue_structure
vessel	I-Multi-tissue_structure
development	O
and	O
integrating	O
pathways	O
that	O
influence	O
embryonic	B-Multi-tissue_structure
blood	I-Multi-tissue_structure
vessel	I-Multi-tissue_structure
formation	O
.	O

[	O
Interleukin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
18	I-Gene_or_gene_product
and	O
new	O
drugs	O
.	O
Protective	O
effect	O
against	O
tumor	B-Cancer
growth	O
and	O
infections	O
]	O
.	O

IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
18	I-Gene_or_gene_product
,	O
originally	O
identified	O
as	O
interferon	B-Gene_or_gene_product
-	I-Gene_or_gene_product
gamma	I-Gene_or_gene_product
inducing	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
IGIF	B-Gene_or_gene_product
)	O
,	O
is	O
related	O
to	O
the	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
family	O
in	O
terms	O
of	O
its	O
structure	O
as	O
well	O
as	O
its	O
processing	O
,	O
receptor	O
,	O
signal	O
transduction	O
pathway	O
and	O
pro	O
-	O
inflammatory	O
properties	O
.	O

IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
18	I-Gene_or_gene_product
is	O
also	O
functionally	O
related	O
to	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
12	I-Gene_or_gene_product
,	O
as	O
it	O
induces	O
the	O
production	O
of	O
Th1	B-Gene_or_gene_product
cytokines	O
and	O
participates	O
in	O
cell	B-Cell
-	O
mediated	O
immune	O
cytotoxicity	O
.	O

A	O
summary	O
is	O
made	O
of	O
recent	O
advances	O
in	O
the	O
understanding	O
of	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
18	I-Gene_or_gene_product
structure	O
,	O
processing	O
,	O
receptor	O
expression	O
and	O
immunoregulatory	O
functions	O
.	O

It	O
focuses	O
on	O
the	O
role	O
of	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
18	I-Gene_or_gene_product
modulation	O
in	O
tumours	B-Cancer
,	O
infections	O
and	O
autoimmune	O
and	O
inflammatory	O
diseases	O
.	O

A	O
small	O
peptide	O
derived	O
from	O
Flt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
functions	O
as	O
an	O
angiogenic	O
inhibitor	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
an	O
angiogenic	O
stimulator	O
which	O
functions	O
through	O
two	O
endothelial	B-Cell
specific	O
tyrosine	O
kinase	O
receptors	O
,	O
Flt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
Flk	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

In	O
this	O
work	O
,	O
we	O
show	O
that	O
an	O
11	O
-	O
amino	O
acid	O
peptide	O
derived	O
from	O
the	O
second	O
immunoglobulin	O
-	O
like	O
domain	O
of	O
Flt	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
functions	O
as	O
an	O
angiogenic	O
inhibitor	O
in	O
chick	B-Organism
chorioallantoic	B-Multi-tissue_structure
membrane	I-Multi-tissue_structure
and	O
inhibited	O
VEGF	B-Gene_or_gene_product
-	O
induced	O
vascular	B-Multi-tissue_structure
permeability	O
in	O
Miles	O
'	O
assay	O
without	O
binding	O
to	O
VEGF	B-Gene_or_gene_product
directly	O
.	O

Circular	O
dichroism	O
and	O
nuclear	O
magnetic	O
resonance	O
analyses	O
indicate	O
that	O
this	O
peptide	O
forms	O
a	O
stable	O
extended	O
structure	O
in	O
solution	O
,	O
presumably	O
beta	O
-	O
sheet	O
structure	O
and	O
is	O
most	O
likely	O
existing	O
as	O
a	O
dimer	O
.	O

Our	O
results	O
suggest	O
that	O
this	O
small	O
peptide	O
functions	O
as	O
an	O
angiogenic	O
inhibitor	O
by	O
inhibiting	O
VEGF	B-Gene_or_gene_product
function	O
through	O
a	O
non	O
-	O
VEGF	B-Gene_or_gene_product
binding	O
mechanism	O
.	O

Coexpression	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
p53	B-Gene_or_gene_product
protein	O
in	O
squamous	B-Cancer
cell	I-Cancer
carcinoma	I-Cancer
of	O
the	O
esophagus	B-Organ
.	O

OBJECTIVE	O
:	O
p53	B-Gene_or_gene_product
plays	O
a	O
role	O
in	O
tumor	B-Cancer
angiogenesis	O
,	O
and	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
plays	O
a	O
key	O
role	O
in	O
tumor	B-Cancer
angiogenesis	O
.	O

The	O
aim	O
of	O
the	O
present	O
study	O
was	O
to	O
clarify	O
how	O
expression	O
of	O
p53	B-Gene_or_gene_product
protein	O
participates	O
in	O
angiogenesis	O
,	O
and	O
whether	O
the	O
coexpression	O
of	O
VEGF	B-Gene_or_gene_product
and	O
p53	B-Gene_or_gene_product
protein	O
has	O
a	O
significance	O
for	O
angiogenesis	O
and	O
the	O
clinicopathological	O
features	O
in	O
esophageal	B-Cancer
squamous	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
(	O
SCC	B-Cancer
)	O
.	O

METHODS	O
:	O
Tissues	B-Tissue
samples	I-Tissue
were	O
taken	O
from	O
60	O
patients	B-Organism
with	O
esophageal	B-Cancer
SCC	I-Cancer
after	O
surgery	O
.	O

The	O
expression	O
of	O
VEGF	B-Gene_or_gene_product
and	O
p53	B-Gene_or_gene_product
protein	O
in	O
these	O
SCC	B-Cancer
was	O
examined	O
immunohistochemically	O
.	O

Microvessel	B-Tissue
density	O
(	O
MVD	O
)	O
was	O
determined	O
by	O
counting	O
microvessels	B-Tissue
in	O
tumor	B-Multi-tissue_structure
sections	I-Multi-tissue_structure
stained	O
for	O
Factor	B-Gene_or_gene_product
VIII	I-Gene_or_gene_product
-	I-Gene_or_gene_product
related	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
.	O

Ki	B-Gene_or_gene_product
-	I-Gene_or_gene_product
67	I-Gene_or_gene_product
labeling	O
index	O
(	O
LI	O
)	O
was	O
calculated	O
,	O
based	O
on	O
Ki	B-Gene_or_gene_product
-	I-Gene_or_gene_product
67	I-Gene_or_gene_product
antigen	O
immunostaining	O
,	O
as	O
a	O
proliferative	O
marker	O
.	O

Apoptotic	O
index	O
(	O
AI	O
)	O
was	O
calculated	O
,	O
based	O
on	O
the	O
terminal	B-Gene_or_gene_product
deoxynucleotidyl	I-Gene_or_gene_product
transferase	I-Gene_or_gene_product
-	O
mediated	O
deoxyuridine	B-Simple_chemical
triphosphate	I-Simple_chemical
biotin	I-Simple_chemical
nick	O
end	O
labeling	O
,	O
to	O
evaluate	O
apoptosis	O
.	O

RESULTS	O
:	O
VEGF	B-Gene_or_gene_product
expression	O
was	O
observed	O
in	O
58	O
.	O
3	O
%	O
,	O
and	O
p53	B-Gene_or_gene_product
protein	O
expression	O
was	O
observed	O
in	O
61	O
.	O
7	O
%	O
of	O
the	O
60	O
patients	B-Organism
.	O

VEGF	B-Gene_or_gene_product
and	O
p53	B-Gene_or_gene_product
protein	O
were	O
significantly	O
coexpressed	O
in	O
26	O
(	O
43	O
.	O
4	O
%	O
)	O
.	O

Histological	O
venous	B-Multi-tissue_structure
invasion	O
(	O
p	O
less	O
than	O
0	O
.	O
01	O
)	O
and	O
distant	O
metastasis	O
(	O
p	O
less	O
than	O
0	O
.	O
05	O
)	O
were	O
significantly	O
correlated	O
with	O
p53	B-Gene_or_gene_product
protein	O
expression	O
.	O

The	O
two	O
parameters	O
were	O
more	O
frequently	O
observed	O
in	O
the	O
SCC	B-Cancer
with	O
VEGF	B-Gene_or_gene_product
/	O
p53	B-Gene_or_gene_product
coexpression	O
than	O
in	O
those	O
without	O
the	O
coexpression	O
.	O

The	O
MVD	O
and	O
Ki	B-Gene_or_gene_product
-	I-Gene_or_gene_product
67	I-Gene_or_gene_product
LI	O
were	O
significantly	O
higher	O
(	O
p	O
less	O
than	O
0	O
.	O
01	O
and	O
p	O
less	O
than	O
0	O
.	O
001	O
)	O
,	O
and	O
the	O
AI	O
was	O
significantly	O
lower	O
(	O
p	O
less	O
than	O
0	O
.	O
001	O
)	O
in	O
the	O
SCC	B-Cancer
with	O
p53	B-Gene_or_gene_product
protein	O
expression	O
than	O
in	O
the	O
SCC	B-Cancer
without	O
it	O
.	O

The	O
MVD	O
and	O
Ki	B-Gene_or_gene_product
-	I-Gene_or_gene_product
67	I-Gene_or_gene_product
LI	O
were	O
higher	O
,	O
and	O
the	O
AI	O
was	O
lower	O
in	O
the	O
SCC	B-Cancer
with	O
VEGF	B-Gene_or_gene_product
/	O
p53	B-Gene_or_gene_product
coexpression	O
than	O
in	O
those	O
without	O
the	O
coexpression	O
.	O

The	O
5	O
-	O
yr	O
survival	O
rate	O
in	O
patients	B-Organism
with	O
the	O
coexpression	O
was	O
poorer	O
than	O
in	O
the	O
other	O
patients	B-Organism
.	O

CONCLUSION	O
:	O
These	O
results	O
suggest	O
that	O
mutant	O
p53	B-Gene_or_gene_product
expression	O
is	O
associated	O
with	O
angiogenesis	O
and	O
distant	O
metastasis	O
in	O
esophageal	B-Cancer
SCC	I-Cancer
,	O
and	O
that	O
the	O
coexpression	O
of	O
p53	B-Gene_or_gene_product
and	O
VEGF	B-Gene_or_gene_product
may	O
play	O
an	O
important	O
role	O
in	O
angiogenesis	O
,	O
and	O
have	O
important	O
clinical	O
significance	O
.	O

[	O
mRNA	O
expression	O
of	O
metastasis	O
-	O
suppressor	O
gene	O
nm23	B-Gene_or_gene_product
in	O
carcinoma	B-Cancer
of	O
buccal	B-Multi-tissue_structure
mucosa	I-Multi-tissue_structure
.	O
II	O
.	O
Quantitative	O
reverse	O
transcription	O
PCR	O
amplification	O
]	O
.	O

The	O
nm23	B-Gene_or_gene_product
gene	O
is	O
a	O
conspicuous	O
metastasis	O
-	O
suppressor	O
gene	O
.	O

The	O
authors	O
detected	O
both	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
and	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H2	I-Gene_or_gene_product
mRNA	O
levels	O
in	O
47	O
tissues	B-Tissue
samples	I-Tissue
of	O
patients	B-Organism
with	O
carcinoma	B-Cancer
of	I-Cancer
buccal	I-Cancer
mucosa	I-Cancer
(	O
CBM	B-Cancer
)	O
by	O
quantitative	O
reverse	O
transcription	O
PCR	O
amplification	O
.	O

The	O
results	O
showed	O
that	O
expression	O
levels	O
of	O
both	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
and	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H2	I-Gene_or_gene_product
mRNA	O
varied	O
in	O
normal	O
buccal	B-Multi-tissue_structure
mucosa	I-Multi-tissue_structure
,	O
leukoplakia	B-Pathological_formation
,	O
adjacent	O
nontumorous	B-Multi-tissue_structure
mucosa	I-Multi-tissue_structure
,	O
submandibular	B-Organ
gland	I-Organ
,	O
CBM	B-Cancer
and	O
lymph	B-Multi-tissue_structure
nodes	I-Multi-tissue_structure
with	O
or	O
without	O
metastasis	O
.	O

The	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
mRNA	O
expression	O
levels	O
in	O
CBM	B-Cancer
with	O
lymph	B-Multi-tissue_structure
nodes	I-Multi-tissue_structure
metastases	O
were	O
lower	O
than	O
those	O
in	O
CBM	B-Cancer
without	O
metastases	O
(	O
P	O
<	O
0	O
.	O
05	O
)	O
,	O
while	O
no	O
significance	O
of	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H2	I-Gene_or_gene_product
mRNA	O
expression	O
levels	O
was	O
found	O
between	O
CBM	B-Cancer
with	O
and	O
CBM	B-Cancer
without	O
metastasis	O
(	O
P	O
>	O
0	O
.	O
05	O
)	O
.	O

The	O
results	O
were	O
comparative	O
to	O
those	O
of	O
Northern	O
blotting	O
of	O
the	O
same	O
cases	O
.	O

The	O
authors	O
concluded	O
that	O
,	O
as	O
also	O
in	O
the	O
study	O
of	O
Northern	O
blotting	O
,	O
the	O
expression	O
of	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H1	I-Gene_or_gene_product
mRNA	O
significantly	O
correlated	O
inversely	O
with	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
in	O
CBM	B-Cancer
,	O
while	O
the	O
expression	O
of	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
H2	I-Gene_or_gene_product
mRNA	O
not	O
.	O

Q	O
-	O
RT	O
-	O
PCR	O
was	O
a	O
useful	O
method	O
to	O
detect	O
the	O
mRNA	O
levels	O
of	O
nm23	B-Gene_or_gene_product
gene	O
in	O
buccal	B-Cancer
carcinoma	I-Cancer
.	O

Neoplastic	O
transformation	O
and	O
tumorigenesis	O
associated	O
with	O
overexpression	O
of	O
phospholipase	B-Gene_or_gene_product
D	I-Gene_or_gene_product
isozymes	O
in	O
cultured	O
murine	B-Organism
fibroblasts	B-Cell
.	O

Phospholipase	B-Gene_or_gene_product
D	I-Gene_or_gene_product
(	O
PLD	B-Gene_or_gene_product
)	O
has	O
been	O
suggested	O
to	O
play	O
an	O
important	O
role	O
in	O
a	O
variety	O
of	O
cellular	B-Cell
functions	O
.	O

PLD	B-Gene_or_gene_product
activity	O
has	O
been	O
shown	O
to	O
be	O
significantly	O
elevated	O
in	O
many	O
tumours	B-Cancer
and	O
transformed	O
cells	B-Cell
,	O
suggesting	O
the	O
possibility	O
that	O
PLD	B-Gene_or_gene_product
might	O
be	O
involved	O
in	O
tumorigenesis	O
.	O

In	O
this	O
study	O
,	O
we	O
have	O
established	O
stable	O
cell	B-Cell
lines	I-Cell
overexpressing	O
PLD1	B-Gene_or_gene_product
and	O
PLD2	B-Gene_or_gene_product
from	O
fibroblast	B-Cell
cells	I-Cell
.	O

These	O
cells	B-Cell
,	O
but	O
not	O
control	O
cells	B-Cell
,	O
showed	O
altered	O
growth	O
properties	O
and	O
anchorage	O
-	O
independent	O
growth	O
in	O
soft	O
agar	O
.	O

Both	O
PLD1	B-Gene_or_gene_product
and	O
PLD2	B-Gene_or_gene_product
also	O
induced	O
an	O
up	O
-	O
regulation	O
of	O
the	O
activity	O
of	O
matrix	B-Gene_or_gene_product
metalloprotease	I-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
as	O
detected	O
by	O
zymograms	O
.	O

Furthermore	O
,	O
both	O
PLD1	B-Gene_or_gene_product
and	O
PLD2	B-Gene_or_gene_product
transformants	O
,	O
but	O
not	O
vector	B-Cell
-	I-Cell
transfectants	I-Cell
,	O
induced	O
undifferentiated	O
sarcoma	B-Cancer
when	O
transplanted	O
into	O
nude	B-Organism
mice	I-Organism
.	O

Both	O
PLD1	B-Gene_or_gene_product
-	O
and	O
PLD2	B-Gene_or_gene_product
-	O
mediated	O
cell	B-Cell
cycle	O
distributions	O
in	O
stable	O
cell	B-Cell
lines	I-Cell
revealed	O
an	O
increased	O
fraction	O
of	O
cells	B-Cell
in	O
the	O
S	O
phase	O
compared	O
with	O
control	O
cells	B-Cell
.	O

Interestingly	O
,	O
the	O
level	O
of	O
cyclin	B-Gene_or_gene_product
D3	I-Gene_or_gene_product
protein	O
,	O
known	O
as	O
an	O
activator	O
of	O
G	O
(	O
1	O
)	O
to	O
S	O
phase	O
transition	O
in	O
the	O
cell	B-Cell
cycle	O
,	O
was	O
aberrantly	O
high	O
in	O
cells	B-Cell
overexpressing	O
PLD1	B-Gene_or_gene_product
and	O
PLD2	B-Gene_or_gene_product
compared	O
with	O
control	O
cells	B-Cell
.	O

These	O
results	O
suggest	O
that	O
overexpression	O
of	O
PLD	B-Gene_or_gene_product
isozymes	O
may	O
play	O
an	O
important	O
role	O
in	O
neoplastic	O
transformation	O
.	O

Troponin	B-Gene_or_gene_product
I	I-Gene_or_gene_product
inhibits	O
capillary	B-Cell
endothelial	I-Cell
cell	I-Cell
proliferation	O
by	O
interaction	O
with	O
the	O
cell	B-Cell
'	O
s	O
bFGF	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
.	O

Troponin	B-Gene_or_gene_product
I	I-Gene_or_gene_product
(	O
TnI	B-Gene_or_gene_product
)	O
is	O
a	O
novel	O
cartilage	O
-	O
derived	O
angiogenesis	O
inhibitor	O
,	O
first	O
demonstrated	O
by	O
Moses	O
et	O
al	O
.	O

(	O
1999	O
,	O
Proc	O
.	O
Natl	O
.	O
Acad	O
.	O
Sci	O
.	O
USA	O
2645	O
-	O
2650	O
)	O
to	O
inhibit	O
endothelial	B-Cell
cell	I-Cell
proliferation	O
and	O
angiogenesis	O
,	O
both	O
in	O
vivo	O
and	O
in	O
vitro	O
,	O
and	O
to	O
inhibit	O
metastasis	O
of	O
a	O
wide	O
variety	O
of	O
tumors	B-Cancer
in	O
vivo	O
.	O

Despite	O
convincing	O
evidence	O
of	O
its	O
efficacy	O
,	O
little	O
is	O
known	O
about	O
the	O
mechanism	O
of	O
action	O
of	O
TnI	B-Gene_or_gene_product
as	O
an	O
anti	O
-	O
proliferative	O
and	O
anti	O
-	O
angiogenic	O
agent	O
.	O

In	O
the	O
current	O
article	O
we	O
demonstrate	O
that	O
TnI	B-Gene_or_gene_product
inhibits	O
both	O
bFGF	B-Gene_or_gene_product
-	O
stimulated	O
and	O
basal	O
levels	O
of	O
endothelial	B-Cell
cell	I-Cell
proliferation	O
,	O
and	O
we	O
hypothesize	O
that	O
this	O
inhibition	O
is	O
occurring	O
,	O
at	O
least	O
in	O
part	O
,	O
via	O
an	O
interaction	O
of	O
TnI	B-Gene_or_gene_product
with	O
the	O
cell	B-Cellular_component
-	I-Cellular_component
surface	I-Cellular_component
bFGF	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
on	O
capillary	B-Cell
endothelial	I-Cell
cells	I-Cell
.	O

We	O
further	O
support	O
this	O
hypothesis	O
by	O
providing	O
the	O
first	O
evidence	O
that	O
TnI	B-Gene_or_gene_product
can	O
act	O
on	O
nonendothelial	B-Cell
as	O
well	O
as	O
endothelial	B-Cell
cells	I-Cell
and	O
by	O
demonstrating	O
that	O
this	O
inhibitory	O
action	O
is	O
specific	O
for	O
the	O
bFGF	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
on	O
the	O
target	O
cells	B-Cell
.	O

Preliminary	O
data	O
suggest	O
that	O
TnI	B-Gene_or_gene_product
may	O
be	O
competing	O
with	O
bFGF	B-Gene_or_gene_product
for	O
interaction	O
with	O
the	O
bFGF	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
on	O
responsive	O
cells	B-Cell
.	O

RhoA	B-Gene_or_gene_product
activation	O
promotes	O
transformation	O
and	O
loss	O
of	O
thyroid	B-Cell
cell	I-Cell
differentiation	O
interfering	O
with	O
thyroid	B-Gene_or_gene_product
transcription	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
activity	O
.	O

Highly	O
specialized	O
cells	B-Cell
,	O
the	O
thyrocytes	B-Cell
,	O
express	O
a	O
thyroid	B-Organ
-	O
specific	O
set	O
of	O
genes	O
for	O
thyroglobulin	B-Gene_or_gene_product
(	O
Tg	B-Gene_or_gene_product
)	O
,	O
thyroperoxidase	B-Gene_or_gene_product
,	O
and	O
the	O
transcription	O
factors	O
TTF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
TTF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
and	O
Pax	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
.	O

The	O
implication	O
of	O
the	O
small	O
GTPase	O
RhoA	B-Gene_or_gene_product
in	O
TSH	B-Gene_or_gene_product
-	O
mediated	O
proliferation	O
of	O
FRTL	B-Cell
-	I-Cell
5	I-Cell
rat	I-Cell
thyroid	I-Cell
cells	I-Cell
has	O
been	O
previously	O
demonstrated	O
.	O

To	O
further	O
analyze	O
RhoA	B-Gene_or_gene_product
function	O
in	O
thyroid	B-Cell
cell	I-Cell
proliferation	O
and	O
differentiation	O
patterns	O
,	O
we	O
combined	O
transient	O
and	O
stable	O
transfection	O
assays	O
to	O
express	O
different	O
mutant	O
RhoA	B-Gene_or_gene_product
forms	O
in	O
FRTL	B-Cell
-	I-Cell
5	I-Cell
cells	I-Cell
.	O

Constitutively	O
active	O
RhoA	B-Gene_or_gene_product
(	O
FRTL	O
-	O
5	O
-	O
RhoA	B-Gene_or_gene_product
QL	O
cells	O
)	O
exhibited	O
a	O
fibroblast	B-Cell
-	O
like	O
phenotype	O
with	O
organized	O
actin	B-Gene_or_gene_product
fibers	B-Cellular_component
,	O
whereas	O
cells	B-Cell
expressing	O
the	O
RhoA	B-Gene_or_gene_product
negative	O
dominant	O
phenotype	O
(	O
FRTL	B-Cell
-	I-Cell
5	I-Cell
-	I-Cell
RhoA	I-Cell
N19	I-Cell
cells	I-Cell
)	O
present	O
a	O
rounded	O
morphology	O
and	O
lose	O
normal	O
cytoskeletal	B-Cellular_component
architecture	O
.	O

In	O
addition	O
,	O
expression	O
of	O
the	O
constitutively	O
active	O
form	O
of	O
RhoA	B-Gene_or_gene_product
results	O
in	O
TSH	B-Gene_or_gene_product
-	O
independent	O
proliferation	O
and	O
anchorage	O
-	O
independent	O
growth	O
and	O
induces	O
tumors	B-Cancer
when	O
inoculated	O
in	O
nude	B-Organism
mice	I-Organism
.	O

Interestingly	O
,	O
FRTL	O
-	O
5	O
-	O
RhoA	B-Gene_or_gene_product
QL	O
cells	O
express	O
less	O
Tg	B-Gene_or_gene_product
and	O
TTF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
than	O
wild	O
-	O
type	O
FRTL	B-Cell
-	I-Cell
5	I-Cell
(	O
FRTL	B-Cell
-	I-Cell
5	I-Cell
-	O
vector	O
)	O
or	O
FRTL	O
-	O
5	O
-	O
RhoA	B-Gene_or_gene_product
N19	O
,	O
suggesting	O
a	O
loss	O
at	O
the	O
differentiation	O
stage	O
.	O

This	O
effect	O
is	O
mediated	O
,	O
at	O
least	O
in	O
part	O
,	O
by	O
a	O
decrease	O
in	O
TTF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
activity	O
,	O
since	O
transient	O
or	O
stable	O
expression	O
of	O
RhoA	B-Gene_or_gene_product
QL	O
results	O
in	O
a	O
reduction	O
in	O
the	O
activity	O
of	O
the	O
wild	O
-	O
type	O
Tg	B-Gene_or_gene_product
promoter	O
as	O
well	O
as	O
an	O
artificial	O
promoter	O
the	O
activation	O
of	O
which	O
depends	O
exclusively	O
on	O
TTF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

The	O
similarity	O
between	O
RhoA	B-Gene_or_gene_product
effects	O
and	O
thyroid	B-Cell
transformation	O
by	O
Ras	B-Gene_or_gene_product
suggests	O
that	O
RhoA	B-Gene_or_gene_product
may	O
act	O
as	O
a	O
downstream	O
effector	O
of	O
Ras	B-Gene_or_gene_product
;	O
in	O
fact	O
,	O
the	O
dominant	O
negative	O
RhoA	B-Gene_or_gene_product
N19	O
abolished	O
the	O
down	O
-	O
regulatory	O
effect	O
of	O
Ras	B-Gene_or_gene_product
V12	O
over	O
the	O
Tg	B-Gene_or_gene_product
promoter	O
.	O

Taken	O
together	O
,	O
these	O
results	O
show	O
for	O
the	O
first	O
time	O
that	O
active	O
RhoA	B-Gene_or_gene_product
is	O
able	O
to	O
transform	O
FRTL	B-Cell
-	I-Cell
5	I-Cell
cells	I-Cell
and	O
that	O
this	O
effect	O
is	O
coupled	O
to	O
a	O
loss	O
of	O
thyroid	B-Cell
differentiation	O
due	O
to	O
impaired	O
TTF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
activity	O
.	O

Clinical	O
significance	O
of	O
plasma	B-Organism_substance
endostatin	B-Gene_or_gene_product
in	O
acute	B-Cancer
myeloid	I-Cancer
leukemia	I-Cancer
/	O
myelodysplastic	O
syndrome	O
.	O

BACKGROUND	O
:	O
Endostatin	B-Gene_or_gene_product
,	O
a	O
C	O
-	O
terminal	O
fragment	O
of	O
collagen	B-Gene_or_gene_product
XVIII	I-Gene_or_gene_product
,	O
is	O
an	O
endogenous	O
angiogenesis	O
inhibitor	O
.	O

While	O
endostatin	B-Gene_or_gene_product
is	O
being	O
investigated	O
for	O
its	O
usefulness	O
in	O
treating	O
solid	B-Cancer
tumors	I-Cancer
,	O
its	O
significance	O
in	O
hematologic	B-Cancer
malignancies	I-Cancer
is	O
unknown	O
.	O

METHODS	O
:	O
The	O
authors	O
evaluated	O
plasma	B-Organism_substance
endostatin	B-Gene_or_gene_product
(	O
PE	B-Gene_or_gene_product
)	O
levels	O
using	O
an	O
enzyme	O
linked	O
immunoassay	O
in	O
71	O
patients	B-Organism
with	O
acute	B-Cancer
myeloid	I-Cancer
leukemia	I-Cancer
(	O
AML	B-Cancer
)	O
and	O
43	O
patients	B-Organism
with	O
myelodysplastic	O
syndrome	O
(	O
MDS	O
)	O
,	O
and	O
correlated	O
PE	B-Gene_or_gene_product
with	O
various	O
clinical	O
parameters	O
.	O

RESULTS	O
:	O
There	O
was	O
no	O
significant	O
difference	O
in	O
the	O
median	O
PE	B-Gene_or_gene_product
level	O
between	O
AML	B-Cancer
/	O
MDS	O
patients	B-Organism
and	O
the	O
normal	O
controls	O
.	O

Nevertheless	O
,	O
patients	B-Organism
who	O
achieved	O
complete	O
remission	O
(	O
CR	O
)	O
had	O
a	O
significantly	O
lower	O
median	O
PE	B-Gene_or_gene_product
level	O
compared	O
to	O
those	O
who	O
did	O
not	O
.	O

In	O
multivariate	O
analysis	O
,	O
PE	B-Gene_or_gene_product
was	O
found	O
to	O
be	O
a	O
significant	O
(	O
P	O
=	O
0	O
.	O
03	O
)	O
predictor	O
of	O
overall	O
survival	O
(	O
OS	O
)	O
with	O
adjustment	O
of	O
the	O
other	O
baseline	O
covariates	O
,	O
including	O
patient	B-Organism
age	O
,	O
history	O
of	O
antecedent	O
hematologic	O
disorders	O
,	O
and	O
the	O
use	O
of	O
protective	O
environments	O
.	O

The	O
prognostic	O
value	O
of	O
PE	B-Gene_or_gene_product
was	O
also	O
evaluated	O
by	O
dividing	O
MDS	O
/	O
AML	B-Cancer
patients	B-Organism
into	O
high	O
and	O
low	O
PE	B-Gene_or_gene_product
groups	O
using	O
the	O
median	O
PE	B-Gene_or_gene_product
level	O
of	O
normal	O
controls	O
as	O
the	O
cut	O
-	O
off	O
.	O

The	O
authors	O
found	O
that	O
patients	B-Organism
in	O
the	O
high	O
PE	B-Gene_or_gene_product
group	O
survived	O
for	O
a	O
significantly	O
shorter	O
time	O
than	O
those	O
patients	B-Organism
in	O
the	O
low	O
PE	B-Gene_or_gene_product
group	O
.	O

CONCLUSIONS	O
:	O
PE	B-Gene_or_gene_product
is	O
a	O
useful	O
prognostic	O
predictor	O
of	O
CR	O
and	O
OS	O
for	O
AML	B-Cancer
/	O
MDS	O
patients	B-Organism
.	O

The	O
mechanism	O
underlying	O
the	O
association	O
between	O
high	O
PE	B-Gene_or_gene_product
and	O
poor	O
clinical	O
outcome	O
is	O
unclear	O
,	O
although	O
it	O
may	O
be	O
related	O
to	O
the	O
possible	O
PE	B-Gene_or_gene_product
reflection	O
of	O
tumor	B-Cancer
burden	O
.	O

Modulation	O
of	O
angiogenesis	O
and	O
progelatinase	B-Gene_or_gene_product
a	I-Gene_or_gene_product
by	O
thrombin	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
mimetics	O
and	O
antagonists	O
.	O

The	O
angiogenic	O
action	O
of	O
thrombin	B-Gene_or_gene_product
has	O
been	O
shown	O
to	O
be	O
mediated	O
by	O
activation	O
of	O
the	O
thrombin	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
.	O

In	O
this	O
report	O
we	O
studied	O
the	O
effects	O
of	O
SFLLR	B-Gene_or_gene_product
,	O
an	O
agonist	O
of	O
the	O
activated	O
thrombin	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
and	O
thrombin	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
peptide	O
and	O
non	O
peptide	O
antagonists	O
on	O
angiogenesis	O
in	O
the	O
chick	B-Organism
chorioallantoic	B-Multi-tissue_structure
membrane	I-Multi-tissue_structure
(	O
CAM	B-Multi-tissue_structure
)	O
system	O
.	O

As	O
antagonists	O
were	O
used	O
the	O
tripeptide	B-Gene_or_gene_product
FPR	I-Gene_or_gene_product
and	O
non	O
-	O
peptide	O
1	B-Simple_chemical
,	I-Simple_chemical
4	I-Simple_chemical
-	I-Simple_chemical
disubstituted	I-Simple_chemical
piperazine	I-Simple_chemical
derivatives	I-Simple_chemical
.	O

The	O
pentapeptide	B-Gene_or_gene_product
SFLLR	I-Gene_or_gene_product
,	O
like	O
thrombin	B-Gene_or_gene_product
,	O
caused	O
a	O
marked	O
stimulation	O
of	O
angiogenesis	O
in	O
the	O
CAM	B-Multi-tissue_structure
.	O

FPR	B-Gene_or_gene_product
and	O
the	O
piperazine	B-Simple_chemical
derivatives	I-Simple_chemical
caused	O
suppression	O
of	O
angiogenesis	O
and	O
in	O
combination	O
with	O
thrombin	B-Gene_or_gene_product
antagonized	O
its	O
angiogenic	O
effect	O
.	O

Thrombin	B-Gene_or_gene_product
and	O
SFLLR	B-Gene_or_gene_product
activated	O
progelatinase	B-Gene_or_gene_product
A	I-Gene_or_gene_product
(	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
)	O
in	O
the	O
culture	O
medium	O
of	O
human	B-Cell
umbilical	I-Cell
cord	I-Cell
endothelial	I-Cell
cells	I-Cell
(	O
HUVECs	B-Cell
)	O
.	O

MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
is	O
involved	O
in	O
the	O
early	O
steps	O
of	O
angiogenesis	O
leading	O
to	O
local	O
dissolution	O
of	O
basement	B-Cellular_component
membrane	I-Cellular_component
collagen	B-Gene_or_gene_product
and	O
migration	O
of	O
the	O
activated	O
endothelial	B-Cell
cells	I-Cell
.	O

FPR	B-Gene_or_gene_product
and	O
the	O
piperazine	B-Simple_chemical
derivatives	I-Simple_chemical
inhibited	O
the	O
activation	O
of	O
this	O
enzyme	O
.	O

They	O
also	O
antagonised	O
the	O
effects	O
of	O
both	O
thrombin	B-Gene_or_gene_product
and	O
SFLLR	B-Gene_or_gene_product
on	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
activation	O
.	O

These	O
results	O
suggest	O
that	O
non	O
-	O
thrombogenic	O
agonists	O
or	O
antagonists	O
of	O
the	O
activated	O
thrombin	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
can	O
be	O
used	O
as	O
modulators	O
of	O
angiogenesis	O
.	O

Abnormal	O
expression	O
of	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
and	O
beta	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
may	O
be	O
a	O
molecular	O
marker	O
of	O
submucosal	B-Multi-tissue_structure
invasion	O
and	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
in	O
early	O
gastric	B-Cancer
cancer	I-Cancer
.	O

BACKGROUND	O
:	O
Impaired	O
expression	O
of	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
and	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
and	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
is	O
frequently	O
observed	O
in	O
several	O
human	B-Organism
cancers	B-Cancer
.	O

The	O
aim	O
of	O
this	O
study	O
was	O
to	O
examine	O
immunohistochemical	O
expression	O
of	O
these	O
adhesion	O
molecules	O
,	O
focusing	O
on	O
early	O
gastric	B-Cancer
carcinomas	I-Cancer
,	O
and	O
to	O
investigate	O
differences	O
between	O
differentiated	O
and	O
undifferentiated	O
gastric	B-Cancer
cancer	I-Cancer
at	O
the	O
early	O
phase	O
of	O
carcinogenesis	O
.	O

METHODS	O
:	O
Immunohistochemical	O
staining	O
of	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
and	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
and	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
was	O
performed	O
using	O
specimens	B-Multi-tissue_structure
from	O
143	O
patients	B-Organism
with	O
early	O
gastric	B-Cancer
cancer	I-Cancer
.	O

RESULTS	O
:	O
Abnormal	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
and	O
beta	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
staining	O
correlated	O
with	O
depth	O
of	O
tumour	B-Cancer
invasion	O
in	O
differentiated	O
-	O
type	O
tumours	B-Cancer
.	O

In	O
contrast	O
,	O
abnormal	O
staining	O
was	O
frequently	O
found	O
even	O
in	O
intramucosal	B-Cancer
carcinoma	I-Cancer
of	O
undifferentiated	O
-	O
type	O
tumours	B-Cancer
,	O
suggesting	O
an	O
apparent	O
difference	O
in	O
the	O
onset	O
of	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
-	O
catenin	B-Gene_or_gene_product
complex	O
abnormality	O
between	O
the	O
two	O
cancer	B-Cancer
types	O
.	O

Absent	O
staining	O
of	O
beta	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
was	O
associated	O
with	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
.	O

Multivariate	O
analysis	O
revealed	O
abnormal	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
expression	O
as	O
an	O
independent	O
factor	O
that	O
correlated	O
with	O
submucosal	B-Multi-tissue_structure
invasion	O
in	O
early	O
gastric	B-Cancer
cancer	I-Cancer
.	O

CONCLUSION	O
:	O
Abnormal	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
expression	O
is	O
a	O
possible	O
marker	O
of	O
submucosal	B-Multi-tissue_structure
invasion	O
in	O
differentiated	O
-	O
type	O
early	O
gastric	B-Cancer
cancer	I-Cancer
and	O
absent	O
beta	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
staining	O
could	O
be	O
used	O
as	O
a	O
predictor	O
of	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
in	O
both	O
types	O
.	O

Dominant	O
negative	O
interference	O
of	O
transcription	O
factor	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
causes	O
inhibition	O
of	O
ErbB	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
expression	O
and	O
suppresses	O
malignant	B-Cell
cell	I-Cell
growth	O
.	O

ErbB	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
(	O
HER3	B-Gene_or_gene_product
)	O
is	O
a	O
member	O
of	O
the	O
epidermal	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
family	O
.	O

Increasing	O
evidence	O
suggests	O
that	O
elevated	O
expression	O
of	O
ErbB	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
is	O
important	O
for	O
malignancy	O
.	O

In	O
this	O
study	O
,	O
we	O
found	O
that	O
elevated	O
levels	O
of	O
ErbB	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
expression	O
did	O
not	O
occur	O
in	O
the	O
absence	O
of	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2gamma	I-Gene_or_gene_product
in	O
a	O
panel	O
of	O
human	B-Organism
mammary	B-Cell
epithelial	I-Cell
and	O
fibroblasts	B-Cell
cell	I-Cell
lines	I-Cell
.	O

In	O
contrast	O
,	O
there	O
was	O
no	O
association	O
between	O
the	O
expression	O
of	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2alpha	I-Gene_or_gene_product
or	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2beta	I-Gene_or_gene_product
and	O
the	O
level	O
of	O
ErbB	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
,	O
or	O
between	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2alpha	I-Gene_or_gene_product
and	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2gamma	I-Gene_or_gene_product
double	O
positivity	O
and	O
ErbB	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
expression	O
.	O

In	O
co	O
-	O
transfection	O
experiments	O
,	O
exogenous	O
expression	O
of	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2gamma	I-Gene_or_gene_product
robustly	O
activated	O
ErbB	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
promoter	O
activity	O
.	O

Moreover	O
,	O
expression	O
of	O
a	O
dominant	O
negative	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
protein	O
,	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2delta	I-Gene_or_gene_product
(	O
deleted	O
residues	O
31	O
-	O
117	O
)	O
,	O
not	O
only	O
repressed	O
the	O
ErbB	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
promoter	O
activity	O
but	O
also	O
suppressed	O
endogenous	O
ErbB	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
transcription	O
in	O
the	O
ErbB	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
overexpressing	O
cell	B-Cell
line	I-Cell
MRC	I-Cell
-	I-Cell
5VA	I-Cell
.	O

Overexpression	O
of	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2A	I-Gene_or_gene_product
resulted	O
in	O
a	O
decreased	O
proliferation	O
rate	O
and	O
inhibitin	O
of	O
colony	B-Cell
formation	O
.	O

Taken	O
together	O
,	O
these	O
data	O
strongly	O
support	O
a	O
role	O
for	O
the	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
gene	O
family	O
,	O
in	O
particular	O
,	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2gamma	I-Gene_or_gene_product
,	O
in	O
the	O
control	O
of	O
ErbB	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
expression	O
.	O

Interference	O
with	O
the	O
function	O
of	O
transcription	O
factor	O
AP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
might	O
provide	O
a	O
potential	O
strategy	O
for	O
modulation	O
of	O
the	O
malignant	O
phenotype	O
.	O

VEGF165	B-Gene_or_gene_product
mediates	O
formation	O
of	O
complexes	O
containing	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
neuropilin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
that	O
enhance	O
VEGF165	B-Gene_or_gene_product
-	O
receptor	O
binding	O
.	O

Co	O
-	O
expression	O
of	O
NRP1	B-Gene_or_gene_product
and	O
(	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
)	O
KDR	B-Gene_or_gene_product
on	O
the	O
surface	B-Cellular_component
of	O
endothelial	B-Cell
cells	I-Cell
(	O
EC	B-Cell
)	O
enhances	O
VEGF165	B-Gene_or_gene_product
binding	O
to	O
KDR	B-Gene_or_gene_product
and	O
EC	B-Cell
chemotaxis	O
in	O
response	O
to	O
VEGF165	B-Gene_or_gene_product
.	O

Overexpression	O
of	O
NRP1	B-Gene_or_gene_product
by	O
prostate	B-Cell
tumor	I-Cell
cells	I-Cell
in	O
vivo	O
results	O
in	O
increased	O
tumor	B-Cancer
angiogenesis	O
and	O
growth	O
.	O

We	O
investigated	O
the	O
molecular	O
mechanisms	O
underlying	O
NRP1	B-Gene_or_gene_product
-	O
mediated	O
angiogenesis	O
by	O
analyzing	O
the	O
association	O
of	O
NRP1	B-Gene_or_gene_product
and	O
KDR	B-Gene_or_gene_product
.	O

An	O
intracellular	B-Immaterial_anatomical_entity
complex	O
containing	O
NRP1	B-Gene_or_gene_product
and	O
KDR	B-Gene_or_gene_product
was	O
immunoprecipitated	O
from	O
EC	B-Cell
by	O
anti	B-Gene_or_gene_product
-	I-Gene_or_gene_product
NRP1	I-Gene_or_gene_product
antibodies	I-Gene_or_gene_product
only	O
in	O
the	O
presence	O
of	O
VEGF165	B-Gene_or_gene_product
.	O

In	O
contrast	O
,	O
VEGF121	B-Gene_or_gene_product
,	O
which	O
does	O
not	O
bind	O
to	O
NRP1	B-Gene_or_gene_product
,	O
did	O
not	O
support	O
complex	O
formation	O
.	O

Complexes	O
containing	O
VEGF165	B-Gene_or_gene_product
,	O
NRP1	B-Gene_or_gene_product
,	O
and	O
KDR	B-Gene_or_gene_product
were	O
also	O
formed	O
in	O
an	O
intercellular	B-Immaterial_anatomical_entity
fashion	O
by	O
co	O
-	O
culture	O
of	O
EC	B-Cell
expressing	O
KDR	B-Gene_or_gene_product
only	O
,	O
with	O
cells	B-Cell
expressing	O
NRP1	B-Gene_or_gene_product
only	O
,	O
for	O
example	O
,	O
breast	B-Cell
carcinoma	I-Cell
cells	I-Cell
.	O

VEGF165	B-Gene_or_gene_product
also	O
mediated	O
the	O
binding	O
of	O
a	O
soluble	O
NRP1	B-Gene_or_gene_product
dimer	O
to	O
cells	B-Cell
expressing	O
KDR	B-Gene_or_gene_product
only	O
,	O
confirming	O
the	O
formation	O
of	O
such	O
complexes	O
.	O

Furthermore	O
,	O
the	O
formation	O
of	O
complexes	O
containing	O
KDR	B-Gene_or_gene_product
and	O
NRP1	B-Gene_or_gene_product
markedly	O
increased	O
125I	B-Gene_or_gene_product
-	I-Gene_or_gene_product
VEGF165	I-Gene_or_gene_product
binding	O
to	O
KDR	B-Gene_or_gene_product
.	O

Our	O
results	O
suggest	O
that	O
formation	O
of	O
a	O
ternary	O
complex	O
of	O
VEGF165	B-Gene_or_gene_product
,	O
KDR	B-Gene_or_gene_product
,	O
and	O
NRP1	B-Gene_or_gene_product
potentiates	O
VEGF165	B-Gene_or_gene_product
binding	O
to	O
KDR	B-Gene_or_gene_product
.	O

These	O
complexes	O
are	O
formed	O
on	O
the	O
surface	B-Cellular_component
of	O
EC	B-Cell
and	O
in	O
a	O
juxtacrine	O
manner	O
via	O
association	O
of	O
tumor	B-Cell
cell	I-Cell
NRP1	B-Gene_or_gene_product
and	O
EC	B-Cell
KDR	B-Gene_or_gene_product
.	O

[	O
Radiologic	O
-	O
pathologic	O
correlations	O
in	O
breast	B-Organism_subdivision
diseases	O
]	O

The	O
objective	O
of	O
this	O
article	O
is	O
to	O
explain	O
radiologic	O
patterns	O
of	O
benign	B-Pathological_formation
and	O
malignant	B-Pathological_formation
breast	I-Pathological_formation
lesions	I-Pathological_formation
(	O
masses	B-Pathological_formation
,	O
microcalcifications	B-Pathological_formation
)	O
based	O
on	O
histological	O
correlations	O
.	O

The	O
stromal	B-Tissue
fibrous	I-Tissue
reaction	O
associated	O
to	O
infiltrating	O
carcinomas	B-Cancer
is	O
responsible	O
of	O
focal	O
increased	O
density	O
,	O
and	O
architectural	O
distorsion	O
,	O
ultrasound	O
acoustic	O
shadowing	O
;	O
abnormal	O
neoangiogenesis	O
can	O
be	O
detected	O
by	O
Doppler	O
,	O
CT	O
or	O
MR	O
imaging	O
.	O

Invasive	O
carcinomas	B-Cancer
without	O
spiculated	B-Multi-tissue_structure
margins	I-Multi-tissue_structure
are	O
poorly	O
differentiated	O
tumors	B-Cancer
.	O

Mammographic	O
patterns	O
of	O
microcalcifications	B-Pathological_formation
depend	O
on	O
their	O
physiopathological	O
process	O
(	O
necrosis	O
,	O
secretion	O
)	O
,	O
and	O
the	O
shape	O
of	O
clusters	B-Pathological_formation
(	O
round	O
,	O
triangular	O
)	O
typifies	O
their	O
anatomical	O
site	O
of	O
origin	O
(	O
lobular	B-Multi-tissue_structure
,	O
ductal	B-Multi-tissue_structure
)	O
.	O

Less	O
frequent	O
lesions	B-Pathological_formation
(	O
invasive	O
lobular	B-Cancer
,	O
mucinous	B-Cancer
,	O
and	O
medullary	B-Cancer
carcinomas	I-Cancer
,	O
radial	B-Pathological_formation
scar	I-Pathological_formation
)	O
will	O
be	O
also	O
explained	O
based	O
on	O
radiopathological	O
correlations	O
.	O

Knowledge	O
of	O
radiopathological	O
correlations	O
in	O
breast	B-Organism_subdivision
diseases	O
helps	O
the	O
radiologists	O
to	O
analyze	O
and	O
characterize	O
breast	B-Pathological_formation
lesions	I-Pathological_formation
.	O

Pten	B-Gene_or_gene_product
signaling	O
in	O
gliomas	B-Cancer
.	O

In	O
1997	O
,	O
the	O
PTEN	B-Gene_or_gene_product
gene	O
(	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
was	O
identified	O
as	O
a	O
tumor	B-Cancer
suppressor	O
gene	O
on	O
the	O
long	O
arm	O
of	O
chromosome	B-Cellular_component
10	I-Cellular_component
.	O

Since	O
then	O
,	O
important	O
progress	O
has	O
been	O
made	O
with	O
respect	O
to	O
the	O
understanding	O
of	O
the	O
role	O
of	O
the	O
Pten	B-Gene_or_gene_product
protein	O
in	O
the	O
normal	O
development	O
of	O
the	O
brain	B-Organ
as	O
well	O
as	O
in	O
the	O
molecular	O
pathogenesis	O
of	O
human	B-Organism
gliomas	B-Cancer
.	O

This	O
review	O
summarizes	O
the	O
current	O
state	O
of	O
the	O
art	O
concerning	O
the	O
involvement	O
of	O
aberrant	O
Pten	B-Gene_or_gene_product
function	O
in	O
the	O
development	O
of	O
different	O
biologic	O
features	O
of	O
malignant	B-Cancer
gliomas	I-Cancer
,	O
such	O
as	O
loss	O
of	O
cell	B-Cell
-	O
cycle	O
control	O
and	O
uncontrolled	O
cell	B-Cell
proliferation	O
,	O
escape	O
from	O
apoptosis	O
,	O
brain	B-Organ
invasion	O
,	O
and	O
aberrant	O
neoangiogenesis	O
.	O

Most	O
of	O
the	O
tumor	B-Cancer
-	O
suppressive	O
properties	O
of	O
Pten	B-Gene_or_gene_product
are	O
dependent	O
on	O
its	O
lipid	B-Simple_chemical
phosphatase	O
activity	O
,	O
which	O
inhibits	O
the	O
phosphatidylinositol	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
'	I-Gene_or_gene_product
-	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
(	O
PI3K	B-Gene_or_gene_product
)	O
/	O
Akt	B-Gene_or_gene_product
signaling	O
pathway	O
through	O
dephosphorylation	O
of	O
phosphatidylinositol	B-Simple_chemical
-	I-Simple_chemical
(	I-Simple_chemical
3	I-Simple_chemical
,	I-Simple_chemical
4	I-Simple_chemical
,	I-Simple_chemical
5	I-Simple_chemical
)	I-Simple_chemical
-	I-Simple_chemical
triphosphate	I-Simple_chemical
.	O

The	O
additional	O
function	O
of	O
Pten	B-Gene_or_gene_product
as	O
a	O
dual	O
-	O
specificity	O
protein	O
phosphatase	O
may	O
also	O
play	O
a	O
role	O
in	O
glioma	B-Cancer
pathogenesis	O
.	O

Besides	O
the	O
wealth	O
of	O
data	O
elucidating	O
the	O
functional	O
roles	O
of	O
Pten	B-Gene_or_gene_product
,	O
recent	O
studies	O
suggest	O
a	O
diagnostic	O
significance	O
of	O
PTEN	B-Gene_or_gene_product
gene	O
alterations	O
as	O
a	O
molecular	O
marker	O
for	O
poor	O
prognosis	O
in	O
anaplastic	B-Cancer
astrocytomas	I-Cancer
and	O
anaplastic	B-Cancer
oligodendrogliomas	I-Cancer
.	O

Furthermore	O
,	O
the	O
possibility	O
of	O
selective	O
targeting	O
of	O
PTEN	B-Gene_or_gene_product
mutant	O
tumor	O
cells	O
by	O
specific	O
pharmacologic	O
inhibitors	O
of	O
members	O
of	O
the	O
Pten	B-Gene_or_gene_product
/	O
PI3K	B-Gene_or_gene_product
/	O
Akt	B-Gene_or_gene_product
pathway	O
opens	O
up	O
new	O
perspectives	O
for	O
a	O
targeted	O
molecular	O
therapy	O
of	O
malignant	B-Cancer
gliomas	I-Cancer
.	O

[	O
A	O
case	O
report	O
of	O
advanced	O
gastric	B-Cancer
cancer	I-Cancer
responding	O
to	O
TS	B-Simple_chemical
-	I-Simple_chemical
1	I-Simple_chemical
,	O
a	O
novel	O
oral	B-Organism_subdivision
fluorouracil	B-Simple_chemical
derivative	O
]	O
.	O

TS	B-Simple_chemical
-	I-Simple_chemical
1	I-Simple_chemical
is	O
a	O
new	O
,	O
oral	B-Organism_subdivision
anticancer	B-Cancer
agent	O
composed	O
of	O
two	O
modulators	O
,	O
gimeracil	B-Simple_chemical
(	O
CDHP	B-Simple_chemical
)	O
and	O
oteracil	B-Simple_chemical
potassium	I-Simple_chemical
(	O
Oxo	B-Simple_chemical
)	O
are	O
mixed	O
with	O
tegafur	B-Simple_chemical
in	O
a	O
ratio	O
of	O
1	O
:	O
0	O
.	O
4	O
:	O
1	O
.	O

We	O
report	O
one	O
case	O
of	O
advanced	O
gastric	B-Cancer
cancer	I-Cancer
with	O
lung	B-Organ
and	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastases	O
that	O
completely	O
responded	O
to	O
TS	B-Simple_chemical
-	I-Simple_chemical
1	I-Simple_chemical
.	O

A	O
71	O
-	O
year	O
-	O
old	O
woman	B-Organism
was	O
admitted	O
to	O
our	O
hospital	O
because	O
of	O
breathlessness	O
.	O

A	O
diagnosis	O
of	O
advanced	O
gastric	B-Cancer
cancer	I-Cancer
with	O
extensive	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastases	O
and	O
multiple	O
pulmonary	B-Organ
metastases	O
was	O
made	O
.	O

One	O
hundred	O
mg	O
/	O
body	O
/	O
day	O
of	O
TS	B-Simple_chemical
-	I-Simple_chemical
1	I-Simple_chemical
was	O
orally	O
administrated	O
for	O
4	O
weeks	O
.	O

A	O
partial	O
response	O
(	O
PR	O
)	O
was	O
obtained	O
after	O
the	O
first	O
course	O
with	O
regression	O
of	O
multiple	O
pulmonary	B-Organ
metastases	O
.	O

After	O
1	O
drug	O
-	O
free	O
week	O
,	O
the	O
second	O
course	O
was	O
administered	O
with	O
120	O
mg	O
/	O
body	O
/	O
day	O
of	O
TS	B-Simple_chemical
-	I-Simple_chemical
1	I-Simple_chemical
for	O
4	O
weeks	O
.	O

After	O
two	O
courses	O
,	O
the	O
primary	O
tumor	B-Cancer
was	O
reduced	O
to	O
an	O
ulcer	B-Pathological_formation
scar	I-Pathological_formation
with	O
pathological	O
confirmation	O
of	O
a	O
complete	O
disappearance	O
of	O
the	O
cancer	B-Tissue
tissue	I-Tissue
.	O

Moreover	O
,	O
computed	O
tomography	O
(	O
CT	O
)	O
showed	O
a	O
complete	O
regression	O
of	O
the	O
extensive	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
and	O
diffuse	O
lung	B-Organ
metastases	O
,	O
for	O
a	O
complete	O
response	O
(	O
CR	O
)	O
.	O

The	O
serum	B-Organism_substance
level	O
of	O
CEA	B-Gene_or_gene_product
was	O
reduced	O
from	O
172	O
.	O
7	O
ng	O
/	O
ml	O
to	O
8	O
.	O
1	O
ng	O
/	O
ml	O
after	O
TS	B-Simple_chemical
-	I-Simple_chemical
1	I-Simple_chemical
treatment	O
.	O

As	O
for	O
adverse	O
events	O
,	O
only	O
pigmentation	B-Pathological_formation
of	O
the	O
skin	B-Organ
and	O
Grade	O
2	O
oral	B-Pathological_formation
aphta	I-Pathological_formation
were	O
observed	O
.	O

Systemic	O
regulation	O
of	O
distraction	O
osteogenesis	O
:	O
a	O
cascade	O
of	O
biochemical	O
factors	O
.	O

This	O
study	O
investigates	O
the	O
systemic	O
biochemical	O
regulation	O
of	O
fracture	B-Pathological_formation
healing	O
in	O
distraction	O
osteogenesis	O
compared	O
with	O
rigid	O
osteotomy	O
in	O
a	O
prospective	O
in	O
vivo	O
study	O
in	O
humans	B-Organism
.	O

To	O
further	O
clarify	O
the	O
influence	O
of	O
mechanical	O
strain	O
on	O
the	O
regulation	O
of	O
bone	B-Organ
formation	O
,	O
bone	B-Organ
growth	O
factors	O
(	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	I-Gene_or_gene_product
,	O
IGF	B-Gene_or_gene_product
binding	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
[	I-Gene_or_gene_product
IGFBP	I-Gene_or_gene_product
]	I-Gene_or_gene_product
3	I-Gene_or_gene_product
,	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
beta1	I-Gene_or_gene_product
,	O
and	O
basic	B-Gene_or_gene_product
FGF	I-Gene_or_gene_product
[	O
bFGF	B-Gene_or_gene_product
]	O
)	O
,	O
bone	B-Cellular_component
matrix	I-Cellular_component
degrading	O
enzymes	O
(	O
matrix	B-Gene_or_gene_product
-	I-Gene_or_gene_product
metalloproteinases	I-Gene_or_gene_product
[	I-Gene_or_gene_product
MMPs	I-Gene_or_gene_product
]	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
2	B-Gene_or_gene_product
,	O
and	O
3	B-Gene_or_gene_product
)	O
,	O
human	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
hormone	I-Gene_or_gene_product
(	O
hGH	B-Gene_or_gene_product
)	O
,	O
and	O
bone	B-Organ
formation	O
markers	O
(	O
ALP	B-Gene_or_gene_product
,	O
bone	B-Gene_or_gene_product
-	I-Gene_or_gene_product
specific	I-Gene_or_gene_product
ALP	I-Gene_or_gene_product
[	O
BAP	B-Gene_or_gene_product
]	O
,	O
and	O
osteocalcin	B-Gene_or_gene_product
[	O
OC	B-Gene_or_gene_product
]	O
)	O
have	O
been	O
analyzed	O
in	O
serum	B-Organism_substance
samples	I-Organism_substance
from	O
10	O
patients	B-Organism
in	O
each	O
group	O
pre	O
-	O
and	O
postoperatively	O
.	O

In	O
the	O
distraction	O
group	O
,	O
a	O
significant	O
postoperative	O
increase	O
in	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
bFGF	B-Gene_or_gene_product
,	O
ALP	B-Gene_or_gene_product
,	O
and	O
BAP	B-Gene_or_gene_product
could	O
be	O
observed	O
during	O
the	O
lengthening	O
and	O
the	O
consolidation	O
period	O
when	O
compared	O
with	O
the	O
baseline	O
levels	O
.	O

Osteotomy	O
fracture	O
healing	O
without	O
the	O
traction	O
stimulus	O
failed	O
to	O
induce	O
a	O
corresponding	O
increase	O
in	O
these	O
factors	O
.	O

In	O
addition	O
,	O
comparison	O
of	O
both	O
groups	O
revealed	O
a	O
significantly	O
higher	O
increase	O
in	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta1	I-Gene_or_gene_product
,	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
I	I-Gene_or_gene_product
,	O
IGFBP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
,	O
and	O
hGH	B-Gene_or_gene_product
in	O
the	O
lengthening	O
group	O
during	O
the	O
distraction	O
period	O
,	O
indicating	O
key	O
regulatory	O
functions	O
in	O
mechanotransduction	O
.	O

The	O
time	O
courses	O
of	O
changes	O
in	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
bone	B-Organ
growth	O
factors	O
(	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta1	I-Gene_or_gene_product
and	O
bFGF	B-Gene_or_gene_product
)	O
,	O
and	O
hGH	B-Gene_or_gene_product
,	O
respectively	O
,	O
correlated	O
significantly	O
during	O
the	O
lengthening	O
phase	O
,	O
indicating	O
common	O
regulatory	O
pathways	O
for	O
these	O
factors	O
in	O
distraction	O
osteogenesis	O
.	O

Significant	O
correlation	O
between	O
the	O
osteoblastic	O
marker	O
BAP	B-Gene_or_gene_product
,	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta1	I-Gene_or_gene_product
,	O
and	O
bFGF	B-Gene_or_gene_product
suggests	O
strain	O
-	O
activated	O
osteoblastic	B-Cell
cells	I-Cell
as	O
a	O
major	O
source	O
of	O
systemically	O
increased	O
bone	B-Organ
growth	O
factors	O
during	O
callus	O
distraction	O
.	O

The	O
systemic	O
increase	O
in	O
bFGF	B-Gene_or_gene_product
and	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
might	O
reflect	O
an	O
increased	O
local	O
stimulation	O
of	O
angiogenesis	O
during	O
distraction	O
osteogenesis	O
.	O

Human	B-Organism
papillomavirus	I-Organism
oncoprotein	O
E6	B-Gene_or_gene_product
inactivates	O
the	O
transcriptional	O
coactivator	O
human	B-Organism
ADA3	B-Gene_or_gene_product
.	O

High	O
-	O
risk	O
human	B-Organism
papillomaviruses	I-Organism
(	O
HPVs	B-Organism
)	O
are	O
associated	O
with	O
carcinomas	B-Cancer
of	O
the	O
cervix	B-Multi-tissue_structure
and	O
other	O
genital	B-Cancer
tumors	I-Cancer
.	O

The	O
HPV	B-Organism
oncoprotein	O
E6	B-Gene_or_gene_product
is	O
essential	O
for	O
oncogenic	O
transformation	O
.	O

We	O
identify	O
here	O
hADA3	B-Gene_or_gene_product
,	O
human	B-Organism
homologue	O
of	O
the	O
yeast	B-Organism
transcriptional	O
coactivator	O
yADA3	B-Gene_or_gene_product
,	O
as	O
a	O
novel	O
E6	B-Gene_or_gene_product
-	O
interacting	O
protein	O
and	O
a	O
target	O
of	O
E6	B-Gene_or_gene_product
-	O
induced	O
degradation	O
.	O

hADA3	B-Gene_or_gene_product
binds	O
selectively	O
to	O
the	O
high	O
-	O
risk	O
HPV	B-Organism
E6	B-Gene_or_gene_product
proteins	O
and	O
only	O
to	O
immortalization	O
-	O
competent	O
E6	B-Gene_or_gene_product
mutants	O
.	O

hADA3	B-Gene_or_gene_product
functions	O
as	O
a	O
coactivator	O
for	O
p53	B-Gene_or_gene_product
-	O
mediated	O
transactivation	O
by	O
stabilizing	O
p53	B-Gene_or_gene_product
protein	O
.	O

Notably	O
,	O
three	O
immortalizing	O
E6	B-Gene_or_gene_product
mutants	O
that	O
do	O
not	O
induce	O
direct	O
p53	B-Gene_or_gene_product
degradation	O
but	O
do	O
interact	O
with	O
hADA3	B-Gene_or_gene_product
induced	O
the	O
abrogation	O
of	O
p53	B-Gene_or_gene_product
-	O
mediated	O
transactivation	O
and	O
G	O
(	O
1	O
)	O
cell	B-Cell
cycle	O
arrest	O
after	O
DNA	B-Cellular_component
damage	O
,	O
comparable	O
to	O
wild	O
-	O
type	O
E6	B-Gene_or_gene_product
.	O

These	O
findings	O
reveal	O
a	O
novel	O
strategy	O
of	O
HPV	B-Organism
E6	B-Gene_or_gene_product
-	O
induced	O
loss	O
of	O
p53	B-Gene_or_gene_product
function	O
that	O
is	O
independent	O
of	O
direct	O
p53	B-Gene_or_gene_product
degradation	O
.	O

Given	O
the	O
likely	O
role	O
of	O
the	O
evolutionarily	O
conserved	O
hADA3	B-Gene_or_gene_product
in	O
multiple	O
coactivator	O
complexes	O
,	O
inactivation	O
of	O
its	O
function	O
may	O
allow	O
E6	B-Gene_or_gene_product
to	O
perturb	O
numerous	O
cellular	B-Cell
pathways	O
during	O
HPV	B-Organism
oncogenesis	O
.	O

Immunohistochemical	O
staining	O
for	O
thyroid	B-Gene_or_gene_product
transcription	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
:	O
a	O
helpful	O
aid	O
in	O
discerning	O
primary	O
site	O
of	O
tumor	B-Cancer
origin	O
in	O
patients	B-Organism
with	O
brain	B-Organ
metastases	O
.	O

Metastatic	O
carcinoma	B-Cancer
of	O
unknown	O
primary	O
origin	O
is	O
a	O
perplexing	O
but	O
common	O
problem	O
,	O
accounting	O
for	O
up	O
to	O
10	O
%	O
to	O
15	O
%	O
of	O
all	O
solid	B-Cancer
tumors	I-Cancer
at	O
presentation	O
.	O

Many	O
of	O
these	O
metastases	O
presumably	O
arise	O
from	O
primary	O
lung	B-Cancer
carcinomas	I-Cancer
,	O
but	O
the	O
morphologic	O
features	O
and	O
immunohistochemical	O
profile	O
of	O
lung	B-Cancer
cancer	I-Cancer
is	O
often	O
too	O
nonspecific	O
to	O
permit	O
unequivocal	O
confirmation	O
.	O

Thyroid	B-Gene_or_gene_product
transcription	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
TTF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
is	O
expressed	O
in	O
lung	B-Cancer
adenocarcinomas	I-Cancer
and	O
thyroid	B-Cancer
carcinomas	I-Cancer
but	O
not	O
in	O
adenocarcinomas	B-Cancer
arising	O
from	O
other	O
sites	B-Multi-tissue_structure
.	O

For	O
patients	B-Organism
with	O
adenocarcinomas	B-Cancer
in	O
the	O
lung	B-Organ
,	O
TTF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
staining	O
is	O
now	O
routinely	O
used	O
to	O
distinguish	O
a	O
primary	O
lung	B-Cancer
cancer	I-Cancer
from	O
a	O
lung	B-Organ
metastasis	O
.	O

Along	O
these	O
same	O
lines	O
,	O
TTF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
staining	O
might	O
prove	O
useful	O
in	O
localizing	O
the	O
tumor	B-Cancer
origin	O
of	O
adenocarcinomas	B-Cancer
encountered	O
outside	O
of	O
the	O
lung	B-Organ
.	O

The	O
archival	O
surgical	O
pathology	O
files	O
of	O
The	O
Johns	O
Hopkins	O
Hospital	O
were	O
searched	O
for	O
cases	O
of	O
brain	B-Organ
metastases	O
biopsied	O
between	O
1990	O
and	O
2000	O
.	O

Tissue	B-Tissue
blocks	I-Tissue
were	O
obtained	O
and	O
immunoperoxidase	O
staining	O
was	O
performed	O
using	O
the	O
TTF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
antibody	O
.	O

The	O
medical	O
records	O
were	O
reviewed	O
independent	O
of	O
the	O
staining	O
results	O
to	O
determine	O
site	O
of	O
tumor	B-Cancer
origin	O
.	O

Seventy	O
-	O
five	O
patients	B-Organism
underwent	O
biopsies	O
of	O
carcinomas	B-Cancer
metastatic	O
to	O
the	O
brain	B-Organ
.	O

At	O
the	O
time	O
of	O
brain	B-Organ
biopsy	O
,	O
the	O
primary	O
site	B-Multi-tissue_structure
of	O
tumor	B-Cancer
origin	O
was	O
known	O
in	O
45	O
cases	O
and	O
unknown	O
in	O
30	O
cases	O
.	O

Ultimately	O
,	O
the	O
primary	O
site	B-Multi-tissue_structure
was	O
established	O
on	O
clinical	O
and	O
radiographic	O
grounds	O
in	O
71	O
cases	O
(	O
95	O
%	O
)	O
.	O

These	O
included	O
40	O
(	O
56	O
%	O
)	O
metastases	O
from	O
a	O
primary	O
lung	B-Cancer
carcinoma	I-Cancer
and	O
31	O
(	O
44	O
%	O
)	O
metastases	O
from	O
some	O
nonpulmonary	B-Cancer
carcinoma	I-Cancer
.	O

TTF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
staining	O
was	O
present	O
in	O
31	O
of	O
the	O
40	O
(	O
78	O
%	O
)	O
metastatic	O
lung	B-Cancer
carcinomas	I-Cancer
,	O
but	O
in	O
only	O
1	O
of	O
the	O
31	O
(	O
3	O
%	O
)	O
metastatic	O
nonpulmonary	B-Cancer
carcinomas	I-Cancer
(	O
a	O
small	B-Cancer
-	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
of	O
the	O
sinonasal	B-Organism_subdivision
tract	I-Organism_subdivision
)	O
.	O

When	O
the	O
metastatic	O
lung	B-Cancer
carcinomas	I-Cancer
were	O
subtyped	O
,	O
TTF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
staining	O
was	O
noted	O
in	O
11	O
of	O
11	O
(	O
100	O
%	O
)	O
adenocarcinomas	B-Cancer
,	O
in	O
6	O
of	O
7	O
(	O
86	O
%	O
)	O
small	B-Cancer
-	I-Cancer
cell	I-Cancer
carcinomas	I-Cancer
,	O
in	O
15	O
of	O
19	O
(	O
79	O
%	O
)	O
large	B-Cancer
-	I-Cancer
cell	I-Cancer
carcinomas	I-Cancer
,	O
and	O
in	O
none	O
of	O
3	O
(	O
0	O
%	O
)	O
squamous	B-Cancer
cell	I-Cancer
carcinomas	I-Cancer
.	O

TTF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
staining	O
is	O
very	O
reliable	O
in	O
discerning	O
whether	O
a	O
brain	B-Organ
metastasis	O
has	O
arisen	O
from	O
a	O
pulmonary	B-Multi-tissue_structure
or	O
nonpulmonary	B-Multi-tissue_structure
site	I-Multi-tissue_structure
,	O
particularly	O
when	O
dealing	O
with	O
adenocarcinomas	B-Cancer
and	O
large	B-Cancer
-	I-Cancer
cell	I-Cancer
carcinomas	I-Cancer
.	O

TTF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
immunohistochemistry	O
could	O
focus	O
the	O
search	O
for	O
the	O
primary	O
tumor	B-Cancer
for	O
patients	B-Organism
presenting	O
with	O
brain	B-Organ
metastasis	O
as	O
the	O
initial	O
manifestation	O
.	O

Protein	O
expression	O
profile	O
of	O
primary	O
human	B-Organism
squamous	B-Cancer
cell	I-Cancer
lung	I-Cancer
carcinomas	I-Cancer
indicative	O
of	O
the	O
incidence	O
of	O
metastases	O
.	O

The	O
purpose	O
of	O
this	O
investigation	O
was	O
to	O
evaluate	O
firstly	O
whether	O
different	O
protein	O
expression	O
patterns	O
exist	O
in	O
primary	O
squamous	B-Cancer
cell	I-Cancer
lung	I-Cancer
carcinomas	I-Cancer
of	O
patients	B-Organism
with	O
and	O
without	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
involvement	O
and	O
secondly	O
,	O
whether	O
or	O
not	O
different	O
patterns	O
exist	O
in	O
tumours	B-Cancer
with	O
positive	O
lymph	B-Multi-tissue_structure
nodes	I-Multi-tissue_structure
.	O

For	O
this	O
reason	O
,	O
formalin	B-Simple_chemical
-	O
fixed	O
,	O
paraffin	B-Simple_chemical
-	O
embedded	O
specimens	B-Tissue
from	O
130	O
patients	B-Organism
with	O
squamous	B-Cancer
cell	I-Cancer
lung	I-Cancer
carcinomas	I-Cancer
were	O
analyzed	O
by	O
immunohistochemistry	O
.	O

In	O
a	O
first	O
step	O
,	O
proteins	O
were	O
selected	O
which	O
showed	O
a	O
relationship	O
to	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
involvement	O
.	O

The	O
expression	O
of	O
JUN	B-Gene_or_gene_product
,	O
ERBB2	B-Gene_or_gene_product
,	O
MYC	B-Gene_or_gene_product
,	O
cyclin	B-Gene_or_gene_product
D	I-Gene_or_gene_product
,	O
PCNA	B-Gene_or_gene_product
,	O
bFGF	B-Gene_or_gene_product
,	O
VEGF	B-Gene_or_gene_product
and	O
Hsp70	B-Gene_or_gene_product
proteins	O
revealed	O
a	O
positive	O
correlation	O
to	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
involvement	O
.	O

In	O
contrast	O
,	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
,	O
Fas	B-Gene_or_gene_product
ligand	I-Gene_or_gene_product
,	O
Fas	B-Gene_or_gene_product
/	O
CD95	B-Gene_or_gene_product
,	O
and	O
PAI	B-Gene_or_gene_product
showed	O
an	O
inverse	O
correlation	O
to	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
involvement	O
.	O

In	O
a	O
second	O
step	O
,	O
these	O
parameters	O
were	O
further	O
analyzed	O
by	O
hierarchical	O
cluster	O
analyses	O
.	O

The	O
resulting	O
clusters	O
were	O
correlated	O
to	O
patients	B-Organism
with	O
or	O
without	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
involvement	O
.	O

The	O
data	O
show	O
that	O
different	O
protein	O
expression	O
patterns	O
exist	O
between	O
primary	O
squamous	B-Cancer
cell	I-Cancer
lung	I-Cancer
carcinomas	I-Cancer
with	O
and	O
without	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
involvement	O
and	O
within	O
carcinomas	B-Cancer
with	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
involvement	O
.	O

The	O
data	O
suggest	O
that	O
various	O
metastasis	O
profiles	O
exist	O
.	O

Antiangiogenic	O
and	O
antitumor	B-Cancer
effects	O
of	O
a	O
protein	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
Cbeta	I-Gene_or_gene_product
inhibitor	O
in	O
human	B-Organism
breast	B-Cancer
cancer	I-Cancer
and	O
ovarian	B-Cancer
cancer	I-Cancer
xenografts	I-Cancer
.	O

In	O
cell	B-Cell
culture	O
,	O
the	O
compound	O
317615	B-Simple_chemical
2HCl	I-Simple_chemical
,	O
a	O
potent	O
inhibitor	O
of	O
VEGF	B-Gene_or_gene_product
-	O
stimulated	O
HUVEC	B-Cell
proliferation	O
,	O
was	O
not	O
very	O
effective	O
against	O
MX	B-Cell
-	I-Cell
1	I-Cell
breast	I-Cell
cancer	I-Cell
cells	I-Cell
(	O
IC50	O
=	O
8	O
.	O
1	O
microM	O
)	O
or	O
SKOV	B-Cell
-	I-Cell
3	I-Cell
ovarian	I-Cell
carcinoma	I-Cell
cells	I-Cell
(	O
IC50	O
=	O
9	O
.	O
5	O
microM	O
)	O
.	O

Exposure	O
to	O
combinations	O
of	O
paclitaxel	B-Simple_chemical
or	O
carboplatin	B-Simple_chemical
and	O
317615	B-Simple_chemical
x	I-Simple_chemical
2HCl	I-Simple_chemical
with	O
MX	B-Cell
-	I-Cell
1	I-Cell
cells	I-Cell
in	O
culture	O
resulted	O
in	O
cell	B-Cell
survival	O
that	O
reflected	O
primarily	O
additivity	O
of	O
the	O
two	O
agents	O
.	O

Exposure	O
of	O
SKOV	B-Cell
-	I-Cell
3	I-Cell
cells	I-Cell
to	O
paclitaxel	B-Simple_chemical
or	O
carboplatin	B-Simple_chemical
along	O
with	O
317615	B-Simple_chemical
2HCl	I-Simple_chemical
resulted	O
in	O
cell	B-Cell
survivals	O
that	O
reflected	O
additivity	O
of	O
317615	B-Simple_chemical
x	I-Simple_chemical
2HCl	I-Simple_chemical
with	O
paclitaxel	B-Simple_chemical
and	O
greater	O
-	O
than	O
-	O
additive	O
cytotoxicity	O
with	O
carboplatin	B-Simple_chemical
.	O

Administration	O
of	O
317615	B-Simple_chemical
x	I-Simple_chemical
2HCI	I-Simple_chemical
orally	O
twice	O
daily	O
to	O
nude	B-Organism
mice	I-Organism
bearing	O
subcutaneous	O
MX	B-Cancer
-	I-Cancer
1	I-Cancer
tumors	I-Cancer
or	O
SKOV	B-Cancer
-	I-Cancer
3	I-Cancer
tumors	I-Cancer
resulted	O
in	O
a	O
decreased	O
number	O
of	O
intratumoral	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
as	O
determined	O
by	O
CD31	B-Gene_or_gene_product
and	O
CD105	B-Gene_or_gene_product
staining	O
with	O
decreases	O
of	O
35	O
%	O
and	O
43	O
%	O
in	O
MX	B-Cancer
-	I-Cancer
1	I-Cancer
tumors	I-Cancer
and	O
60	O
%	O
and	O
75	O
%	O
in	O
SKOV	B-Cancer
-	I-Cancer
3	I-Cancer
tumors	I-Cancer
,	O
respectively	O
.	O

317615	B-Simple_chemical
x	I-Simple_chemical
2HCl	I-Simple_chemical
was	O
an	O
active	O
antitumor	B-Cancer
agent	O
against	O
the	O
MX	B-Cancer
-	I-Cancer
1	I-Cancer
xenograft	I-Cancer
and	O
increased	O
the	O
tumor	B-Cancer
growth	O
delay	O
produced	O
by	O
paclitaxel	B-Simple_chemical
by	O
1	O
.	O
7	O
-	O
fold	O
and	O
the	O
tumor	B-Cancer
growth	O
delay	O
produced	O
by	O
carboplatin	B-Simple_chemical
by	O
3	O
.	O
8	O
-	O
fold	O
.	O

Administration	O
of	O
317615	B-Simple_chemical
x	I-Simple_chemical
2HCl	I-Simple_chemical
also	O
increased	O
the	O
tumor	B-Cancer
growth	O
delay	O
produced	O
by	O
fractionated	O
radiation	O
therapy	O
in	O
the	O
MX	B-Cancer
-	I-Cancer
1	I-Cancer
tumor	I-Cancer
.	O

Treatment	O
with	O
317615	B-Simple_chemical
x	I-Simple_chemical
2HCl	I-Simple_chemical
alone	O
increased	O
the	O
lifespan	O
of	O
animals	O
bearing	O
intraperitoneal	B-Cancer
SKOV	I-Cancer
-	I-Cancer
3	I-Cancer
xenografts	I-Cancer
by	O
1	O
.	O
9	O
fold	O
compared	O
with	O
untreated	O
control	O
animals	O
.	O

The	O
combination	O
of	O
paclitaxel	B-Simple_chemical
and	O
317615	B-Simple_chemical
x	I-Simple_chemical
2HCl	I-Simple_chemical
resulted	O
in	O
100	O
%	O
120	O
-	O
day	O
survival	O
of	O
SKOV	B-Cancer
-	I-Cancer
3	I-Cancer
bearing	O
animals	O
.	O

Administration	O
of	O
317615	B-Simple_chemical
x	I-Simple_chemical
2HCl	I-Simple_chemical
along	O
with	O
carboplatin	B-Simple_chemical
to	O
animals	O
bearing	O
the	O
SKOV	B-Cancer
-	I-Cancer
3	I-Cancer
tumor	I-Cancer
produced	O
a	O
1	O
.	O
8	O
-	O
fold	O
increase	O
in	O
lifespan	O
compared	O
with	O
carboplatin	B-Simple_chemical
alone	O
.	O

317615	B-Simple_chemical
x	I-Simple_chemical
2HCl	I-Simple_chemical
is	O
a	O
promising	O
new	O
antiangiogenic	O
agent	O
that	O
is	O
in	O
early	O
phase	O
clinical	O
testing	O
.	O

Expression	O
of	O
CD154	B-Gene_or_gene_product
on	O
renal	B-Cancer
cell	I-Cancer
carcinomas	I-Cancer
and	O
effect	O
on	O
cell	B-Cell
proliferation	O
,	O
motility	O
and	O
platelet	B-Simple_chemical
-	I-Simple_chemical
activating	I-Simple_chemical
factor	I-Simple_chemical
synthesis	O
.	O

CD40	B-Gene_or_gene_product
activation	O
by	O
CD154	B-Gene_or_gene_product
may	O
trigger	O
diverse	O
cellular	B-Cell
responses	O
,	O
ranging	O
from	O
proliferation	O
and	O
differentiation	O
to	O
growth	O
suppression	O
and	O
cell	B-Cell
death	O
,	O
in	O
normal	O
and	O
malignant	B-Cell
cells	I-Cell
.	O

However	O
,	O
the	O
pathophysiologic	O
role	O
of	O
CD154	B-Gene_or_gene_product
expressed	O
by	O
tumor	B-Cell
cells	I-Cell
remains	O
unclear	O
.	O

We	O
have	O
investigated	O
the	O
expression	O
of	O
the	O
CD40	B-Gene_or_gene_product
-	O
CD154	B-Gene_or_gene_product
system	O
in	O
24	O
primary	O
cultures	O
derived	O
from	O
renal	B-Cancer
cell	I-Cancer
carcinomas	I-Cancer
,	O
its	O
correlation	O
with	O
tumor	B-Cancer
stage	O
and	O
its	O
potential	O
functional	O
significance	O
.	O

We	O
found	O
coexpression	O
of	O
CD40	B-Gene_or_gene_product
and	O
CD154	B-Gene_or_gene_product
in	O
most	O
of	O
the	O
renal	B-Cell
carcinoma	I-Cell
cell	I-Cell
lines	I-Cell
.	O

CD154	B-Gene_or_gene_product
,	O
but	O
not	O
CD40	B-Gene_or_gene_product
expression	O
,	O
significantly	O
correlated	O
with	O
tumor	B-Cancer
stage	O
.	O

Moreover	O
,	O
renal	B-Cell
carcinoma	I-Cell
cell	I-Cell
lines	I-Cell
also	O
released	O
the	O
soluble	O
form	O
of	O
CD154	B-Gene_or_gene_product
into	O
the	O
supernatant	O
.	O

CD40	B-Gene_or_gene_product
engagement	O
by	O
CD154	B-Gene_or_gene_product
did	O
not	O
affect	O
apoptosis	O
or	O
survival	O
.	O

On	O
the	O
contrary	O
,	O
CD154	B-Gene_or_gene_product
stimulated	O
cell	B-Cell
proliferation	O
,	O
motility	O
and	O
production	O
of	O
PAF	B-Simple_chemical
,	O
a	O
phospholipid	O
mediator	O
of	O
inflammation	O
with	O
angiogenic	O
properties	O
.	O

Furthermore	O
,	O
the	O
renal	B-Cell
carcinoma	I-Cell
cell	I-Cell
lines	I-Cell
expressed	O
PAF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
R	I-Gene_or_gene_product
.	O

Blockade	O
of	O
PAF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
R	I-Gene_or_gene_product
by	O
WEB	B-Simple_chemical
-	I-Simple_chemical
2170	I-Simple_chemical
,	O
a	O
PAF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
R	I-Gene_or_gene_product
antagonist	O
,	O
abolished	O
the	O
CD154	B-Gene_or_gene_product
-	O
dependent	O
motility	O
,	O
indicating	O
a	O
role	O
for	O
PAF	B-Simple_chemical
synthesized	O
after	O
CD154	B-Gene_or_gene_product
stimulation	O
in	O
renal	B-Cell
carcinoma	I-Cell
cell	I-Cell
motility	O
.	O

In	O
conclusion	O
,	O
this	O
study	O
identifies	O
new	O
functional	O
properties	O
for	O
CD154	B-Gene_or_gene_product
,	O
which	O
are	O
potentially	O
relevant	O
for	O
the	O
growth	O
and	O
dissemination	O
of	O
renal	B-Cell
carcinoma	I-Cell
cells	I-Cell
.	O

DNA	B-Cellular_component
damage	O
induces	O
a	O
novel	O
p53	B-Gene_or_gene_product
-	O
survivin	B-Gene_or_gene_product
signaling	O
pathway	O
regulating	O
cell	B-Cell
cycle	O
and	O
apoptosis	O
in	O
acute	B-Cell
lymphoblastic	I-Cell
leukemia	I-Cell
cells	I-Cell
.	O

Survivin	B-Gene_or_gene_product
is	O
a	O
novel	O
member	O
of	O
the	O
inhibitor	B-Gene_or_gene_product
of	I-Gene_or_gene_product
apoptosis	I-Gene_or_gene_product
protein	O
(	O
IAP	B-Gene_or_gene_product
)	O
family	O
.	O

Here	O
we	O
report	O
that	O
the	O
chemotherapeutic	O
drug	O
doxorubicin	B-Simple_chemical
,	O
a	O
DNA	B-Cellular_component
-	O
damaging	O
agent	O
,	O
activates	O
a	O
p53	B-Gene_or_gene_product
-	O
survivin	B-Gene_or_gene_product
signaling	O
pathway	O
inducing	O
cell	B-Cell
cycle	O
arrest	O
and	O
apoptosis	O
in	O
childhood	O
acute	B-Cancer
lymphoblastic	I-Cancer
leukemia	I-Cancer
(	O
ALL	B-Cancer
)	O
.	O

Treatment	O
of	O
wild	O
-	O
type	O
(	O
wt	O
)	O
p53	B-Gene_or_gene_product
ALL	O
cells	O
(	O
EU	B-Cell
-	I-Cell
3	I-Cell
cell	I-Cell
line	I-Cell
)	O
with	O
doxorubicin	B-Simple_chemical
caused	O
accumulation	O
of	O
p53	B-Gene_or_gene_product
,	O
resulting	O
in	O
dramatic	O
down	O
-	O
regulation	O
of	O
survivin	B-Gene_or_gene_product
,	O
depletion	O
of	O
cells	B-Cell
in	O
G	O
(	O
2	O
)	O
/	O
M	O
,	O
and	O
apoptosis	O
(	O
increased	O
sub	O
-	O
G	O
(	O
1	O
)	O
compartment	O
)	O
.	O

In	O
contrast	O
,	O
doxorubicin	B-Simple_chemical
treatment	O
of	O
mutant	O
(	O
mut	O
)	O
p53	B-Gene_or_gene_product
cells	O
(	O
EU	B-Cell
-	I-Cell
6	I-Cell
/	I-Cell
ALL	I-Cell
line	I-Cell
)	O
up	O
-	O
regulated	O
survivin	B-Gene_or_gene_product
and	O
induced	O
G	O
(	O
2	O
)	O
/	O
M	O
arrest	O
without	O
inducing	O
apoptosis	O
.	O

However	O
,	O
treating	O
EU	B-Cell
-	I-Cell
6	I-Cell
with	O
anti	O
-	O
survivin	B-Gene_or_gene_product
antisense	O
resensitized	O
these	O
cells	B-Cell
to	O
doxorubicin	B-Simple_chemical
,	O
resulting	O
in	O
apoptosis	O
.	O

With	O
a	O
p53	B-Gene_or_gene_product
-	O
null	O
cell	O
line	O
(	O
EU	B-Cell
-	I-Cell
4	I-Cell
)	I-Cell
,	O
although	O
doxorubicin	B-Simple_chemical
treatment	O
arrested	O
cells	B-Cell
in	O
G	O
(	O
2	O
)	O
/	O
M	O
,	O
survivin	B-Gene_or_gene_product
expression	O
was	O
unchanged	O
,	O
and	O
cells	B-Cell
underwent	O
only	O
limited	O
apoptosis	O
.	O

However	O
,	O
re	O
-	O
expression	O
of	O
wt	O
-	O
p53	B-Gene_or_gene_product
in	O
EU	B-Cell
-	I-Cell
4	I-Cell
cells	I-Cell
could	O
restore	O
the	O
doxorubicin	B-Simple_chemical
-	O
p53	B-Gene_or_gene_product
-	O
survivin	B-Gene_or_gene_product
pathway	O
,	O
resulting	O
in	O
significantly	O
decreased	O
survivin	B-Gene_or_gene_product
expression	O
and	O
increased	O
apoptosis	O
in	O
these	O
cells	B-Cell
after	O
doxorubicin	B-Simple_chemical
treatment	O
.	O

Following	O
cotransfection	O
of	O
p53	B-Gene_or_gene_product
-	O
null	O
EU	O
-	O
4	O
cells	O
with	O
survivin	B-Gene_or_gene_product
promoter	O
-	O
luciferase	B-Gene_or_gene_product
constructs	O
and	O
either	O
wt	O
-	O
p53	B-Gene_or_gene_product
or	O
different	O
mut	O
-	O
p53	B-Gene_or_gene_product
expression	O
vectors	O
,	O
wt	O
-	O
p53	B-Gene_or_gene_product
inhibited	O
survivin	B-Gene_or_gene_product
promoter	O
activity	O
;	O
p53	B-Gene_or_gene_product
-	O
mediated	O
inhibition	O
could	O
be	O
abrogated	O
by	O
overexpression	O
of	O
murine	B-Gene_or_gene_product
double	I-Gene_or_gene_product
minute2	I-Gene_or_gene_product
(	O
MDM2	B-Gene_or_gene_product
)	O
protein	O
.	O

Together	O
,	O
these	O
studies	O
define	O
a	O
novel	O
p53	B-Gene_or_gene_product
-	O
survivin	B-Gene_or_gene_product
signaling	O
pathway	O
activated	O
by	O
DNA	B-Cellular_component
damage	O
that	O
results	O
in	O
down	O
-	O
regulation	O
of	O
survivin	B-Gene_or_gene_product
,	O
cell	B-Cell
cycle	O
arrest	O
,	O
and	O
apoptosis	O
.	O

Furthermore	O
,	O
our	O
data	O
indicate	O
that	O
loss	O
of	O
wt	O
-	O
p53	B-Gene_or_gene_product
function	O
in	O
tumor	B-Cell
cells	I-Cell
may	O
contribute	O
to	O
up	O
-	O
regulation	O
of	O
survivin	B-Gene_or_gene_product
and	O
resistance	O
to	O
DNA	B-Cellular_component
-	O
damaging	O
agents	O
.	O

Effects	O
of	O
4	B-Simple_chemical
-	I-Simple_chemical
alkylmorpholine	I-Simple_chemical
N	I-Simple_chemical
-	I-Simple_chemical
oxides	I-Simple_chemical
on	O
ATP	B-Simple_chemical
-	O
producing	O
processes	O
in	O
Ehrlich	B-Cell
ascites	I-Cell
and	O
L1210	B-Cell
leukaemia	I-Cell
cells	I-Cell
.	O

The	O
main	O
purpose	O
of	O
the	O
present	O
investigation	O
was	O
to	O
study	O
the	O
effect	O
of	O
the	O
homologous	O
series	O
of	O
4	B-Simple_chemical
-	I-Simple_chemical
alkylmorpholine	I-Simple_chemical
N	I-Simple_chemical
-	I-Simple_chemical
oxides	I-Simple_chemical
on	O
ATP	B-Simple_chemical
-	O
producing	O
processes	O
in	O
Ehrlich	B-Cell
ascites	I-Cell
and	O
L1210	B-Cell
murine	I-Cell
leukaemia	I-Cell
cells	I-Cell
.	O

The	O
effects	O
on	O
aerobic	O
glucose	B-Simple_chemical
consumption	O
,	O
lactic	B-Simple_chemical
acid	I-Simple_chemical
formation	O
,	O
content	O
of	O
total	O
(	O
T	O
-	O
SH	O
)	O
and	O
non	O
-	O
protein	O
thiol	O
groups	O
(	O
NP	O
-	O
SH	O
)	O
,	O
endogenous	O
and	O
exogenous	O
respiration	O
and	O
the	O
level	O
of	O
ATP	B-Simple_chemical
in	O
tumour	B-Cell
cells	I-Cell
incubated	O
in	O
vitro	O
were	O
investigated	O
.	O

4	B-Simple_chemical
-	I-Simple_chemical
Dodecylmorpholine	I-Simple_chemical
N	I-Simple_chemical
-	I-Simple_chemical
oxide	I-Simple_chemical
(	O
DMNO	B-Simple_chemical
)	O
,	O
one	O
of	O
the	O
most	O
active	O
compounds	O
,	O
decreased	O
the	O
level	O
of	O
ATP	B-Simple_chemical
immediately	O
after	O
addition	O
to	O
the	O
suspension	O
of	O
Ehrlich	B-Cell
cells	I-Cell
in	O
an	O
ice	O
bath	O
.	O

After	O
2	O
h	O
incubation	O
at	O
37	O
degrees	O
C	O
the	O
drop	O
in	O
the	O
ATP	B-Simple_chemical
level	O
was	O
much	O
lower	O
.	O

A	O
possible	O
explanation	O
for	O
the	O
decrease	O
in	O
the	O
ATP	B-Simple_chemical
level	O
might	O
be	O
interaction	O
of	O
the	O
amine	B-Simple_chemical
oxide	I-Simple_chemical
with	O
the	O
cell	B-Cellular_component
membrane	I-Cellular_component
.	O

Fibroblast	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
2	I-Gene_or_gene_product
promotes	O
microvessel	B-Tissue
formation	O
from	O
mouse	B-Organism
embryonic	B-Multi-tissue_structure
aorta	I-Multi-tissue_structure
.	O

To	O
delineate	O
the	O
roles	O
that	O
oxygen	B-Simple_chemical
and	O
fibroblast	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factors	I-Gene_or_gene_product
(	O
FGFs	B-Gene_or_gene_product
)	O
play	O
in	O
the	O
process	O
of	O
angiogenesis	O
from	O
the	O
embryonic	B-Multi-tissue_structure
aorta	I-Multi-tissue_structure
,	O
we	O
cultured	O
mouse	B-Organism
embryonic	B-Multi-tissue_structure
aorta	I-Multi-tissue_structure
explants	I-Multi-tissue_structure
(	O
thoracic	B-Multi-tissue_structure
level	O
to	O
lateral	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
supplying	O
the	O
mesonephros	B-Organ
and	O
metanephros	B-Organ
)	O
in	O
a	O
three	O
-	O
dimensional	O
type	B-Gene_or_gene_product
I	I-Gene_or_gene_product
collagen	I-Gene_or_gene_product
gel	O
matrix	O
.	O

During	O
8	O
days	O
of	O
culture	O
under	O
5	O
%	O
O	B-Simple_chemical
(	I-Simple_chemical
2	I-Simple_chemical
)	I-Simple_chemical
,	O
but	O
not	O
room	O
air	O
,	O
the	O
addition	O
of	O
FGF2	B-Gene_or_gene_product
to	O
explants	O
stimulated	O
the	O
formation	O
of	O
Gs	B-Gene_or_gene_product
-	I-Gene_or_gene_product
IB	I-Gene_or_gene_product
(	I-Gene_or_gene_product
4	I-Gene_or_gene_product
-	I-Gene_or_gene_product
)	I-Gene_or_gene_product
positive	O
,	O
CD31	B-Gene_or_gene_product
-	O
positive	O
,	O
and	O
Flk	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
positive	O
microvessels	O
in	O
a	O
concentration	O
-	O
dependent	O
manner	O
.	O

FGF2	B-Gene_or_gene_product
-	O
stimulated	O
microvessel	B-Tissue
formation	O
was	O
inhibited	O
by	O
sequestration	O
of	O
FGF2	B-Gene_or_gene_product
via	O
addition	O
of	O
soluble	O
FGF	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
(	O
FGFR	B-Gene_or_gene_product
)	O
chimera	O
protein	O
or	O
anti	O
-	O
FGF2	B-Gene_or_gene_product
antibodies	O
.	O

FGFR1	B-Gene_or_gene_product
and	O
FGFR2	B-Gene_or_gene_product
were	O
present	O
on	O
explants	O
.	O

Levels	O
of	O
FGFR1	B-Gene_or_gene_product
,	O
but	O
not	O
FGFR2	B-Gene_or_gene_product
,	O
were	O
increased	O
in	O
embryonic	B-Multi-tissue_structure
aorta	I-Multi-tissue_structure
cultured	O
under	O
5	O
%	O
O	B-Simple_chemical
(	I-Simple_chemical
2	I-Simple_chemical
)	I-Simple_chemical
relative	O
to	O
room	O
air	O
.	O

Our	O
data	O
suggest	O
that	O
low	O
oxygen	B-Simple_chemical
upregulates	O
FGFR1	B-Gene_or_gene_product
expression	O
in	O
embryonic	B-Multi-tissue_structure
aorta	I-Multi-tissue_structure
in	O
vitro	O
and	O
renders	O
it	O
more	O
responsive	O
to	O
FGF2	B-Gene_or_gene_product
.	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
I	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
-	O
mediated	O
vasculogenesis	O
/	O
angiogenesis	O
in	O
human	B-Organism
lung	B-Organ
development	O
.	O

The	O
structural	O
and	O
functional	O
development	O
of	O
the	O
pulmonary	B-Anatomical_system
system	I-Anatomical_system
is	O
dependent	O
upon	O
appropriate	O
early	O
vascularization	O
of	O
the	O
embryonic	B-Organ
lung	I-Organ
.	O

Our	O
previous	O
in	O
vitro	O
studies	O
in	O
a	O
rat	B-Organism
model	O
indicated	O
that	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
I	I-Gene_or_gene_product
(	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
I	I-Gene_or_gene_product
)	O
is	O
a	O
potent	O
angiogenic	O
agent	O
for	O
fetal	B-Cell
lung	I-Cell
endothelial	I-Cell
cells	I-Cell
.	O

To	O
assess	O
its	O
role	O
on	O
human	B-Organism
vascular	B-Multi-tissue_structure
lung	B-Organ
development	O
,	O
we	O
first	O
examined	O
the	O
expression	O
of	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
I	I-Gene_or_gene_product
/	O
II	B-Gene_or_gene_product
and	O
IGF	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
type	I-Gene_or_gene_product
I	I-Gene_or_gene_product
(	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
IR	I-Gene_or_gene_product
)	O
in	O
human	B-Organism
embryonic	B-Tissue
and	O
fetal	B-Tissue
lung	I-Tissue
tissues	I-Tissue
at	O
4	O
-	O
12	O
wk	O
of	O
gestation	O
.	O

Immunohistochemical	O
and	O
in	O
situ	O
hybridization	O
studies	O
revealed	O
the	O
presence	O
of	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
I	I-Gene_or_gene_product
/	O
II	B-Gene_or_gene_product
-	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
IR	I-Gene_or_gene_product
ligands	O
and	O
mRNA	O
transcripts	O
in	O
embryonic	B-Organ
lungs	I-Organ
as	O
early	O
as	O
4	O
wk	O
gestation	O
.	O

Immunotargeting	O
using	O
an	O
anti	O
-	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
IR	I-Gene_or_gene_product
neutralizing	O
antibody	O
on	O
human	B-Organism
fetal	B-Organ
lung	I-Organ
explants	I-Organ
demonstrated	O
a	O
significant	O
blockade	O
of	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
IR	I-Gene_or_gene_product
signaling	O
.	O

Inactivation	O
of	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
IR	I-Gene_or_gene_product
resulted	O
in	O
a	O
loss	O
of	O
endothelial	B-Cell
cells	I-Cell
,	O
accompanied	O
by	O
dramatic	O
changes	O
in	O
fetal	B-Organ
lung	I-Organ
explant	O
morphology	O
.	O

Terminal	O
transferase	O
dUTP	O
end	O
-	O
labeling	O
assay	O
and	O
TEM	O
studies	O
of	O
anti	O
-	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
IR	I-Gene_or_gene_product
-	O
treated	O
lungs	B-Organ
demonstrated	O
numerous	O
apoptotic	O
mesenchymal	B-Cell
cells	I-Cell
.	O

Rat	B-Organism
embryonic	B-Organ
lung	I-Organ
explant	O
studies	O
further	O
validated	O
the	O
importance	O
of	O
the	O
IGF	B-Gene_or_gene_product
-	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
IR	I-Gene_or_gene_product
system	O
for	O
lung	B-Multi-tissue_structure
vascular	I-Multi-tissue_structure
development	O
.	O

These	O
data	O
provide	O
the	O
first	O
demonstration	O
of	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
I	I-Gene_or_gene_product
/	O
II	B-Gene_or_gene_product
expression	O
in	O
the	O
human	B-Organism
lung	B-Organ
in	O
early	O
gestation	O
and	O
indicate	O
that	O
the	O
IGF	B-Gene_or_gene_product
family	O
of	O
growth	O
factors	O
,	O
acting	O
through	O
the	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
IR	I-Gene_or_gene_product
,	O
is	O
required	O
as	O
a	O
survival	O
factor	O
during	O
normal	O
human	B-Organism
lung	B-Organ
vascularization	O
.	O

Enhanced	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
by	O
periodontal	B-Organism_subdivision
pathogens	O
in	O
gingival	B-Cell
fibroblasts	I-Cell
.	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
has	O
recently	O
attracted	O
attention	O
as	O
a	O
potent	O
inducer	O
of	O
vascular	B-Multi-tissue_structure
permeability	O
and	O
angiogenesis	O
.	O

Aberrant	O
angiogenesis	O
is	O
often	O
associated	O
with	O
lesion	B-Pathological_formation
formation	O
in	O
chronic	O
periodontitis	O
.	O

The	O
aim	O
of	O
the	O
present	O
study	O
was	O
to	O
investigate	O
the	O
properties	O
of	O
VEGF	B-Gene_or_gene_product
expression	O
in	O
human	B-Organism
gingival	B-Cell
fibroblasts	I-Cell
(	O
HGF	B-Cell
)	O
culture	O
.	O

HGF	B-Cell
were	O
stimulated	O
with	O
lipopolysaccharide	B-Simple_chemical
(	O
LPS	B-Simple_chemical
)	O
,	O
vesicle	B-Gene_or_gene_product
(	O
Ve	B-Gene_or_gene_product
)	O
and	O
outer	B-Gene_or_gene_product
membrane	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
(	O
OMP	B-Gene_or_gene_product
)	O
from	O
Actinobacillus	B-Organism
actinomycetemcomitans	I-Organism
and	O
Porphyromonas	B-Organism
gingivalis	I-Organism
.	O

HGF	B-Cell
constitutively	O
produced	O
VEGF	B-Gene_or_gene_product
and	O
levels	O
were	O
significantly	O
enhanced	O
(	O
P	O
less	O
than	O
0	O
.	O
01	O
)	O
by	O
stimulation	O
with	O
Ve	B-Gene_or_gene_product
and	O
OMP	B-Gene_or_gene_product
from	O
A	B-Organism
.	I-Organism
actinomycetemcomitans	I-Organism
and	O
P	B-Organism
.	I-Organism
gingivalis	I-Organism
at	O
concentrations	O
of	O
10	O
microg	O
/	O
ml	O
or	O
higher	O
.	O

On	O
the	O
other	O
hand	O
,	O
VEGF	B-Gene_or_gene_product
levels	O
were	O
not	O
increased	O
by	O
LPS	B-Simple_chemical
stimulation	O
.	O

VEGF	B-Gene_or_gene_product
mRNA	O
expression	O
was	O
also	O
observed	O
in	O
Ve	B-Gene_or_gene_product
-	O
and	O
OMP	B-Gene_or_gene_product
-	O
stimulated	O
HGF	B-Cell
.	O

A	O
vascular	B-Multi-tissue_structure
permeability	O
enhancement	O
(	O
VPE	O
)	O
assay	O
was	O
performed	O
using	O
guinea	B-Organism
pigs	I-Organism
to	O
ascertain	O
whether	O
supernatant	B-Organism_substance
from	O
cultures	O
of	O
Ve	B-Gene_or_gene_product
-	O
and	O
OMP	B-Gene_or_gene_product
-	O
stimulated	O
HGF	B-Cell
enhance	O
vascular	B-Multi-tissue_structure
permeability	O
in	O
vivo	O
.	O

Supernatant	O
from	O
cultures	O
of	O
Ve	B-Gene_or_gene_product
-	O
and	O
OMP	B-Gene_or_gene_product
-	O
stimulated	O
HGF	B-Cell
strongly	O
induced	O
VPE	O
.	O

This	O
was	O
markedly	O
suppressed	O
upon	O
simultaneous	O
injection	O
of	O
anti	O
-	O
VEGF	B-Gene_or_gene_product
polyclonal	O
antibodies	O
with	O
the	O
supernatant	O
.	O

Heating	O
and	O
protease	O
treatment	O
of	O
the	O
stimulants	O
reduced	O
the	O
VEGF	B-Gene_or_gene_product
enhancing	O
levels	O
in	O
Ve	B-Gene_or_gene_product
and	O
OMP	B-Gene_or_gene_product
in	O
vitro	O
.	O

These	O
results	O
suggest	O
that	O
Ve	B-Gene_or_gene_product
and	O
OMP	B-Gene_or_gene_product
may	O
be	O
crucial	O
heat	O
-	O
labile	O
and	O
protease	O
-	O
sensitive	O
components	O
of	O
periodontal	B-Organism_subdivision
pathogens	O
that	O
enhance	O
VEGF	B-Gene_or_gene_product
expression	O
.	O

In	O
addition	O
,	O
VEGF	B-Gene_or_gene_product
might	O
be	O
associated	O
with	O
the	O
etiology	O
of	O
periodontitis	O
in	O
its	O
early	O
stages	O
according	O
to	O
neovascularization	O
stimulated	O
by	O
periodontal	B-Organism_subdivision
pathogens	O
causing	O
swelling	O
and	O
edema	B-Pathological_formation
.	O

NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
/	O
Rel	B-Gene_or_gene_product
transcriptional	O
pathway	O
:	O
implications	O
in	O
pancreatic	B-Cancer
cancer	I-Cancer
.	O

Despite	O
considerable	O
efforts	O
in	O
understanding	O
the	O
cellular	B-Cell
mechanisms	O
contributing	O
to	O
pancreatic	B-Cancer
cancer	I-Cancer
,	O
the	O
prognosis	O
of	O
this	O
malignant	O
disease	O
is	O
still	O
extremely	O
poor	O
.	O

Although	O
pancreatic	B-Cancer
cancer	I-Cancer
is	O
the	O
fifth	O
common	O
cause	O
of	O
cancer	B-Cancer
death	O
in	O
Western	O
countries	O
,	O
current	O
options	O
in	O
treatment	O
enable	O
a	O
5	O
-	O
yr	O
survival	O
rate	O
for	O
all	O
stages	O
of	O
less	O
than	O
5	O
%	O
.	O

In	O
the	O
face	O
fo	O
the	O
fatal	B-Developing_anatomical_structure
outcome	O
,	O
new	O
approaches	O
to	O
the	O
therapy	O
have	O
been	O
established	O
.	O

Based	O
on	O
its	O
role	O
in	O
malignant	O
transformation	O
,	O
apoptosis	O
,	O
and	O
cell	B-Cell
proliferation	O
,	O
the	O
transcription	O
factor	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
/	O
Rel	B-Gene_or_gene_product
has	O
gained	O
the	O
attention	O
of	O
many	O
laboratories	O
.	O

This	O
review	O
provides	O
basic	O
information	O
for	O
the	O
understanding	O
of	O
the	O
biology	O
of	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
and	O
aims	O
at	O
presenting	O
experimental	O
data	O
illustrating	O
the	O
involvement	O
of	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
/	O
Rel	B-Gene_or_gene_product
in	O
pancreatic	B-Cancer
cancer	I-Cancer
.	O

Combined	O
topical	O
fluconazole	B-Simple_chemical
and	O
corticosteroid	B-Simple_chemical
treatment	O
for	O
experimental	O
Candida	B-Organism
albicans	I-Organism
keratomycosis	O
.	O

PURPOSE	O
:	O
To	O
determine	O
the	O
most	O
efficient	O
time	O
point	O
and	O
concentration	O
of	O
topical	O
corticosteroids	B-Simple_chemical
in	O
Candida	B-Organism
albicans	I-Organism
keratitis	O
treated	O
with	O
fluconazole	B-Simple_chemical
.	O

METHODS	O
:	O
Corneas	B-Multi-tissue_structure
of	O
105	O
rabbits	B-Organism
were	O
infected	O
with	O
viable	O
yeast	B-Organism
cells	B-Cell
of	O
C	B-Organism
.	I-Organism
albicans	I-Organism
(	O
2	O
.	O
5	O
x	O
10	O
(	O
5	O
)	O
)	O
.	O

After	O
a	O
48	O
-	O
hour	O
incubation	O
period	O
,	O
seven	O
groups	O
of	O
animals	O
were	O
treated	O
for	O
21	O
days	O
with	O
fluconazole	B-Simple_chemical
,	O
with	O
group	O
I	O
acting	O
as	O
a	O
control	O
,	O
and	O
groups	O
II	O
to	O
VII	O
receiving	O
adjunct	O
therapy	O
with	O
the	O
corticosteroid	B-Simple_chemical
prednisolone	B-Simple_chemical
(	O
5	O
or	O
10	O
times	O
daily	O
;	O
3	O
,	O
9	O
,	O
or	O
15	O
days	O
after	O
infection	O
)	O
.	O

The	O
degree	O
of	O
corneal	B-Multi-tissue_structure
infiltration	O
,	O
ulceration	O
,	O
corneal	B-Multi-tissue_structure
clouding	O
,	O
hypopyon	O
,	O
conjunctivitis	O
,	O
neovascularization	O
,	O
and	O
corneal	B-Multi-tissue_structure
perforation	O
was	O
monitored	O
over	O
a	O
24	O
-	O
day	O
period	O
,	O
as	O
well	O
as	O
recultivation	O
and	O
resistance	O
to	O
fluconazole	B-Simple_chemical
of	O
the	O
C	B-Organism
.	I-Organism
albicans	I-Organism
pathogen	O
.	O

RESULTS	O
:	O
The	O
control	O
group	O
showed	O
the	O
highest	O
level	O
of	O
corneal	B-Multi-tissue_structure
clouding	O
and	O
neovascularization	O
.	O

In	O
comparison	O
,	O
by	O
day	O
24	O
,	O
the	O
majority	O
of	O
groups	O
also	O
treated	O
with	O
prednisolone	B-Simple_chemical
displayed	O
significantly	O
less	O
corneal	B-Multi-tissue_structure
clouding	O
and	O
neovascularization	O
.	O

An	O
immediate	O
decrease	O
in	O
corneal	B-Multi-tissue_structure
clouding	O
was	O
observed	O
in	O
groups	O
treated	O
with	O
additional	O
low	O
-	O
or	O
high	O
-	O
dose	O
prednisolone	B-Simple_chemical
from	O
day	O
9	O
after	O
inoculation	O
.	O

After	O
additional	O
prednisolone	B-Simple_chemical
treatment	O
from	O
day	O
9	O
or	O
15	O
after	O
inoculation	O
,	O
no	O
significant	O
difference	O
was	O
detected	O
in	O
the	O
recultivation	O
rate	O
of	O
C	B-Organism
.	I-Organism
albicans	I-Organism
compared	O
with	O
the	O
control	O
.	O

Early	O
administration	O
of	O
prednisolone	B-Simple_chemical
(	O
day	O
3	O
,	O
low	O
and	O
high	O
dose	O
)	O
resulted	O
in	O
the	O
recultivation	O
of	O
significantly	O
more	O
C	B-Organism
.	I-Organism
albicans	I-Organism
.	O

CONCLUSIONS	O
:	O
Fluconazole	B-Simple_chemical
plus	O
adjunct	O
high	O
-	O
dose	O
prednisolone	B-Simple_chemical
treatment	O
was	O
most	O
effective	O
when	O
administered	O
9	O
days	O
after	O
infection	O
.	O

The	O
delayed	O
application	O
of	O
corticosteroids	B-Simple_chemical
after	O
treatment	O
with	O
antimycotic	B-Simple_chemical
drugs	I-Simple_chemical
in	O
cases	O
of	O
fungal	O
keratitis	O
is	O
therefore	O
not	O
contraindicated	O
and	O
may	O
be	O
beneficial	O
in	O
patients	B-Organism
.	O

Solid	B-Cancer
tumor	I-Cancer
therapy	O
:	O
manipulation	O
of	O
the	O
vasculature	B-Multi-tissue_structure
with	O
TNF	B-Gene_or_gene_product
.	O

Drug	O
delivery	O
to	O
solid	B-Cancer
tumors	I-Cancer
is	O
one	O
of	O
the	O
most	O
challenging	O
aspects	O
in	O
cancer	B-Cancer
therapy	O
.	O

Whereas	O
agents	O
seem	O
promising	O
in	O
the	O
test	O
tube	O
,	O
clinical	O
trials	O
often	O
fail	O
due	O
to	O
unfavorable	O
pharmacokinetics	O
,	O
poor	O
delivery	O
,	O
low	O
local	O
concentrations	O
,	O
and	O
limited	O
accumulation	O
in	O
the	O
target	O
cell	B-Cell
.	O

A	O
major	O
step	O
forwards	O
in	O
the	O
treatment	O
of	O
solid	B-Cancer
tumors	I-Cancer
is	O
the	O
recognition	O
of	O
the	O
tumor	B-Cancer
-	O
associated	O
vasculature	B-Multi-tissue_structure
as	O
an	O
important	O
target	O
for	O
therapy	O
.	O

Inhibition	O
of	O
tumor	B-Multi-tissue_structure
vascular	I-Multi-tissue_structure
development	O
has	O
a	O
direct	O
effect	O
on	O
the	O
growth	O
and	O
progression	O
of	O
the	O
tumor	B-Cancer
.	O

Destruction	O
of	O
an	O
existing	O
vasculature	B-Multi-tissue_structure
also	O
directly	O
inflicts	O
serious	O
damage	O
to	O
the	O
tumor	B-Cell
cell	I-Cell
.	O

Moreover	O
,	O
the	O
tumor	B-Multi-tissue_structure
vascular	I-Multi-tissue_structure
bed	I-Multi-tissue_structure
can	O
be	O
manipulated	O
facilitating	O
enhanced	O
permissiveness	O
of	O
the	O
tumor	B-Cancer
for	O
administered	O
chemotherapeutics	O
.	O

In	O
this	O
review	O
,	O
we	O
focus	O
on	O
the	O
use	O
of	O
tumor	B-Gene_or_gene_product
necrosis	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
(	O
TNF	B-Gene_or_gene_product
)	O
in	O
local	O
and	O
systemic	O
therapy	O
in	O
conjunction	O
with	O
chemotherapy	O
.	O

In	O
these	O
settings	O
TNF	B-Gene_or_gene_product
demonstrates	O
potent	O
and	O
selective	O
activity	O
on	O
the	O
tumor	B-Multi-tissue_structure
vascular	I-Multi-tissue_structure
bed	I-Multi-tissue_structure
,	O
which	O
strongly	O
improves	O
tumor	B-Cancer
response	O
.	O

Comparison	O
of	O
tissue	B-Tissue
integration	O
between	O
polyester	B-Simple_chemical
and	O
polypropylene	B-Simple_chemical
prostheses	O
in	O
the	O
preperitoneal	B-Immaterial_anatomical_entity
space	I-Immaterial_anatomical_entity
.	O

Tissue	B-Tissue
integration	O
and	O
implant	O
characteristics	O
of	O
various	O
biomaterials	O
commonly	O
used	O
for	O
inguinal	B-Pathological_formation
hernia	I-Pathological_formation
repair	O
have	O
not	O
been	O
studied	O
extensively	O
.	O

The	O
aim	O
of	O
this	O
study	O
is	O
to	O
compare	O
behavior	O
and	O
tissue	B-Tissue
response	O
between	O
two	O
new	O
polyester	B-Simple_chemical
prostheses	O
and	O
a	O
commonly	O
used	O
polypropylene	B-Simple_chemical
(	O
PP	B-Simple_chemical
)	O
mesh	O
.	O

The	O
polyester	B-Simple_chemical
prostheses	O
utilized	O
were	O
polyester	B-Simple_chemical
flat	O
(	O
PF	O
)	O
and	O
polyester	B-Simple_chemical
soft	O
three	O
-	O
dimensional	O
(	O
PS	O
)	O
;	O
the	O
PP	O
mesh	O
utilized	O
was	O
Marlex	O
.	O

Eight	O
randomly	O
assigned	O
4	O
x	O
4	O
-	O
cm2	O
pieces	O
of	O
two	O
different	O
meshes	O
were	O
fixed	O
in	O
the	O
preperitoneal	B-Immaterial_anatomical_entity
space	I-Immaterial_anatomical_entity
with	O
a	O
centrally	O
placed	O
single	O
suture	O
.	O

Gross	O
evaluation	O
included	O
shrinkage	O
and	O
stiffness	O
.	O

Histological	O
evaluation	O
included	O
amount	O
of	O
fibrous	B-Tissue
and	O
fat	B-Tissue
encapsulation	O
,	O
connective	B-Tissue
tissue	I-Tissue
,	O
foreign	O
-	O
body	O
reaction	O
,	O
neovascularization	O
,	O
hemorrhage	O
,	O
necrosis	O
,	O
and	O
exudate	B-Organism_substance
.	O

Evaluations	O
were	O
graded	O
on	O
a	O
zero	O
to	O
four	O
scale	O
.	O

The	O
area	O
and	O
the	O
area	O
ratio	O
were	O
measured	O
using	O
a	O
calibrated	O
micrometer	O
.	O

PP	B-Simple_chemical
mesh	O
resulted	O
in	O
more	O
fibrous	B-Tissue
encapsulation	O
and	O
stiffness	O
than	O
PF	O
and	O
PS	O
prostheses	O
.	O

PP	B-Simple_chemical
also	O
resulted	O
in	O
less	O
connective	B-Tissue
tissue	I-Tissue
formation	O
and	O
foreign	O
-	O
body	O
reaction	O
than	O
PF	O
and	O
PS	O
prostheses	O
.	O

There	O
was	O
no	O
difference	O
in	O
fat	B-Tissue
encapsulation	O
,	O
necrosis	O
,	O
hemorrhage	O
,	O
or	O
exudate	B-Organism_substance
between	O
prostheses	O
.	O

Both	O
polyester	B-Simple_chemical
prostheses	O
(	O
PF	O
and	O
PS	O
)	O
have	O
better	O
tissue	B-Tissue
integration	O
than	O
the	O
PP	B-Simple_chemical
mesh	O
,	O
as	O
evidenced	O
by	O
the	O
higher	O
amount	O
of	O
connective	B-Tissue
tissue	I-Tissue
and	O
lower	O
extent	O
of	O
fibrous	B-Tissue
encapsulation	O
.	O

Integration	O
of	O
interferon	B-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
/	O
beta	B-Gene_or_gene_product
signalling	O
to	O
p53	B-Gene_or_gene_product
responses	O
in	O
tumour	B-Cancer
suppression	O
and	O
antiviral	B-Organism
defence	O
.	O

Swift	O
elimination	O
of	O
undesirable	O
cells	B-Cell
is	O
an	O
important	O
feature	O
in	O
tumour	B-Cancer
suppression	O
and	O
immunity	O
.	O

The	O
tumour	B-Cancer
suppressor	O
p53	B-Gene_or_gene_product
and	O
interferon	B-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
and	O
-	B-Gene_or_gene_product
beta	I-Gene_or_gene_product
(	O
IFN	B-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
/	O
beta	B-Gene_or_gene_product
)	O
are	O
essential	O
for	O
the	O
induction	O
of	O
apoptosis	O
in	O
cancerous	B-Cell
cells	I-Cell
and	O
in	O
antiviral	B-Organism
immune	O
responses	O
,	O
respectively	O
,	O
but	O
little	O
is	O
known	O
about	O
their	O
interrelationship	O
.	O

Here	O
we	O
show	O
that	O
transcription	O
of	O
the	O
p53	B-Gene_or_gene_product
gene	O
is	O
induced	O
by	O
IFN	B-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
/	O
beta	B-Gene_or_gene_product
,	O
accompanied	O
by	O
an	O
increase	O
in	O
p53	B-Gene_or_gene_product
protein	O
level	O
.	O

IFN	B-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
/	O
beta	B-Gene_or_gene_product
signalling	O
itself	O
does	O
not	O
activate	O
p53	B-Gene_or_gene_product
;	O
rather	O
,	O
it	O
contributes	O
to	O
boosting	O
p53	B-Gene_or_gene_product
responses	O
to	O
stress	O
signals	O
.	O

We	O
show	O
examples	O
in	O
which	O
p53	B-Gene_or_gene_product
gene	O
induction	O
by	O
IFN	B-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
/	O
beta	B-Gene_or_gene_product
contributes	O
to	O
tumour	B-Cancer
suppression	O
.	O

Furthermore	O
,	O
we	O
show	O
that	O
p53	B-Gene_or_gene_product
is	O
activated	O
in	O
virally	B-Organism
infected	O
cells	B-Cell
to	O
evoke	O
an	O
apoptotic	O
response	O
and	O
that	O
p53	B-Gene_or_gene_product
is	O
critical	O
for	O
antiviral	B-Organism
defence	O
of	O
the	O
host	O
.	O

Our	O
study	O
reveals	O
a	O
hitherto	O
unrecognized	O
link	O
between	O
p53	B-Gene_or_gene_product
and	O
IFN	B-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
/	O
beta	B-Gene_or_gene_product
in	O
tumour	B-Cancer
suppression	O
and	O
antiviral	B-Organism
immunity	O
,	O
which	O
may	O
have	O
therapeutic	O
implications	O
.	O

T140	B-Simple_chemical
analogs	I-Simple_chemical
as	O
CXCR4	B-Gene_or_gene_product
antagonists	O
identified	O
as	O
anti	O
-	O
metastatic	O
agents	O
in	O
the	O
treatment	O
of	O
breast	B-Cancer
cancer	I-Cancer
.	O

A	O
chemokine	O
receptor	O
,	O
CXCR4	B-Gene_or_gene_product
,	O
and	O
its	O
endogenous	O
ligand	O
,	O
stromal	B-Gene_or_gene_product
cell	I-Gene_or_gene_product
-	I-Gene_or_gene_product
derived	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
SDF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
,	O
have	O
been	O
recognized	O
to	O
be	O
involved	O
in	O
the	O
metastasis	O
of	O
several	O
types	O
of	O
cancers	B-Cancer
.	O

T140	B-Simple_chemical
analogs	I-Simple_chemical
are	O
peptidic	O
CXCR4	B-Gene_or_gene_product
antagonists	O
composed	O
of	O
14	O
amino	O
acid	O
residues	O
that	O
were	O
previously	O
developed	O
as	O
anti	O
-	O
HIV	B-Organism
agents	O
having	O
inhibitory	O
activity	O
against	O
HIV	B-Organism
-	O
entry	O
through	O
its	O
co	O
-	O
receptor	O
,	O
CXCR4	B-Gene_or_gene_product
.	O

Herein	O
,	O
we	O
report	O
that	O
these	O
compounds	O
effectively	O
inhibited	O
SDF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
induced	O
migration	O
of	O
human	B-Organism
breast	B-Cell
cancer	I-Cell
cells	I-Cell
(	O
MDA	B-Cell
-	I-Cell
MB	I-Cell
-	I-Cell
231	I-Cell
)	O
,	O
human	B-Organism
leukemia	B-Cell
T	I-Cell
cells	I-Cell
(	O
Sup	B-Cell
-	I-Cell
T1	I-Cell
)	O
and	O
human	B-Cell
umbilical	I-Cell
vein	I-Cell
endothelial	I-Cell
cells	I-Cell
at	O
concentrations	O
of	O
10	O
-	O
100	O
nM	O
in	O
vitro	O
.	O

Furthermore	O
,	O
slow	O
release	O
administration	O
by	O
subcutaneous	O
injection	O
using	O
an	O
Alzet	O
osmotic	O
pump	O
of	O
a	O
potent	O
and	O
bio	O
-	O
stable	O
T140	B-Simple_chemical
analog	I-Simple_chemical
,	O
4F	B-Simple_chemical
-	I-Simple_chemical
benzoyl	I-Simple_chemical
-	I-Simple_chemical
TN14003	I-Simple_chemical
,	O
gave	O
a	O
partial	O
,	O
but	O
statistically	O
significant	O
(	O
P	O
<	O
/	O
=	O
0	O
.	O
05	O
(	O
t	O
-	O
test	O
)	O
)	O
reduction	O
in	O
pulmonary	B-Organ
metastasis	O
of	O
MDA	B-Cell
-	I-Cell
MB	I-Cell
-	I-Cell
231	I-Cell
in	O
SCID	B-Organism
mice	I-Organism
,	O
even	O
though	O
no	O
attempt	O
was	O
made	O
to	O
inhibit	O
other	O
important	O
targets	O
such	O
as	O
CCR7	B-Gene_or_gene_product
.	O

These	O
results	O
suggest	O
that	O
T140	B-Simple_chemical
analogs	I-Simple_chemical
have	O
potential	O
use	O
for	O
cancer	B-Cancer
therapy	O
,	O
and	O
that	O
small	O
molecular	O
CXCR4	B-Gene_or_gene_product
antagonists	O
could	O
potentially	O
replace	O
neutralizing	O
antibodies	O
as	O
anti	O
-	O
metastatic	O
agents	O
for	O
breast	B-Cancer
cancer	I-Cancer
.	O

CCL16	B-Gene_or_gene_product
activates	O
an	O
angiogenic	O
program	O
in	O
vascular	B-Cell
endothelial	I-Cell
cells	I-Cell
.	O

Besides	O
regulating	O
leukocyte	B-Cell
trafficking	O
in	O
normal	O
and	O
injured	O
tissues	B-Tissue
,	O
several	O
chemokines	O
may	O
positively	O
or	O
negatively	O
regulate	O
angiogenesis	O
.	O

Here	O
we	O
report	O
that	O
CCL16	B-Gene_or_gene_product
activates	O
an	O
angiogenic	O
program	O
in	O
vascular	B-Cell
endothelial	I-Cell
cells	I-Cell
by	O
activating	O
CCR1	B-Gene_or_gene_product
.	O

CCL16	B-Gene_or_gene_product
induces	O
dose	O
-	O
dependent	O
random	O
and	O
directional	O
migration	O
of	O
endothelial	B-Cell
cells	I-Cell
isolated	O
from	O
large	O
vessels	B-Multi-tissue_structure
and	O
liver	B-Tissue
capillaries	I-Tissue
without	O
inducing	O
their	O
proliferation	O
.	O

It	O
also	O
promotes	O
endothelial	B-Cell
differentiation	O
into	O
capillary	B-Tissue
-	I-Tissue
like	I-Tissue
structures	I-Tissue
in	O
an	O
in	O
vitro	O
assay	O
and	O
is	O
angiogenic	O
in	O
the	O
chick	B-Organism
chorionallantoic	B-Multi-tissue_structure
membrane	I-Multi-tissue_structure
.	O

These	O
angiogenic	O
activities	O
are	O
neutralized	O
by	O
a	O
specific	O
antibody	O
against	O
CCL16	B-Gene_or_gene_product
.	O

The	O
direct	O
angiogenic	O
activity	O
of	O
CCL16	B-Gene_or_gene_product
is	O
further	O
amplified	O
by	O
its	O
ability	O
to	O
prime	O
endothelium	B-Tissue
to	O
a	O
mitogen	O
signal	O
induced	O
by	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
A	I-Gene_or_gene_product
and	O
to	O
raise	O
their	O
basal	O
production	O
of	O
CXCL8	B-Gene_or_gene_product
and	O
CCL2	B-Gene_or_gene_product
,	O
2	O
other	O
angiogenic	O
chemokines	O
.	O

BX471	B-Simple_chemical
(	O
R	B-Simple_chemical
-	I-Simple_chemical
N	I-Simple_chemical
-	I-Simple_chemical
[	I-Simple_chemical
5	I-Simple_chemical
-	I-Simple_chemical
chloro	I-Simple_chemical
-	I-Simple_chemical
2	I-Simple_chemical
-	I-Simple_chemical
[	I-Simple_chemical
2	I-Simple_chemical
-	I-Simple_chemical
[	I-Simple_chemical
4	I-Simple_chemical
(	I-Simple_chemical
4	I-Simple_chemical
-	I-Simple_chemical
fluorophenyl	I-Simple_chemical
)	I-Simple_chemical
methyl	I-Simple_chemical
]	I-Simple_chemical
-	I-Simple_chemical
2	I-Simple_chemical
-	I-Simple_chemical
methyl	I-Simple_chemical
-	I-Simple_chemical
1	I-Simple_chemical
-	I-Simple_chemical
piperazinyl	I-Simple_chemical
]	I-Simple_chemical
-	I-Simple_chemical
2	I-Simple_chemical
-	I-Simple_chemical
oxoethoxy	I-Simple_chemical
]	I-Simple_chemical
phenyl	I-Simple_chemical
]	I-Simple_chemical
urea	I-Simple_chemical
hydrochloric	I-Simple_chemical
acid	I-Simple_chemical
salt	I-Simple_chemical
)	O
,	O
a	O
CCR1	B-Gene_or_gene_product
antagonist	O
,	O
inhibits	O
angiogenic	O
properties	O
of	O
CCL16	B-Gene_or_gene_product
,	O
whereas	O
blocking	O
of	O
CCR8	B-Gene_or_gene_product
or	O
desensitizing	O
CCR2	B-Gene_or_gene_product
,	O
which	O
are	O
both	O
well	O
known	O
receptors	O
for	O
CCL16	B-Gene_or_gene_product
,	O
did	O
not	O
abolish	O
endothelial	B-Cell
activation	O
.	O

CCL16	B-Gene_or_gene_product
may	O
be	O
specifically	O
cross	O
-	O
linked	O
to	O
CCR1	B-Gene_or_gene_product
expressed	O
on	O
endothelial	B-Cell
cells	I-Cell
.	O

The	O
largely	O
restricted	O
CCL16	B-Gene_or_gene_product
expression	O
in	O
the	O
liver	B-Organ
suggests	O
that	O
this	O
chemokine	O
may	O
play	O
a	O
role	O
in	O
hepatic	B-Multi-tissue_structure
vascular	I-Multi-tissue_structure
formation	O
during	O
development	O
and	O
in	O
angiogenesis	O
associated	O
to	O
hepatic	B-Organ
diseases	O
.	O

Simvastatin	B-Simple_chemical
induces	O
apoptosis	O
of	O
B	B-Cell
-	I-Cell
CLL	I-Cell
cells	I-Cell
by	O
activation	O
of	O
mitochondrial	B-Cellular_component
caspase	B-Gene_or_gene_product
9	I-Gene_or_gene_product
.	O

BACKGROUND	O
AND	O
OBJECTIVES	O
:	O
Chronic	B-Cancer
lymphocytic	I-Cancer
leukemia	I-Cancer
(	O
CLL	B-Cancer
)	O
is	O
the	O
most	O
common	O
leukemia	B-Cancer
in	O
the	O
western	O
world	O
.	O

Despite	O
several	O
advances	O
in	O
therapeutic	O
options	O
,	O
the	O
disease	O
remains	O
incurable	O
.	O

Recently	O
,	O
it	O
was	O
repeatedly	O
demonstrated	O
that	O
statins	B-Simple_chemical
,	O
competitive	O
inhibitors	O
of	O
3	B-Gene_or_gene_product
-	I-Gene_or_gene_product
hydroxy	I-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
-	I-Gene_or_gene_product
methyl	I-Gene_or_gene_product
glutaryl	I-Gene_or_gene_product
coenzyme	I-Gene_or_gene_product
A	I-Gene_or_gene_product
(	I-Gene_or_gene_product
HMG	I-Gene_or_gene_product
-	I-Gene_or_gene_product
CoA	I-Gene_or_gene_product
)	I-Gene_or_gene_product
reductase	I-Gene_or_gene_product
,	O
have	O
antineoplastic	B-Cancer
effects	O
.	O

Therefore	O
we	O
aimed	O
to	O
study	O
the	O
effects	O
of	O
simvastatin	B-Simple_chemical
(	O
Sim	B-Simple_chemical
)	O
on	O
malignant	B-Cell
B	I-Cell
cells	I-Cell
derived	O
from	O
patients	B-Organism
with	O
CLL	B-Cancer
and	O
mechanisms	O
of	O
action	O
of	O
the	O
drug	O
.	O

METHODS	O
AND	O
RESULTS	O
:	O
Purified	O
B	B-Cell
-	I-Cell
CLL	I-Cell
cells	I-Cell
from	O
15	O
patients	B-Organism
were	O
cultured	O
either	O
alone	O
or	O
with	O
Sim	B-Simple_chemical
at	O
concentrations	O
of	O
10	O
,	O
50	O
,	O
and	O
100	O
microM	O
.	O

Viability	O
,	O
measured	O
by	O
the	O
activity	O
of	O
mitochondrial	B-Cellular_component
dehydrogenases	O
,	O
was	O
reduced	O
significantly	O
in	O
the	O
cells	B-Cell
treated	O
with	O
Sim	B-Simple_chemical
at	O
50	O
and	O
100	O
microM	O
for	O
24	O
hours	O
(	O
p	O
<	O
0	O
.	O
005	O
)	O
.	O

The	O
level	O
of	O
apoptosis	O
,	O
as	O
measured	O
by	O
annexin	B-Gene_or_gene_product
binding	O
to	O
exposed	O
phosphatidylserine	B-Amino_acid
moieties	O
,	O
increased	O
significantly	O
in	O
the	O
treated	O
cells	B-Cell
at	O
concentrations	O
higher	O
than	O
50	O
microM	O
for	O
24	O
hours	O
(	O
p	O
<	O
0	O
.	O
003	O
)	O
.	O

The	O
level	O
of	O
necrosis	O
,	O
as	O
measured	O
by	O
propidium	B-Simple_chemical
iodide	I-Simple_chemical
internalization	O
,	O
increased	O
significantly	O
after	O
24	O
hours	O
exposure	O
to	O
Sim	B-Simple_chemical
at	O
50	O
microM	O
(	O
p	O
<	O
0	O
.	O
01	O
)	O
.	O

The	O
apoptotic	O
cascade	O
was	O
studied	O
by	O
immunoblot	O
analysis	O
of	O
caspases	B-Gene_or_gene_product
following	O
Sim	B-Simple_chemical
treatment	O
.	O

These	O
showed	O
cleavage	O
of	O
caspases	B-Gene_or_gene_product
9	I-Gene_or_gene_product
,	O
8	B-Gene_or_gene_product
,	O
and	O
3	B-Gene_or_gene_product
.	O

Addition	O
of	O
the	O
caspase	B-Gene_or_gene_product
inhibitor	O
Z	B-Simple_chemical
-	I-Simple_chemical
VAD	I-Simple_chemical
.	I-Simple_chemical
fmk	I-Simple_chemical
inhibited	O
caspase	B-Gene_or_gene_product
8	I-Gene_or_gene_product
and	O
3	B-Gene_or_gene_product
significantly	O
but	O
did	O
not	O
affect	O
caspase	B-Gene_or_gene_product
9	I-Gene_or_gene_product
.	O

CONCLUSION	O
:	O
Exposure	O
of	O
clonal	B-Cell
B	I-Cell
lymphocytes	I-Cell
from	O
patients	B-Organism
with	O
CLL	B-Cancer
to	O
simvastatin	B-Simple_chemical
decreases	O
viability	O
significantly	O
by	O
the	O
induction	O
of	O
apoptosis	O
.	O

The	O
apoptosis	O
induced	O
by	O
Sim	B-Simple_chemical
is	O
probably	O
initiated	O
by	O
the	O
mitochondrial	B-Cellular_component
caspase	B-Gene_or_gene_product
9	I-Gene_or_gene_product
,	O
which	O
indirectly	O
leads	O
to	O
activation	O
of	O
caspase	B-Gene_or_gene_product
3	I-Gene_or_gene_product
and	O
8	B-Gene_or_gene_product
.	O

Autocrine	O
angiotensin	B-Gene_or_gene_product
system	O
regulation	O
of	O
bovine	B-Organism
aortic	B-Cell
endothelial	I-Cell
cell	I-Cell
migration	O
and	O
plasminogen	B-Gene_or_gene_product
activator	I-Gene_or_gene_product
involves	O
modulation	O
of	O
proto	O
-	O
oncogene	O
pp60c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
src	I-Gene_or_gene_product
expression	O
.	O

Rapid	O
endothelial	B-Cell
cell	I-Cell
migration	O
and	O
inhibition	O
of	O
thrombosis	O
are	O
critical	O
for	O
the	O
resolution	O
of	O
denudation	O
injuries	O
to	O
the	O
vessel	B-Multi-tissue_structure
wall	I-Multi-tissue_structure
.	O

Inhibition	O
of	O
the	O
endothelial	B-Cell
cell	I-Cell
autocrine	O
angiotensin	B-Gene_or_gene_product
system	O
,	O
with	O
either	O
the	O
angiotensin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
converting	I-Gene_or_gene_product
enzyme	I-Gene_or_gene_product
inhibitor	O
lisinopril	B-Simple_chemical
or	O
the	O
angiotensin	B-Gene_or_gene_product
II	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
antagonist	O
sar1	B-Gene_or_gene_product
,	O
ile8	B-Simple_chemical
-	I-Simple_chemical
angiotensin	I-Simple_chemical
II	I-Simple_chemical
,	I-Simple_chemical
leads	O
to	O
increased	O
endothelial	B-Cell
cell	I-Cell
migration	O
and	O
urokinase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
like	I-Gene_or_gene_product
plasminogen	I-Gene_or_gene_product
activator	I-Gene_or_gene_product
(	O
u	B-Gene_or_gene_product
-	I-Gene_or_gene_product
PA	I-Gene_or_gene_product
)	O
activity	O
(	O
Bell	O
,	O
L	O
.	O
,	O
and	O
J	O
.	O
A	O
.	O
Madri	O
.	O
1990	O
.	O
Am	O
.	O
J	O
.	O
Pathol	O
.	O
137	O
:	O
7	O
-	O
12	O
)	O
.	O

Inhibition	O
of	O
the	O
autocrine	O
angiotensin	B-Gene_or_gene_product
system	O
with	O
the	O
converting	O
-	O
enzyme	O
inhibitor	O
or	O
the	O
receptor	O
antagonist	O
also	O
leads	O
to	O
increased	O
expression	O
of	O
the	O
proto	O
-	O
oncogene	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
src	I-Gene_or_gene_product
:	O
pp60c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
src	I-Gene_or_gene_product
mRNA	O
increased	O
7	O
-	O
11	O
-	O
fold	O
,	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
src	I-Gene_or_gene_product
protein	O
3	O
-	O
fold	O
,	O
and	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
src	I-Gene_or_gene_product
kinase	O
activity	O
2	O
-	O
3	O
-	O
fold	O
.	O

Endothelial	B-Cell
cell	I-Cell
expression	O
of	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
src	I-Gene_or_gene_product
was	O
constitutively	O
elevated	O
after	O
stable	O
infection	O
with	O
a	O
retroviral	O
vector	O
containing	O
the	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
src	I-Gene_or_gene_product
coding	O
sequence	O
.	O

Constitutively	O
increased	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
src	I-Gene_or_gene_product
kinase	O
activity	O
reconstituted	O
the	O
increases	O
in	O
migration	O
and	O
u	B-Gene_or_gene_product
-	I-Gene_or_gene_product
PA	I-Gene_or_gene_product
observed	O
with	O
angiotensin	B-Gene_or_gene_product
system	O
interruption	O
.	O

Antisera	O
to	O
bovine	B-Organism
u	B-Gene_or_gene_product
-	I-Gene_or_gene_product
PA	I-Gene_or_gene_product
blocked	O
the	O
increase	O
in	O
migration	O
associated	O
with	O
increased	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
src	I-Gene_or_gene_product
expression	O
.	O

These	O
data	O
suggest	O
that	O
increases	O
in	O
endothelial	B-Cell
cell	I-Cell
migration	O
and	O
plasminogen	B-Gene_or_gene_product
activator	I-Gene_or_gene_product
after	O
angiotensin	B-Gene_or_gene_product
system	O
inhibition	O
are	O
at	O
least	O
partially	O
pp60c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
src	I-Gene_or_gene_product
mediated	O
.	O

Elevated	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
src	I-Gene_or_gene_product
expression	O
with	O
angiotensin	B-Gene_or_gene_product
system	O
inhibition	O
may	O
act	O
to	O
enhance	O
intimal	B-Pathological_formation
wound	I-Pathological_formation
closure	O
and	O
to	O
reduce	O
luminal	B-Immaterial_anatomical_entity
thrombogenicity	O
in	O
vivo	O
.	O

Cell	B-Cell
type	O
-	O
specific	O
regulation	O
of	O
angiogenic	O
growth	O
factor	O
gene	O
expression	O
and	O
induction	O
of	O
angiogenesis	O
in	O
nonischemic	B-Tissue
tissue	I-Tissue
by	O
a	O
constitutively	O
active	O
form	O
of	O
hypoxia	B-Gene_or_gene_product
-	I-Gene_or_gene_product
inducible	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

Understanding	O
molecular	O
mechanisms	O
regulating	O
angiogenesis	O
may	O
lead	O
to	O
novel	O
therapies	O
for	O
ischemic	O
disorders	O
.	O

Hypoxia	B-Gene_or_gene_product
-	I-Gene_or_gene_product
inducible	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
activates	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
gene	O
expression	O
in	O
hypoxic	B-Tissue
/	O
ischemic	B-Tissue
tissue	I-Tissue
.	O

In	O
this	O
study	O
we	O
demonstrate	O
that	O
exposure	O
of	O
primary	O
cultures	O
of	O
cardiac	B-Cell
and	O
vascular	B-Cell
cells	I-Cell
to	O
hypoxia	O
or	O
AdCA5	B-Organism
,	O
an	O
adenovirus	O
encoding	O
a	O
constitutively	O
active	O
form	O
of	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
,	O
modulates	O
the	O
expression	O
of	O
genes	O
encoding	O
the	O
angiogenic	O
factors	O
angiopoietin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
ANGPT1	B-Gene_or_gene_product
)	O
,	O
ANGPT2	B-Gene_or_gene_product
,	O
placental	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
,	O
and	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
-	I-Gene_or_gene_product
B	I-Gene_or_gene_product
.	O

Loss	O
-	O
of	O
-	O
function	O
effects	O
were	O
also	O
observed	O
in	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
-	O
null	O
embryonic	O
stem	O
cells	O
.	O

Depending	O
on	O
the	O
cell	B-Cell
type	O
,	O
expression	O
of	O
ANGPT1	B-Gene_or_gene_product
and	O
ANGPT2	B-Gene_or_gene_product
was	O
either	O
activated	O
or	O
repressed	O
in	O
response	O
to	O
hypoxia	O
or	O
AdCA5	B-Organism
.	O

In	O
all	O
cases	O
,	O
there	O
was	O
complete	O
concordance	O
between	O
the	O
effects	O
of	O
hypoxia	O
and	O
AdCA5	B-Organism
.	O

Injection	O
of	O
AdCA5	B-Organism
into	O
mouse	B-Organism
eyes	B-Organ
induced	O
neovascularization	O
in	O
multiple	O
capillary	B-Multi-tissue_structure
beds	I-Multi-tissue_structure
,	O
including	O
those	O
not	O
responsive	O
to	O
VEGF	B-Gene_or_gene_product
alone	O
.	O

Analysis	O
of	O
gene	O
expression	O
revealed	O
increased	O
expression	O
of	O
ANGPT1	B-Gene_or_gene_product
,	O
ANGPT2	B-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
-	I-Gene_or_gene_product
B	I-Gene_or_gene_product
,	O
placental	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
,	O
and	O
VEGF	B-Gene_or_gene_product
mRNA	O
in	O
AdCA5	B-Organism
-	O
injected	O
eyes	B-Organ
.	O

These	O
results	O
indicate	O
that	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
functions	O
as	O
a	O
master	O
regulator	O
of	O
angiogenesis	O
by	O
controlling	O
the	O
expression	O
of	O
multiple	O
angiogenic	O
growth	O
factors	O
and	O
that	O
adenovirus	O
-	O
mediated	O
expression	O
of	O
a	O
constitutively	O
active	O
form	O
of	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
is	O
sufficient	O
to	O
induce	O
angiogenesis	O
in	O
nonischemic	B-Tissue
tissue	I-Tissue
of	O
an	O
adult	O
animal	O
.	O

Differential	O
regulation	O
of	O
in	O
vivo	O
angiogenesis	O
by	O
angiotensin	B-Gene_or_gene_product
II	I-Gene_or_gene_product
receptors	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
,	O
a	O
key	O
regulator	O
of	O
blood	B-Organism_substance
pressure	O
and	O
body	B-Organism_substance
fluid	I-Organism_substance
homeostasis	O
,	O
exerts	O
mitogenic	O
effects	O
on	O
endothelial	B-Cell
cells	I-Cell
.	O

We	O
therefore	O
hypothesized	O
that	O
ANG	B-Gene_or_gene_product
II	I-Gene_or_gene_product
could	O
be	O
a	O
mediator	O
between	O
homeostatic	O
changes	O
within	O
the	O
vascular	B-Multi-tissue_structure
perfusion	I-Multi-tissue_structure
bed	I-Multi-tissue_structure
and	O
growth	O
factor	O
-	O
driven	O
angiogenesis	O
.	O

In	O
the	O
present	O
study	O
,	O
we	O
applied	O
the	O
alginate	B-Simple_chemical
implant	O
angiogenesis	O
model	O
in	O
mice	B-Organism
with	O
normal	O
ANG	B-Gene_or_gene_product
II	I-Gene_or_gene_product
levels	O
,	O
elevated	O
ANG	B-Gene_or_gene_product
II	I-Gene_or_gene_product
levels	O
by	O
transgenic	O
overexpression	O
of	O
angiotensinogen	B-Gene_or_gene_product
(	O
AOGEN	B-Gene_or_gene_product
)	O
,	O
or	O
in	O
AT2	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
-	O
deficient	O
mice	B-Organism
.	O

We	O
demonstrate	O
that	O
a	O
decrease	O
in	O
the	O
amount	O
of	O
circulating	O
ANG	B-Gene_or_gene_product
II	I-Gene_or_gene_product
by	O
the	O
angiotensin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
converting	I-Gene_or_gene_product
enzyme	I-Gene_or_gene_product
(	O
ACE	B-Gene_or_gene_product
)	O
inhibitor	O
enalapril	B-Simple_chemical
or	O
the	O
AT1	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
antagonist	O
losartan	B-Simple_chemical
induced	O
a	O
stimulation	O
of	O
in	O
vivo	O
angiogenesis	O
implying	O
an	O
inhibitory	O
function	O
of	O
ANG	B-Gene_or_gene_product
II	I-Gene_or_gene_product
through	O
the	O
AT1	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
.	O

However	O
,	O
the	O
strong	O
increase	O
of	O
angiogenesis	O
in	O
AOGEN	B-Gene_or_gene_product
-	O
transgenic	O
mice	B-Organism
compared	O
with	O
mice	B-Organism
with	O
normal	O
ANG	B-Gene_or_gene_product
II	I-Gene_or_gene_product
levels	O
suggests	O
additional	O
stimulatory	O
activity	O
.	O

We	O
showed	O
that	O
the	O
ANG	B-Gene_or_gene_product
II	I-Gene_or_gene_product
-	O
induced	O
stimulation	O
of	O
angiogenesis	O
is	O
linked	O
to	O
the	O
AT2	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
as	O
an	O
impaired	O
induction	O
of	O
angiogenesis	O
was	O
obtained	O
in	O
AT2	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
knockout	O
mice	O
.	O

These	O
findings	O
provide	O
the	O
first	O
evidence	O
that	O
the	O
AT2	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
mediates	O
a	O
stimulation	O
of	O
in	O
vivo	O
angiogenesis	O
and	O
indicate	O
that	O
ANG	B-Gene_or_gene_product
II	I-Gene_or_gene_product
is	O
a	O
humoral	O
regulator	O
of	O
peripheral	O
angiogenesis	O
involving	O
two	O
receptor	O
subtypes	O
with	O
opposing	O
actions	O
.	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
receptor	I-Gene_or_gene_product
enhances	O
invasion	O
and	O
induces	O
resistance	O
to	O
apoptosis	O
of	O
colon	B-Cell
cancer	I-Cell
cells	I-Cell
through	O
the	O
Akt	B-Gene_or_gene_product
/	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
x	I-Gene_or_gene_product
(	I-Gene_or_gene_product
L	I-Gene_or_gene_product
)	I-Gene_or_gene_product
pathway	O
.	O

Colon	B-Cancer
cancer	I-Cancer
overexpresses	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
(	O
IGF1	B-Gene_or_gene_product
)	O
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
receptor	I-Gene_or_gene_product
(	O
IGF1	B-Gene_or_gene_product
-	I-Gene_or_gene_product
R	I-Gene_or_gene_product
)	O
,	O
as	O
compared	O
with	O
normal	O
or	O
adenomatous	B-Multi-tissue_structure
mucosa	I-Multi-tissue_structure
,	O
and	O
it	O
has	O
been	O
postulated	O
that	O
colorectal	B-Cell
cancer	I-Cell
cells	I-Cell
depend	O
on	O
the	O
IGF1	B-Gene_or_gene_product
/	O
IGF1	B-Gene_or_gene_product
-	I-Gene_or_gene_product
R	I-Gene_or_gene_product
pathway	O
for	O
their	O
growth	O
and	O
progression	O
.	O

In	O
this	O
study	O
,	O
using	O
the	O
human	B-Organism
colon	B-Cell
cancer	I-Cell
cell	I-Cell
line	I-Cell
HCT116	I-Cell
,	O
we	O
find	O
that	O
established	O
HCT116	O
/	O
IGF1	B-Gene_or_gene_product
-	I-Gene_or_gene_product
R	I-Gene_or_gene_product
transfectants	O
exhibit	O
a	O
more	O
aggressive	O
transformed	O
phenotype	O
than	O
the	O
parental	B-Cell
cell	I-Cell
line	I-Cell
,	O
as	O
demonstrated	O
by	O
their	O
higher	O
proliferation	O
rate	O
in	O
response	O
to	O
IGF1	B-Gene_or_gene_product
,	O
higher	O
degree	O
of	O
anchorage	O
-	O
independent	O
growth	O
,	O
resistance	O
to	O
serum	B-Organism_substance
deprivation	O
-	O
induced	O
apoptosis	O
,	O
and	O
higher	O
migratory	O
capability	O
in	O
a	O
monolayer	B-Cell
"	O
wounding	O
assay	O
.	O
"	O
When	O
injected	O
into	O
nude	B-Organism
mice	I-Organism
,	O
HCT116	O
/	O
IGF1	B-Gene_or_gene_product
-	I-Gene_or_gene_product
R	I-Gene_or_gene_product
transfectants	O
were	O
highly	O
invasive	O
and	O
produced	O
distant	O
metastases	B-Cancer
,	O
whereas	O
the	O
parental	B-Cell
cell	I-Cell
did	O
not	O
.	O

Moreover	O
,	O
the	O
overexpression	O
of	O
IGF1	B-Gene_or_gene_product
-	I-Gene_or_gene_product
R	I-Gene_or_gene_product
in	O
these	O
cells	B-Cell
was	O
associated	O
with	O
IGF1	B-Gene_or_gene_product
-	I-Gene_or_gene_product
R	I-Gene_or_gene_product
-	O
induced	O
activation	O
of	O
Akt	B-Gene_or_gene_product
and	O
up	O
-	O
regulation	O
of	O
the	O
antiapoptotic	O
protein	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
x	I-Gene_or_gene_product
(	I-Gene_or_gene_product
L	I-Gene_or_gene_product
)	I-Gene_or_gene_product
.	O

We	O
also	O
show	O
that	O
Akt	B-Gene_or_gene_product
pathway	O
mediates	O
IGF1	B-Gene_or_gene_product
-	I-Gene_or_gene_product
R	I-Gene_or_gene_product
-	O
induced	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
x	I-Gene_or_gene_product
(	I-Gene_or_gene_product
L	I-Gene_or_gene_product
)	I-Gene_or_gene_product
expression	O
at	O
transcriptional	O
level	O
.	O

Our	O
data	O
demonstrate	O
,	O
for	O
the	O
first	O
time	O
,	O
that	O
IGF1	B-Gene_or_gene_product
-	I-Gene_or_gene_product
R	I-Gene_or_gene_product
/	O
Akt	B-Gene_or_gene_product
/	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
x	I-Gene_or_gene_product
(	I-Gene_or_gene_product
L	I-Gene_or_gene_product
)	I-Gene_or_gene_product
pathway	O
may	O
contribute	O
to	O
a	O
more	O
aggressive	O
malignant	O
phenotype	O
,	O
in	O
a	O
subset	O
of	O
colorectal	B-Cancer
cancers	I-Cancer
.	O

Expression	O
of	O
EphA2	B-Gene_or_gene_product
and	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
in	O
colorectal	B-Cancer
cancer	I-Cancer
:	O
correlation	O
with	O
cancer	B-Cancer
metastasis	O
.	O

Recently	O
,	O
overexpression	O
of	O
EphA2	B-Gene_or_gene_product
,	O
a	O
member	O
of	O
the	O
Eph	B-Gene_or_gene_product
family	O
of	O
receptor	O
tyrosine	O
kinases	O
,	O
has	O
been	O
reported	O
in	O
several	O
cancers	B-Cancer
.	O

Reduced	O
expression	O
of	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
,	O
an	O
intercellular	B-Immaterial_anatomical_entity
adhesion	O
molecule	O
of	O
epithelial	B-Cell
cells	I-Cell
,	O
has	O
been	O
reported	O
to	O
be	O
associated	O
with	O
aggressive	O
clinicopathological	O
phenotypes	O
in	O
various	O
cancers	B-Cancer
.	O

In	O
epithelial	B-Cell
cells	I-Cell
,	O
EphA2	B-Gene_or_gene_product
and	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
co	O
-	O
localize	O
to	O
sites	O
of	O
cell	B-Cellular_component
-	I-Cellular_component
cell	I-Cellular_component
contact	I-Cellular_component
,	O
and	O
it	O
has	O
been	O
shown	O
that	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
regulates	O
EphA2	B-Gene_or_gene_product
.	O

This	O
study	O
aimed	O
to	O
clarify	O
the	O
relationship	O
between	O
the	O
expression	O
of	O
the	O
EphA2	B-Gene_or_gene_product
and	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
proteins	O
and	O
clinicopathological	O
characteristics	O
,	O
with	O
reference	O
to	O
the	O
expression	O
levels	O
of	O
both	O
EphA2	B-Gene_or_gene_product
and	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
,	O
in	O
patients	B-Organism
with	O
colorectal	B-Cancer
cancer	I-Cancer
.	O

We	O
performed	O
immunohistochemical	O
staining	O
of	O
EphA2	B-Gene_or_gene_product
and	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
with	O
EphA2	B-Gene_or_gene_product
and	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
monoclonal	O
antibodies	O
in	O
samples	O
from	O
194	O
primary	B-Cancer
lesions	I-Cancer
of	O
colorectal	B-Cancer
cancer	I-Cancer
.	O

The	O
expression	O
level	O
of	O
EphA2	B-Gene_or_gene_product
had	O
a	O
statistically	O
significant	O
relationship	O
with	O
liver	B-Organ
metastasis	O
,	O
lymphatic	B-Multi-tissue_structure
vessel	I-Multi-tissue_structure
invasion	O
and	O
clinical	O
stage	O
(	O
p	O
=	O
0	O
.	O
0477	O
,	O
0	O
.	O
0316	O
and	O
0	O
.	O
0467	O
,	O
respectively	O
)	O
.	O

In	O
addition	O
,	O
the	O
positivity	O
rate	O
of	O
EphA2	B-Gene_or_gene_product
was	O
significantly	O
higher	O
in	O
primary	B-Cancer
lesions	I-Cancer
with	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
than	O
in	O
those	O
without	O
metastasis	O
(	O
p	O
=	O
0	O
.	O
0014	O
)	O
.	O

However	O
,	O
the	O
expression	O
level	O
of	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
had	O
an	O
inverse	O
relationship	O
with	O
both	O
differentiation	O
level	O
of	O
the	O
tumor	B-Cancer
and	O
lymphatic	B-Multi-tissue_structure
vessel	I-Multi-tissue_structure
invasion	O
(	O
p	O
=	O
0	O
.	O
0430	O
and	O
0	O
.	O
0320	O
,	O
respectively	O
)	O
.	O

Furthermore	O
,	O
a	O
significant	O
relationship	O
between	O
the	O
expression	O
of	O
EphA2	B-Gene_or_gene_product
and	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
was	O
observed	O
.	O

In	O
conclusion	O
,	O
our	O
study	O
revealed	O
that	O
the	O
overexpression	O
of	O
EphA2	B-Gene_or_gene_product
protein	O
in	O
colorectal	B-Tissue
carcinoma	I-Tissue
tissue	I-Tissue
correlates	O
closely	O
with	O
cancer	B-Cancer
progression	O
and	O
hematogenous	O
and	O
lymphogenous	O
metastasis	O
,	O
suggesting	O
that	O
both	O
EphA2	B-Gene_or_gene_product
and	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
may	O
play	O
an	O
important	O
role	O
in	O
tumor	B-Cancer
metastasis	O
in	O
colorectal	B-Cancer
cancer	I-Cancer
.	O

Nuclear	B-Cellular_component
translocation	O
of	O
a	O
clusterin	B-Gene_or_gene_product
isoform	O
is	O
associated	O
with	O
induction	O
of	O
anoikis	O
in	O
SV40	B-Organism
-	O
immortalized	O
human	B-Organism
prostate	B-Cell
epithelial	I-Cell
cells	I-Cell
.	O

Clusterin	B-Gene_or_gene_product
gene	O
expression	O
is	O
potently	O
induced	O
in	O
experimental	O
models	O
in	O
which	O
apoptosis	O
is	O
activated	O
,	O
such	O
as	O
rat	B-Organism
prostate	B-Organ
involution	O
following	O
castration	O
.	O

Nevertheless	O
,	O
its	O
precise	O
physiological	O
role	O
has	O
not	O
yet	O
been	O
established	O
,	O
and	O
both	O
anti	O
-	O
apoptotic	O
and	O
pro	O
-	O
apoptotic	O
functions	O
have	O
been	O
suggested	O
for	O
this	O
gene	O
.	O

Clusterin	B-Gene_or_gene_product
expression	O
level	O
depends	O
on	O
cell	B-Cell
proliferation	O
state	O
,	O
and	O
we	O
recently	O
showed	O
that	O
its	O
over	O
-	O
expression	O
inhibited	O
cell	B-Cell
cycle	O
progression	O
of	O
SV40	B-Organism
-	O
immortalized	O
human	B-Organism
prostate	B-Cell
epithelial	I-Cell
cells	I-Cell
PNT2	I-Cell
and	O
PNT1a	B-Cell
.	O

Here	O
we	O
studied	O
clusterin	B-Gene_or_gene_product
expression	O
in	O
PNT1a	B-Cell
cells	I-Cell
subjected	O
to	O
serum	B-Organism_substance
-	O
starvation	O
with	O
the	O
aim	O
of	O
defining	O
clusterin	B-Gene_or_gene_product
early	O
molecular	O
changes	O
following	O
apoptosis	O
induction	O
.	O

Under	O
serum	B-Organism_substance
-	O
starvation	O
conditions	O
,	O
decreased	O
growth	O
rate	O
,	O
slow	O
rounding	O
-	O
up	O
of	O
cells	B-Cell
,	O
cell	B-Cell
detachment	O
,	O
and	O
formation	O
of	O
apoptotic	B-Cellular_component
bodies	I-Cellular_component
indicative	O
of	O
anoikis	O
(	O
detachment	O
-	O
induced	O
apoptosis	O
)	O
were	O
preceded	O
by	O
significant	O
downregulation	O
of	O
70	O
kDa	O
clusterin	B-Gene_or_gene_product
precursor	O
and	O
upregulation	O
of	O
45	O
-	O
40	O
kDa	O
isoforms	O
.	O

On	O
the	O
8th	O
day	O
of	O
serum	B-Organism_substance
-	O
free	O
culturing	O
,	O
only	O
the	O
higher	O
molecular	O
weight	O
protein	O
-	O
band	O
of	O
about	O
45	O
kDa	O
was	O
clearly	O
induced	O
and	O
accumulated	O
in	O
detached	O
cells	B-Cell
and	O
apoptotic	B-Cellular_component
bodies	I-Cellular_component
in	O
which	O
PARP	B-Gene_or_gene_product
was	O
activated	O
.	O

Anoikis	O
was	O
preceded	O
by	O
induction	O
and	O
transloction	O
of	O
a	O
45	O
-	O
kDa	O
clusterin	B-Gene_or_gene_product
isoform	O
to	O
the	O
nucleus	B-Cellular_component
.	O

Thus	O
,	O
nuclear	B-Cellular_component
targeting	O
of	O
a	O
specific	O
45	O
-	O
kDa	O
isoform	O
of	O
clusterin	B-Gene_or_gene_product
appeared	O
to	O
be	O
an	O
early	O
and	O
specific	O
molecular	O
signal	O
triggering	O
anoikis	O
-	O
death	O
.	O

Considering	O
also	O
that	O
clusterin	B-Gene_or_gene_product
is	O
downregulated	O
during	O
prostate	B-Cancer
cancer	I-Cancer
onset	O
and	O
progression	O
,	O
and	O
that	O
its	O
upregulation	O
has	O
inhibited	O
DNA	B-Cellular_component
synthesis	O
and	O
cell	B-Cell
cycle	O
progression	O
of	O
immortalized	O
human	B-Organism
prostate	B-Cell
epithelial	I-Cell
cells	I-Cell
,	O
we	O
suggest	O
that	O
clusterin	B-Gene_or_gene_product
might	O
be	O
a	O
new	O
anti	O
-	O
oncogene	O
in	O
the	O
prostate	B-Organ
.	O

Levels	O
of	O
expression	O
of	O
CYR61	B-Gene_or_gene_product
and	O
CTGF	B-Gene_or_gene_product
are	O
prognostic	O
for	O
tumor	B-Cancer
progression	O
and	O
survival	O
of	O
individuals	B-Organism
with	O
gliomas	B-Cancer
.	O

The	O
biological	O
properties	O
of	O
CCN	B-Gene_or_gene_product
proteins	O
include	O
stimulation	O
of	O
cell	B-Cell
proliferation	O
,	O
migration	O
,	O
and	O
adhesion	O
,	O
as	O
well	O
as	O
angiogenesis	O
and	O
tumorigenesis	O
.	O

We	O
quantified	O
CYR61	B-Gene_or_gene_product
,	O
CTGF	B-Gene_or_gene_product
,	O
WISP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
and	O
NOV	B-Gene_or_gene_product
mRNA	O
expression	O
levels	O
in	O
samples	B-Cancer
from	O
sixty	O
-	O
six	O
primary	O
gliomas	B-Cancer
and	O
five	O
normal	O
brain	B-Multi-tissue_structure
samples	I-Multi-tissue_structure
using	O
quantitative	O
real	O
-	O
time	O
PCR	O
assay	O
.	O

Statistical	O
analysis	O
was	O
performed	O
to	O
explore	O
the	O
links	O
between	O
expression	O
of	O
the	O
CCN	B-Gene_or_gene_product
genes	O
and	O
clinical	O
and	O
pathological	O
parameters	O
.	O

Overexpression	O
of	O
CYR61	B-Gene_or_gene_product
,	O
CTGF	B-Gene_or_gene_product
,	O
WISP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
and	O
NOV	B-Gene_or_gene_product
occurred	O
in	O
48	O
%	O
(	O
32	O
of	O
66	O
)	O
,	O
58	O
%	O
(	O
38	O
of	O
66	O
)	O
,	O
36	O
%	O
(	O
24	O
of	O
66	O
)	O
,	O
and	O
15	O
%	O
(	O
10	O
of	O
66	O
)	O
of	O
primary	O
gliomas	B-Cancer
,	O
respectively	O
.	O

Interestingly	O
,	O
significant	O
associations	O
were	O
found	O
between	O
CYR61	B-Gene_or_gene_product
expression	O
versus	O
tumor	B-Cancer
grade	O
,	O
pathology	O
,	O
gender	O
,	O
and	O
age	O
at	O
diagnosis	O
.	O

Also	O
,	O
a	O
significant	O
correlation	O
existed	O
between	O
CTGF	B-Gene_or_gene_product
mRNA	O
levels	O
versus	O
tumor	B-Cancer
grade	O
,	O
gender	O
,	O
and	O
pathology	O
.	O

In	O
contrast	O
to	O
CYR61	B-Gene_or_gene_product
and	O
CTGF	B-Gene_or_gene_product
,	O
no	O
significant	O
association	O
was	O
found	O
between	O
expression	O
of	O
either	O
WISP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
or	O
NOV	B-Gene_or_gene_product
versus	O
any	O
of	O
the	O
pathological	O
features	O
.	O

Furthermore	O
,	O
Cox	O
regression	O
analysis	O
showed	O
that	O
CYR61	B-Gene_or_gene_product
and	O
CTGF	B-Gene_or_gene_product
expression	O
had	O
a	O
significant	O
correlation	O
with	O
patient	B-Organism
survival	O
.	O

These	O
results	O
suggest	O
that	O
CYR61	B-Gene_or_gene_product
and	O
CTGF	B-Gene_or_gene_product
may	O
play	O
a	O
role	O
in	O
the	O
progression	O
of	O
gliomas	B-Cancer
;	O
their	O
levels	O
at	O
diagnosis	O
may	O
have	O
prognostic	O
significance	O
;	O
and	O
these	O
proteins	O
might	O
serve	O
as	O
valuable	O
targets	O
for	O
therapeutic	O
intervention	O
.	O

Targeting	O
wide	O
-	O
range	O
oncogenic	O
transformation	O
via	O
PU24FCl	B-Simple_chemical
,	O
a	O
specific	O
inhibitor	O
of	O
tumor	B-Cancer
Hsp90	B-Gene_or_gene_product
.	O

Agents	O
that	O
inhibit	O
Hsp90	B-Gene_or_gene_product
function	O
hold	O
significant	O
promise	O
in	O
cancer	B-Cancer
therapy	O
.	O

Here	O
we	O
present	O
PU24FCl	B-Simple_chemical
,	O
a	O
representative	O
of	O
the	O
first	O
class	O
of	O
designed	O
Hsp90	B-Gene_or_gene_product
inhibitors	O
.	O

By	O
specifically	O
and	O
potently	O
inhibiting	O
tumor	B-Cancer
Hsp90	B-Gene_or_gene_product
,	O
PU24FCl	B-Simple_chemical
exhibits	O
wide	O
-	O
ranging	O
anti	O
-	O
cancer	B-Cancer
activities	O
that	O
occur	O
at	O
similar	O
doses	O
in	O
all	O
tested	O
tumor	B-Cancer
types	O
.	O

Normal	B-Cell
cells	I-Cell
are	O
10	O
-	O
to	O
50	O
-	O
fold	O
more	O
resistant	O
to	O
these	O
effects	O
.	O

Its	O
Hsp90	B-Gene_or_gene_product
inhibition	O
results	O
in	O
multiple	O
anti	O
-	O
tumor	B-Cancer
-	O
specific	O
effects	O
,	O
such	O
as	O
degradation	O
of	O
Hsp90	B-Gene_or_gene_product
-	O
client	O
proteins	O
involved	O
in	O
cell	B-Cell
growth	O
,	O
survival	O
,	O
and	O
specific	O
transformation	O
,	O
inhibition	O
of	O
cancer	B-Cell
cell	I-Cell
growth	O
,	O
delay	O
of	O
cell	B-Cell
cycle	O
progression	O
,	O
induction	O
of	O
morphological	O
and	O
functional	O
changes	O
,	O
and	O
apoptosis	O
.	O

In	O
concordance	O
with	O
its	O
higher	O
affinity	O
for	O
tumor	B-Cancer
Hsp90	B-Gene_or_gene_product
,	O
in	O
vivo	O
PU24FCl	B-Simple_chemical
accumulates	O
in	O
tumors	B-Cancer
while	O
being	O
rapidly	O
cleared	O
from	O
normal	B-Tissue
tissue	I-Tissue
.	O

Concentrations	O
achieved	O
in	O
vivo	O
in	O
tumors	B-Cancer
lead	O
to	O
single	O
-	O
agent	O
anti	O
-	O
tumor	B-Cancer
activity	O
at	O
non	O
-	O
toxic	O
doses	O
.	O

Differential	O
proteomic	O
analysis	O
of	O
human	B-Organism
hepatocellular	B-Cell
carcinoma	I-Cell
cell	I-Cell
line	I-Cell
metastasis	O
-	O
associated	O
proteins	O
.	O

PURPOSE	O
:	O
The	O
comparative	O
study	O
of	O
differentially	O
expression	O
of	O
protein	O
profiles	O
of	O
hepatocellular	B-Cell
carcinoma	I-Cell
cell	I-Cell
lines	I-Cell
with	O
various	O
metastasic	O
potential	O
and	O
screening	O
key	O
molecules	O
related	O
to	O
hepatocellular	B-Cancer
carcinoma	I-Cancer
metastasis	O
and	O
recurrence	O
.	O

METHODS	O
:	O
Using	O
two	O
-	O
dimensional	O
electrophoresis	O
and	O
liquid	O
chromatography	O
-	O
electrospray	O
ionization	O
-	O
tandem	O
mass	O
spectrometry	O
(	O
LC	O
-	O
ESI	O
-	O
MS	O
/	O
MS	O
)	O
,	O
we	O
analyzed	O
differentially	O
displayed	O
proteomics	O
of	O
human	B-Organism
hepatocellular	B-Cell
carcinoma	I-Cell
cell	I-Cell
lines	I-Cell
Hep3B	I-Cell
,	O
MHCC97L	B-Cell
,	O
MHCC97H	B-Cell
with	O
different	O
metastasic	O
potential	O
.	O

RESULTS	O
:	O
Approximate	O
1	O
,	O
000	O
protein	O
spots	O
were	O
detected	O
on	O
silver	B-Simple_chemical
-	O
stained	O
gel	O
by	O
ImageMaster	O
(	O
977	O
+	O
/	O
-	O
113	O
spots	O
in	O
Hep3B	B-Cell
,	O
1092	O
+	O
/	O
-	O
40	O
in	O
MHCC97L	B-Cell
,	O
and	O
889	O
+	O
/	O
-	O
14	O
in	O
MHCC97H	B-Cell
)	O
.	O

Fifty	O
distinct	O
different	O
protein	O
spots	O
were	O
analyzed	O
with	O
online	O
LC	O
-	O
ESI	O
-	O
MS	O
/	O
MS	O
.	O

Only	O
26	O
protein	O
spots	O
had	O
a	O
positive	O
result	O
,	O
including	O
annexin1	B-Gene_or_gene_product
,	O
S100A4	B-Gene_or_gene_product
,	O
and	O
so	O
on	O
.	O

In	O
comparison	O
with	O
nonmetastasis	O
Hep3B	B-Cell
cell	I-Cell
lines	I-Cell
,	O
there	O
were	O
16	O
proteins	O
overexpressed	O
in	O
MHCC97H	B-Cell
and	O
MHCC97L	B-Cell
,	O
10	O
proteins	O
underexpressed	O
in	O
MHCC97H	B-Cell
and	O
MHCC97L	B-Cell
.	O

Applying	O
cell	B-Cell
immunohistochemistry	O
and	O
RT	O
-	O
PCR	O
,	O
we	O
further	O
validated	O
two	O
interesting	O
and	O
different	O
proteins	O
,	O
annexin1	B-Gene_or_gene_product
and	O
S100A4	B-Gene_or_gene_product
.	O

CONCLUSION	O
:	O
The	O
protein	O
profile	O
of	O
metastatic	O
hepatocellular	B-Cell
carcinoma	I-Cell
cell	I-Cell
lines	I-Cell
displayed	O
obvious	O
differences	O
compared	O
with	O
non	O
-	O
metastatic	O
liver	B-Cell
cancer	I-Cell
cell	I-Cell
lines	I-Cell
.	O

The	O
results	O
imply	O
that	O
various	O
different	O
proteins	O
may	O
lead	O
to	O
HCC	B-Cancer
metastasis	O
together	O
.	O

Clinical	O
implication	O
of	O
expression	O
of	O
cyclooxygenase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
peroxisome	B-Gene_or_gene_product
proliferator	I-Gene_or_gene_product
activated	I-Gene_or_gene_product
-	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
gamma	I-Gene_or_gene_product
in	O
epithelial	B-Cancer
ovarian	I-Cancer
tumours	I-Cancer
.	O

Expression	O
of	O
cyclooxygenase	B-Gene_or_gene_product
(	I-Gene_or_gene_product
COX	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
plays	O
a	O
key	O
role	O
in	O
tumorigenesis	O
and	O
development	O
and	O
peroxisome	B-Gene_or_gene_product
proliferator	I-Gene_or_gene_product
-	I-Gene_or_gene_product
activated	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
gamma	I-Gene_or_gene_product
(	O
PPARgamma	B-Gene_or_gene_product
)	O
has	O
been	O
implicated	O
in	O
the	O
control	O
of	O
COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
expression	O
in	O
some	O
tissues	B-Tissue
.	O

The	O
aim	O
of	O
this	O
study	O
is	O
to	O
investigate	O
(	O
1	O
)	O
whether	O
expression	O
of	O
COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
PPARgamma	B-Gene_or_gene_product
is	O
associated	O
with	O
ovarian	B-Organ
carcinogenesis	O
and	O
progression	O
of	O
ovarian	B-Cancer
tumours	I-Cancer
and	O
(	O
2	O
)	O
whether	O
COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
expression	O
is	O
controlled	O
through	O
ligand	O
-	O
mediated	O
activation	O
of	O
PPARgamma	B-Gene_or_gene_product
in	O
ovarian	B-Cell
carcinoma	I-Cell
cells	I-Cell
.	O

For	O
this	O
purpose	O
,	O
the	O
presence	O
of	O
COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
PPARgamma	B-Gene_or_gene_product
was	O
immunohistochemically	O
examined	O
in	O
71	O
epithelial	B-Cancer
ovarian	I-Cancer
carcinomas	I-Cancer
,	O
18	O
borderline	O
tumours	B-Cancer
and	O
23	O
benign	O
tumours	B-Cancer
and	O
the	O
levels	O
of	O
COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
PPARgamma	B-Gene_or_gene_product
proteins	O
were	O
determined	O
by	O
enzyme	O
immunoassay	O
in	O
four	O
benign	O
tumours	B-Cancer
,	O
three	O
borderline	O
tumours	B-Cancer
and	O
12	O
carcinomas	B-Cancer
.	O

The	O
frequency	O
of	O
COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
PPARgamma	B-Gene_or_gene_product
detection	O
was	O
significantly	O
increased	O
and	O
decreased	O
as	O
lesions	B-Pathological_formation
progressed	O
to	O
carcinoma	B-Cancer
,	O
respectively	O
.	O

The	O
COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
protein	O
was	O
not	O
detected	O
in	O
the	O
three	O
borderline	O
tumours	B-Cancer
,	O
whereas	O
PPARgamma	B-Gene_or_gene_product
protein	O
was	O
detected	O
in	O
all	O
of	O
them	O
.	O

COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
protein	O
was	O
detected	O
in	O
eight	O
of	O
the	O
12	O
carcinomas	B-Cancer
,	O
whereas	O
PPARgamma	B-Gene_or_gene_product
protein	O
was	O
detected	O
in	O
only	O
two	O
cases	O
.	O

In	O
addition	O
,	O
PPARgamma	B-Gene_or_gene_product
protein	O
was	O
not	O
detected	O
in	O
all	O
of	O
the	O
eight	O
carcinomas	B-Cancer
in	O
which	O
COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
protein	O
was	O
detected	O
,	O
suggesting	O
that	O
expression	O
of	O
PPARgamma	B-Gene_or_gene_product
and	O
COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
was	O
in	O
a	O
reciprocal	O
relationship	O
.	O

Furthermore	O
,	O
in	O
cultured	O
ovarian	B-Cell
carcinoma	I-Cell
cells	I-Cell
,	O
Western	O
blot	O
revealed	O
that	O
PPARgamma	B-Gene_or_gene_product
and	O
COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
expression	O
was	O
regulated	O
conversely	O
as	O
a	O
result	O
of	O
stimulation	O
by	O
15	B-Simple_chemical
-	I-Simple_chemical
deoxy	I-Simple_chemical
-	I-Simple_chemical
Delta	I-Simple_chemical
(	I-Simple_chemical
12	I-Simple_chemical
,	I-Simple_chemical
14	I-Simple_chemical
)	I-Simple_chemical
PGJ	I-Simple_chemical
(	I-Simple_chemical
2	I-Simple_chemical
)	I-Simple_chemical
(	O
15	B-Simple_chemical
-	I-Simple_chemical
PGJ	I-Simple_chemical
(	I-Simple_chemical
2	I-Simple_chemical
)	I-Simple_chemical
)	O
,	O
a	O
PPARgamma	B-Gene_or_gene_product
activator	O
.	O

In	O
addition	O
,	O
15d	B-Simple_chemical
-	I-Simple_chemical
PGJ	I-Simple_chemical
(	I-Simple_chemical
2	I-Simple_chemical
)	I-Simple_chemical
suppressed	O
tumour	B-Gene_or_gene_product
necrosis	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
-	O
induced	O
-	O
COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
expression	O
,	O
confirming	O
the	O
reciprocal	O
correlation	O
between	O
COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
PPARgamma	B-Gene_or_gene_product
.	O

From	O
these	O
results	O
,	O
it	O
was	O
suggested	O
that	O
PPARgamma	B-Gene_or_gene_product
activation	O
might	O
suppress	O
COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
expression	O
via	O
the	O
nuclear	B-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
pathway	O
in	O
the	O
ovarian	B-Cell
carcinoma	I-Cell
cells	I-Cell
and	O
that	O
low	O
expression	O
of	O
PPARgamma	B-Gene_or_gene_product
and	O
high	O
expression	O
of	O
COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
might	O
be	O
involved	O
in	O
carcinogenesis	O
and	O
progression	O
of	O
ovarian	B-Cancer
tumours	I-Cancer
.	O

Role	O
of	O
osteopontin	B-Gene_or_gene_product
in	O
adhesion	O
,	O
migration	O
,	O
cell	B-Cell
survival	O
and	O
bone	B-Organ
remodeling	O
.	O

Osteopontin	B-Gene_or_gene_product
(	O
OPN	B-Gene_or_gene_product
)	O
is	O
a	O
secreted	O
adhesive	O
glycophosphoprotein	O
expressed	O
by	O
several	O
cell	B-Cell
types	O
.	O

It	O
is	O
normally	O
produced	O
in	O
bone	B-Tissue
,	O
teeth	B-Organ
,	O
kidney	B-Organ
and	O
epithelial	B-Tissue
lining	I-Tissue
tissues	I-Tissue
and	O
is	O
found	O
in	O
plasma	B-Organism_substance
and	O
breast	B-Organism_substance
milk	I-Organism_substance
.	O

It	O
is	O
involved	O
in	O
a	O
number	O
of	O
physiologic	O
and	O
pathologic	O
events	O
including	O
angiogenesis	O
,	O
apoptosis	O
,	O
inflammation	O
,	O
wound	B-Pathological_formation
healing	O
and	O
tumor	B-Cancer
metastasis	O
.	O

In	O
this	O
review	O
focus	O
will	O
be	O
on	O
OPN	B-Gene_or_gene_product
in	O
bone	B-Tissue
and	O
its	O
role	O
in	O
adhesion	O
,	O
migration	O
and	O
cell	B-Cell
survival	O
.	O

These	O
aspects	O
of	O
OPN	B-Gene_or_gene_product
biology	O
are	O
important	O
in	O
tumorigenesis	O
.	O

Oxidation	O
in	O
rheumatoid	O
arthritis	O
.	O

Oxygen	B-Simple_chemical
metabolism	O
has	O
an	O
important	O
role	O
in	O
the	O
pathogenesis	O
of	O
rheumatoid	O
arthritis	O
.	O

Reactive	B-Simple_chemical
oxygen	I-Simple_chemical
species	I-Simple_chemical
(	O
ROS	B-Simple_chemical
)	O
produced	O
in	O
the	O
course	O
of	O
cellular	B-Cell
oxidative	O
phosphorylation	O
,	O
and	O
by	O
activated	O
phagocytic	B-Cell
cells	I-Cell
during	O
oxidative	O
bursts	O
,	O
exceed	O
the	O
physiological	O
buffering	O
capacity	O
and	O
result	O
in	O
oxidative	O
stress	O
.	O

The	O
excessive	O
production	O
of	O
ROS	B-Simple_chemical
can	O
damage	O
protein	O
,	O
lipids	B-Simple_chemical
,	O
nucleic	B-Simple_chemical
acids	I-Simple_chemical
,	O
and	O
matrix	B-Cellular_component
components	I-Cellular_component
.	O

They	O
also	O
serve	O
as	O
important	O
intracellular	B-Immaterial_anatomical_entity
signaling	O
molecules	O
that	O
amplify	O
the	O
synovial	B-Multi-tissue_structure
inflammatory	O
-	O
proliferative	O
response	O
.	O

Repetitive	O
cycles	O
of	O
hypoxia	O
and	O
reoxygenation	O
associated	O
with	O
changes	O
in	O
synovial	B-Multi-tissue_structure
perfusion	O
are	O
postulated	O
to	O
activate	O
hypoxia	B-Gene_or_gene_product
-	I-Gene_or_gene_product
inducible	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
and	O
nuclear	B-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
,	O
two	O
key	O
transcription	O
factors	O
that	O
are	O
regulated	O
by	O
changes	O
in	O
cellular	B-Cell
oxygenation	O
and	O
cytokine	O
stimulation	O
,	O
and	O
that	O
in	O
turn	O
orchestrate	O
the	O
expression	O
of	O
a	O
spectrum	O
of	O
genes	O
critical	O
to	O
the	O
persistence	O
of	O
synovitis	O
.	O

An	O
understanding	O
of	O
the	O
complex	O
interactions	O
involved	O
in	O
these	O
pathways	O
might	O
allow	O
the	O
development	O
of	O
novel	O
therapeutic	O
strategies	O
for	O
rheumatoid	O
arthritis	O
.	O

Glucocorticoid	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
-	O
induced	O
MAPK	B-Gene_or_gene_product
phosphatase	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
MPK	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
expression	O
inhibits	O
paclitaxel	B-Simple_chemical
-	O
associated	O
MAPK	B-Gene_or_gene_product
activation	O
and	O
contributes	O
to	O
breast	B-Cell
cancer	I-Cell
cell	I-Cell
survival	O
.	O

Glucocorticoid	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
(	O
GR	B-Gene_or_gene_product
)	O
activation	O
has	O
recently	O
been	O
shown	O
to	O
inhibit	O
apoptosis	O
in	O
breast	B-Cell
epithelial	I-Cell
cells	I-Cell
.	O

We	O
have	O
previously	O
described	O
a	O
group	O
of	O
genes	O
that	O
is	O
rapidly	O
up	O
-	O
regulated	O
in	O
these	O
cells	B-Cell
following	O
dexamethasone	B-Simple_chemical
(	O
Dex	B-Simple_chemical
)	O
treatment	O
.	O

In	O
an	O
effort	O
to	O
dissect	O
the	O
mechanisms	O
of	O
GR	B-Gene_or_gene_product
-	O
mediated	O
breast	B-Cell
epithelial	I-Cell
cell	I-Cell
survival	O
,	O
we	O
now	O
examine	O
the	O
molecular	O
events	O
downstream	O
of	O
GR	B-Gene_or_gene_product
activation	O
.	O

Here	O
we	O
show	O
that	O
GR	B-Gene_or_gene_product
activation	O
leads	O
to	O
both	O
the	O
rapid	O
induction	O
of	O
MAPK	B-Gene_or_gene_product
phosphatase	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
MKP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
mRNA	O
and	O
its	O
sustained	O
expression	O
.	O

Induction	O
of	O
the	O
MKP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
protein	O
in	O
the	O
MCF10A	B-Cell
-	I-Cell
Myc	I-Cell
and	O
MDA	B-Cell
-	I-Cell
MB	I-Cell
-	I-Cell
231	I-Cell
breast	I-Cell
epithelial	I-Cell
cell	I-Cell
lines	I-Cell
was	O
also	O
seen	O
.	O

Paclitaxel	B-Simple_chemical
treatment	O
resulted	O
in	O
MAPK	B-Gene_or_gene_product
activation	O
and	O
apoptosis	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
,	O
and	O
both	O
processes	O
were	O
inhibited	O
by	O
Dex	B-Simple_chemical
pretreatment	O
.	O

Furthermore	O
,	O
induction	O
of	O
MKP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
correlated	O
with	O
the	O
inhibition	O
of	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
ERK1	B-Gene_or_gene_product
/	I-Gene_or_gene_product
2	I-Gene_or_gene_product
)	O
and	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
(	O
JNK	B-Gene_or_gene_product
)	O
activity	O
,	O
whereas	O
p38	B-Gene_or_gene_product
activity	O
was	O
minimally	O
affected	O
.	O

Blocking	O
Dex	B-Simple_chemical
-	O
induced	O
MKP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
induction	O
using	O
small	O
interfering	O
RNA	O
increased	O
ERK1	B-Gene_or_gene_product
/	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
JNK	B-Gene_or_gene_product
phosphorylation	O
and	O
decreased	O
cell	B-Cell
survival	O
.	O

ERK1	B-Gene_or_gene_product
/	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
JNK	B-Gene_or_gene_product
inactivation	O
was	O
associated	O
with	O
Ets	B-Gene_or_gene_product
-	I-Gene_or_gene_product
like	I-Gene_or_gene_product
transcription	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
ELK	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
dephosphorylation	O
.	O

To	O
explore	O
the	O
gene	O
expression	O
changes	O
that	O
occur	O
downstream	O
of	O
ELK	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
dephosphorylation	O
,	O
we	O
used	O
a	O
combination	O
of	O
temporal	O
gene	O
expression	O
data	O
and	O
promoter	O
element	O
analyses	O
.	O

This	O
approach	O
revealed	O
a	O
previously	O
unrecognized	O
transcriptional	O
target	O
of	O
ELK	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
the	O
human	B-Organism
tissue	B-Gene_or_gene_product
plasminogen	I-Gene_or_gene_product
activator	I-Gene_or_gene_product
(	O
tPA	B-Gene_or_gene_product
)	O
.	O

We	O
verified	O
the	O
predicted	O
ELK	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
-	O
>	O
tPA	B-Gene_or_gene_product
transcriptional	O
regulatory	O
relationship	O
using	O
a	O
luciferase	B-Gene_or_gene_product
reporter	O
assay	O
.	O

We	O
conclude	O
that	O
GR	B-Gene_or_gene_product
-	O
mediated	O
MAPK	B-Gene_or_gene_product
inactivation	O
contributes	O
to	O
cell	B-Cell
survival	O
and	O
that	O
the	O
potential	O
transcriptional	O
targets	O
of	O
this	O
inhibition	O
can	O
be	O
identified	O
from	O
large	O
scale	O
gene	O
array	O
analysis	O
.	O

Investigating	O
the	O
causes	O
of	O
low	O
birth	O
weight	O
in	O
contrasting	O
ovine	O
paradigms	O
.	O

Intrauterine	O
growth	O
restriction	O
(	O
IUGR	O
)	O
still	O
accounts	O
for	O
a	O
large	O
incidence	O
of	O
infant	B-Organism
mortality	O
and	O
morbidity	O
worldwide	O
.	O

Many	O
of	O
the	O
circulatory	O
and	O
transport	O
properties	O
of	O
the	O
sheep	B-Organism
placenta	B-Organ
are	O
similar	O
to	O
those	O
of	O
the	O
human	B-Organism
placenta	B-Organ
and	O
as	O
such	O
,	O
the	O
pregnant	O
sheep	B-Organism
offers	O
an	O
excellent	O
model	O
in	O
which	O
to	O
study	O
the	O
development	O
of	O
IUGR	O
.	O

Two	O
natural	O
models	O
of	O
ovine	O
IUGR	O
are	O
those	O
of	O
hyperthermic	O
exposure	O
during	O
pregnancy	O
,	O
and	O
adolescent	O
overfeeding	O
,	O
also	O
during	O
pregnancy	O
.	O

Both	O
models	O
yield	O
significantly	O
reduced	O
placental	B-Organ
weights	O
and	O
an	O
asymmetrically	O
growth	O
-	O
restricted	O
fetus	B-Developing_anatomical_structure
,	O
and	O
display	O
altered	O
maternal	O
hormone	O
concentrations	O
,	O
indicative	O
of	O
an	O
impaired	O
trophoblast	B-Cell
capacity	O
.	O

Additionally	O
,	O
impaired	O
placental	B-Organ
angiogenesis	O
and	O
uteroplacental	B-Organism_substance
blood	I-Organism_substance
flow	O
appears	O
to	O
be	O
an	O
early	O
defect	O
in	O
both	O
the	O
hyperthermic	O
and	O
adolescent	O
paradigms	O
.	O

The	O
effects	O
of	O
these	O
alterations	O
in	O
placental	B-Organ
functional	O
development	O
appear	O
to	O
be	O
irreversible	O
.	O

IUGR	O
fetuses	B-Developing_anatomical_structure
are	O
both	O
hypoxic	O
and	O
hypoglycaemic	O
,	O
and	O
have	O
reduced	O
insulin	B-Gene_or_gene_product
and	O
insulin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
like	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
IGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
,	O
and	O
elevated	O
concentrations	O
of	O
lactate	B-Simple_chemical
.	O

However	O
,	O
fetal	B-Developing_anatomical_structure
utilization	O
of	O
oxygen	B-Simple_chemical
and	O
glucose	B-Simple_chemical
,	O
on	O
a	O
weight	O
basis	O
,	O
remain	O
constant	O
compared	O
with	O
control	O
pregnancies	O
.	O

Maintained	O
utilization	O
of	O
these	O
substrates	O
,	O
in	O
a	O
substrate	O
-	O
deficient	O
environment	O
,	O
suggests	O
increased	O
sensitivities	O
to	O
metabolic	O
signals	O
,	O
which	O
may	O
play	O
a	O
role	O
in	O
the	O
development	O
of	O
metabolic	O
diseases	O
in	O
later	O
adult	O
life	O
.	O

p53	B-Gene_or_gene_product
mutation	O
heterogeneity	O
in	O
cancer	B-Cancer
.	O

The	O
p53	B-Gene_or_gene_product
gene	O
is	O
inactivated	O
in	O
about	O
50	O
%	O
of	O
human	B-Organism
cancers	B-Cancer
and	O
the	O
p53	B-Gene_or_gene_product
protein	O
is	O
an	O
essential	O
component	O
of	O
the	O
cell	B-Cell
response	O
induced	O
by	O
genotoxic	O
stresses	O
such	O
as	O
those	O
generated	O
by	O
radiotherapy	O
or	O
chemotherapy	O
.	O

It	O
is	O
therefore	O
highly	O
likely	O
that	O
these	O
alterations	O
are	O
an	O
important	O
component	O
in	O
tumor	B-Cancer
resistance	O
to	O
therapy	O
.	O

The	O
particular	O
characteristics	O
of	O
these	O
alterations	O
,	O
80	O
%	O
of	O
which	O
are	O
missense	O
mutations	O
leading	O
to	O
functionally	O
heterogeneous	O
proteins	O
,	O
make	O
p53	B-Gene_or_gene_product
a	O
unique	O
gene	O
in	O
the	O
class	O
of	O
tumor	B-Cancer
suppressor	O
genes	O
.	O

A	O
considerable	O
number	O
of	O
mutant	O
p53	B-Gene_or_gene_product
proteins	O
probably	O
have	O
an	O
oncogenic	O
activity	O
per	O
se	O
and	O
therefore	O
actively	O
participate	O
in	O
cell	B-Cell
transformation	O
.	O

The	O
fact	O
that	O
the	O
apoptotic	O
and	O
antiproliferative	O
functions	O
of	O
p53	B-Gene_or_gene_product
can	O
be	O
dissociated	O
in	O
certain	O
mutants	O
also	O
suggests	O
another	O
level	O
of	O
complexity	O
in	O
the	O
relationships	O
between	O
p53	B-Gene_or_gene_product
inactivation	O
and	O
neoplasia	B-Cancer
.	O

Randomized	O
,	O
open	O
label	O
,	O
prospective	O
study	O
on	O
the	O
effect	O
of	O
zoledronic	B-Simple_chemical
acid	I-Simple_chemical
on	O
the	O
prevention	O
of	O
bone	B-Cancer
metastases	I-Cancer
in	O
patients	B-Organism
with	O
recurrent	O
solid	B-Cancer
tumors	I-Cancer
that	O
did	O
not	O
present	O
with	O
bone	B-Cancer
metastases	I-Cancer
at	O
baseline	O
.	O

OBJECTIVES	O
:	O
Bisphosphonates	B-Simple_chemical
have	O
been	O
used	O
successfully	O
in	O
the	O
treatment	O
of	O
hypercalcemia	O
and	O
to	O
reduce	O
skeletal	B-Anatomical_system
-	O
related	O
complications	O
of	O
bone	B-Cancer
metastases	I-Cancer
.	O

Recent	O
in	O
vitro	O
and	O
in	O
vivo	O
evidence	O
suggest	O
that	O
they	O
may	O
also	O
have	O
direct	O
antitumor	B-Cancer
effects	O
via	O
induction	O
of	O
apoptosis	O
,	O
inhibition	O
of	O
the	O
invasive	O
potential	O
of	O
tumor	B-Cell
cell	I-Cell
lines	I-Cell
in	O
vitro	O
,	O
inhibition	O
of	O
angiogenesis	O
,	O
and	O
reduction	O
in	O
tumor	B-Cancer
growth	O
indirectly	O
via	O
effects	O
on	O
accessory	B-Cell
cells	I-Cell
.	O

This	O
is	O
a	O
randomized	O
,	O
open	O
label	O
,	O
prospective	O
study	O
that	O
examined	O
the	O
effect	O
of	O
preventive	O
zoledronic	B-Simple_chemical
acid	I-Simple_chemical
treatment	O
on	O
the	O
development	O
of	O
bone	B-Cancer
metastases	I-Cancer
in	O
patients	B-Organism
with	O
recurrent	O
solid	B-Cancer
tumors	I-Cancer
,	O
without	O
bone	B-Cancer
metastases	I-Cancer
at	O
the	O
time	O
of	O
randomization	O
.	O

METHODS	O
:	O
Forty	O
patients	B-Organism
with	O
recurrent	O
or	O
metastatic	O
advanced	O
cancer	B-Cancer
,	O
without	O
bone	B-Cancer
metastases	I-Cancer
,	O
were	O
randomized	O
into	O
the	O
trial	O
to	O
either	O
receive	O
zoledronic	B-Simple_chemical
acid	I-Simple_chemical
or	O
no	O
treatment	O
.	O

Patients	B-Organism
were	O
followed	O
up	O
until	O
bone	B-Cancer
metastases	I-Cancer
were	O
established	O
.	O

RESULTS	O
:	O
The	O
percentage	O
of	O
patients	B-Organism
being	O
bone	B-Cancer
metastases	I-Cancer
free	O
at	O
12	O
mo	O
was	O
60	O
%	O
in	O
the	O
zoledronic	B-Simple_chemical
acid	I-Simple_chemical
and	O
10	O
%	O
in	O
the	O
control	O
group	O
(	O
p	O
less	O
than	O
0	O
.	O
0005	O
)	O
,	O
while	O
the	O
percentages	O
at	O
18	O
mo	O
were	O
20	O
%	O
and	O
5	O
%	O
respectively	O
(	O
p	O
=	O
0	O
.	O
0002	O
)	O
.	O

CONCLUSIONS	O
:	O
The	O
results	O
have	O
shown	O
that	O
bisphosphonates	B-Simple_chemical
as	O
adjuvant	O
treatment	O
might	O
be	O
useful	O
for	O
the	O
prevention	O
of	O
bone	B-Cancer
metastases	I-Cancer
;	O
however	O
,	O
there	O
is	O
need	O
for	O
blinded	O
randomized	O
data	O
before	O
such	O
an	O
approach	O
would	O
be	O
confirmed	O
.	O

In	O
the	O
meantime	O
preventive	O
use	O
of	O
bisphosphonates	B-Simple_chemical
in	O
patients	B-Organism
without	O
any	O
bone	B-Cancer
metastases	I-Cancer
should	O
not	O
be	O
used	O
outside	O
the	O
scope	O
of	O
a	O
clinical	O
trial	O
.	O

Impaired	O
coronary	B-Multi-tissue_structure
collateral	I-Multi-tissue_structure
vessel	I-Multi-tissue_structure
development	O
in	O
patients	B-Organism
with	O
proliferative	O
diabetic	O
retinopathy	O
.	O

BACKGROUND	O
:	O
Diabetic	O
patients	B-Organism
have	O
been	O
reported	O
to	O
have	O
impaired	O
coronary	B-Multi-tissue_structure
collateral	I-Multi-tissue_structure
vessel	I-Multi-tissue_structure
growth	O
,	O
although	O
they	O
have	O
excessive	O
neovascularization	O
in	O
the	O
retina	B-Multi-tissue_structure
.	O

HYPOTHESIS	O
:	O
This	O
study	O
was	O
designed	O
to	O
compare	O
coronary	B-Multi-tissue_structure
collateral	I-Multi-tissue_structure
circulation	O
(	O
CCC	O
)	O
in	O
patients	B-Organism
with	O
proliferative	O
diabetic	O
retinopathy	O
(	O
PDR	O
)	O
with	O
that	O
in	O
patients	B-Organism
without	O
DR	O
.	O

METHODS	O
:	O
Ninety	O
diabetic	O
patients	B-Organism
with	O
chronic	O
total	O
occlusion	O
in	O
at	O
least	O
one	O
major	O
epicardial	B-Multi-tissue_structure
coronary	I-Multi-tissue_structure
artery	I-Multi-tissue_structure
were	O
enrolled	O
in	O
the	O
study	O
.	O

Groups	O
1	O
and	O
2	O
consisted	O
of	O
48	O
patients	B-Organism
without	O
DR	O
and	O
42	O
patients	B-Organism
with	O
PDR	O
,	O
respectively	O
.	O

Coronary	B-Multi-tissue_structure
collateral	I-Multi-tissue_structure
circulation	O
(	O
CCC	O
)	O
was	O
analyzed	O
according	O
to	O
the	O
Rentrop	O
system	O
.	O

Each	O
group	O
was	O
also	O
divided	O
into	O
two	O
subgroups	O
according	O
to	O
poor	O
and	O
good	O
CCC	O
.	O

Serum	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
(	O
VEGF	B-Gene_or_gene_product
)	O
levels	O
were	O
measured	O
using	O
the	O
enzyme	O
-	O
linked	O
immunosorbent	O
assay	O
(	O
ELISA	O
)	O
kit	O
.	O

RESULTS	O
:	O
The	O
mean	O
Rentrop	O
collateral	O
score	O
was	O
higher	O
in	O
Group	O
1	O
than	O
in	O
Group	O
2	O
(	O
2	O
.	O
39	O
+	O
/	O
-	O
1	O
.	O
07	O
vs	O
.	O
1	O
.	O
76	O
+	O
/	O
-	O
0	O
.	O
76	O
,	O
respectively	O
,	O
p	O
less	O
than	O
0	O
.	O
001	O
)	O
.	O

When	O
the	O
two	O
groups	O
were	O
compared	O
with	O
respect	O
to	O
poor	O
and	O
good	O
CCC	O
,	O
poor	O
CCC	O
was	O
higher	O
in	O
patients	B-Organism
with	O
PDR	O
(	O
64	O
vs	O
.	O
36	O
%	O
,	O
respectively	O
,	O
p	O
=	O
0	O
.	O
01	O
)	O
.	O

Serum	B-Organism_substance
VEGF	B-Gene_or_gene_product
levels	O
were	O
higher	O
in	O
patients	B-Organism
with	O
PDR	O
than	O
in	O
those	O
without	O
DR	O
(	O
219	O
+	O
/	O
-	O
99	O
vs	O
.	O
139	O
+	O
/	O
-	O
98	O
pg	O
/	O
ml	O
,	O
p	O
less	O
than	O
0	O
.	O
001	O
)	O
;	O
however	O
,	O
patients	B-Organism
with	O
poor	O
and	O
good	O
CCC	O
had	O
similar	O
VEGF	B-Gene_or_gene_product
levels	O
.	O

CONCLUSIONS	O
:	O
We	O
have	O
shown	O
that	O
patients	B-Organism
with	O
PDR	O
have	O
a	O
lower	O
coronary	B-Multi-tissue_structure
collateral	I-Multi-tissue_structure
score	O
than	O
patients	B-Organism
without	O
DR	O
.	O

Also	O
,	O
serum	B-Organism_substance
VEGF	B-Gene_or_gene_product
was	O
significantly	O
higher	O
in	O
patients	B-Organism
with	O
PDR	O
than	O
in	O
those	O
without	O
DR	O
.	O

These	O
findings	O
have	O
suggested	O
that	O
diabetes	O
mellitus	O
may	O
have	O
a	O
different	O
action	O
on	O
retinal	B-Multi-tissue_structure
and	O
coronary	B-Multi-tissue_structure
circulation	O
.	O

VEGFR1	B-Gene_or_gene_product
-	O
positive	O
haematopoietic	O
bone	O
marrow	O
progenitors	O
initiate	O
the	O
pre	O
-	O
metastatic	O
niche	O
.	O

The	O
cellular	B-Cell
and	O
molecular	O
mechanisms	O
by	O
which	O
a	O
tumour	B-Cell
cell	I-Cell
undergoes	O
metastasis	O
to	O
a	O
predetermined	O
location	O
are	O
largely	O
unknown	O
.	O

Here	O
we	O
demonstrate	O
that	O
bone	B-Cell
marrow	I-Cell
-	I-Cell
derived	I-Cell
haematopoietic	I-Cell
progenitor	I-Cell
cells	I-Cell
that	O
express	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
1	I-Gene_or_gene_product
(	O
VEGFR1	B-Gene_or_gene_product
;	O
also	O
known	O
as	O
Flt1	B-Gene_or_gene_product
)	O
home	O
to	O
tumour	B-Cancer
-	O
specific	O
pre	O
-	O
metastatic	O
sites	B-Multi-tissue_structure
and	O
form	O
cellular	B-Cell
clusters	I-Cell
before	O
the	O
arrival	O
of	O
tumour	B-Cell
cells	I-Cell
.	O

Preventing	O
VEGFR1	B-Gene_or_gene_product
function	O
using	O
antibodies	O
or	O
by	O
the	O
removal	O
of	O
VEGFR1	B-Gene_or_gene_product
(	O
+	O
)	O
cells	O
from	O
the	O
bone	B-Multi-tissue_structure
marrow	I-Multi-tissue_structure
of	O
wild	O
-	O
type	O
mice	B-Organism
abrogates	O
the	O
formation	O
of	O
these	O
pre	O
-	O
metastatic	O
clusters	B-Cell
and	O
prevents	O
tumour	B-Cancer
metastasis	O
,	O
whereas	O
reconstitution	O
with	O
selected	O
Id3	B-Gene_or_gene_product
(	O
inhibitor	B-Gene_or_gene_product
of	I-Gene_or_gene_product
differentiation	I-Gene_or_gene_product
3	I-Gene_or_gene_product
)	O
-	O
competent	O
VEGFR1	B-Gene_or_gene_product
+	O
cells	O
establishes	O
cluster	B-Cell
formation	O
and	O
tumour	B-Cancer
metastasis	O
in	O
Id3	B-Gene_or_gene_product
knockout	O
mice	O
.	O

We	O
also	O
show	O
that	O
VEGFR1	B-Gene_or_gene_product
+	O
cells	O
express	O
VLA	B-Gene_or_gene_product
-	I-Gene_or_gene_product
4	I-Gene_or_gene_product
(	O
also	O
known	O
as	O
integrin	B-Gene_or_gene_product
alpha4beta1	I-Gene_or_gene_product
)	O
,	O
and	O
that	O
tumour	B-Cancer
-	O
specific	O
growth	O
factors	O
upregulate	O
fibronectin	B-Gene_or_gene_product
-	O
-	O
a	O
VLA	B-Gene_or_gene_product
-	I-Gene_or_gene_product
4	I-Gene_or_gene_product
ligand	O
-	O
-	O
in	O
resident	O
fibroblasts	B-Cell
,	O
providing	O
a	O
permissive	O
niche	O
for	O
incoming	O
tumour	B-Cell
cells	I-Cell
.	O

Conditioned	O
media	O
obtained	O
from	O
distinct	O
tumour	B-Cancer
types	O
with	O
unique	O
patterns	O
of	O
metastatic	O
spread	O
redirected	O
fibronectin	B-Gene_or_gene_product
expression	O
and	O
cluster	B-Cell
formation	O
,	O
thereby	O
transforming	O
the	O
metastatic	O
profile	O
.	O

These	O
findings	O
demonstrate	O
a	O
requirement	O
for	O
VEGFR1	B-Gene_or_gene_product
+	O
haematopoietic	O
progenitors	O
in	O
the	O
regulation	O
of	O
metastasis	O
,	O
and	O
suggest	O
that	O
expression	O
patterns	O
of	O
fibronectin	B-Gene_or_gene_product
and	O
VEGFR1	B-Gene_or_gene_product
+	O
VLA	B-Gene_or_gene_product
-	I-Gene_or_gene_product
4	I-Gene_or_gene_product
+	O
clusters	O
dictate	O
organ	B-Organ
-	O
specific	O
tumour	B-Cancer
spread	O
.	O

The	O
Kaposi	B-Organism
'	I-Organism
s	I-Organism
sarcoma	I-Organism
-	I-Organism
associated	I-Organism
herpesvirus	I-Organism
G	B-Gene_or_gene_product
protein	I-Gene_or_gene_product
-	I-Gene_or_gene_product
coupled	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
as	O
a	O
therapeutic	O
target	O
for	O
the	O
treatment	O
of	O
Kaposi	B-Cancer
'	I-Cancer
s	I-Cancer
sarcoma	I-Cancer
.	O

The	O
Kaposi	B-Organism
'	I-Organism
s	I-Organism
sarcoma	I-Organism
-	I-Organism
associated	I-Organism
herpesvirus	I-Organism
(	O
KSHV	B-Organism
)	O
encodes	O
a	O
G	B-Gene_or_gene_product
protein	I-Gene_or_gene_product
-	I-Gene_or_gene_product
coupled	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
(	O
vGPCR	B-Gene_or_gene_product
)	O
that	O
has	O
been	O
implicated	O
in	O
the	O
initiation	O
of	O
Kaposi	B-Cancer
'	I-Cancer
s	I-Cancer
sarcoma	I-Cancer
,	O
identifying	O
vGPCR	B-Gene_or_gene_product
as	O
an	O
attractive	O
target	O
for	O
preventing	O
Kaposi	B-Cancer
'	I-Cancer
s	I-Cancer
sarcoma	I-Cancer
.	O

However	O
,	O
as	O
only	O
a	O
fraction	O
of	O
cells	B-Cell
in	O
advanced	B-Cancer
Kaposi	I-Cancer
'	I-Cancer
s	I-Cancer
sarcoma	I-Cancer
lesions	I-Cancer
express	O
vGPCR	B-Gene_or_gene_product
,	O
it	O
is	O
unclear	O
whether	O
this	O
unique	O
viral	O
oncogene	O
contributes	O
to	O
Kaposi	B-Cancer
'	I-Cancer
s	I-Cancer
sarcoma	I-Cancer
progression	O
.	O

We	O
therefore	O
set	O
out	O
to	O
determine	O
whether	O
the	O
few	O
cells	B-Cell
that	O
express	O
vGPCR	B-Gene_or_gene_product
in	O
established	O
tumors	B-Cancer
represent	O
an	O
appropriate	O
therapeutic	O
target	O
for	O
the	O
treatment	O
of	O
patients	B-Organism
with	O
preexisting	O
Kaposi	B-Cancer
'	I-Cancer
s	I-Cancer
sarcoma	I-Cancer
.	O

To	O
this	O
end	O
,	O
we	O
generated	O
endothelial	B-Cell
cell	I-Cell
lines	I-Cell
stably	O
expressing	O
vGPCR	B-Gene_or_gene_product
or	O
key	O
KSHV	B-Organism
latently	O
expressed	O
proteins	O
(	O
vCyclin	B-Gene_or_gene_product
,	O
vFlip	B-Gene_or_gene_product
,	O
and	O
LANA1	B-Gene_or_gene_product
)	O
.	O

The	O
endothelial	B-Cell
cell	I-Cell
line	I-Cell
expressing	O
vGPCR	B-Gene_or_gene_product
was	O
rendered	O
sensitive	O
to	O
treatment	O
with	O
the	O
nucleoside	B-Simple_chemical
analogue	O
ganciclovir	B-Simple_chemical
by	O
using	O
a	O
bicistronic	O
construct	O
coexpressing	O
the	O
herpes	B-Organism
simplex	I-Organism
virus	I-Organism
1	I-Organism
thymidine	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
.	O

S	O
.	O
c	O
.	O
injection	O
into	O
nude	B-Organism
mice	I-Organism
with	O
mixed	B-Cell
-	I-Cell
cell	I-Cell
populations	I-Cell
formed	O
tumors	B-Cancer
that	O
approximate	O
the	O
ratio	O
of	O
vGPCR	B-Gene_or_gene_product
-	O
expressing	O
and	O
KSHV	B-Organism
latent	O
gene	O
-	O
expressing	O
cells	B-Cell
.	O

These	O
mice	B-Organism
were	O
then	O
treated	O
with	O
ganciclovir	B-Simple_chemical
to	O
specifically	O
target	O
only	O
the	O
vGPCR	B-Gene_or_gene_product
-	O
expressing	O
cells	B-Cell
.	O

Surprisingly	O
,	O
despite	O
the	O
expression	O
of	O
KSHV	B-Organism
latent	O
genes	O
in	O
the	O
vast	O
majority	O
of	O
tumor	B-Cell
cells	I-Cell
,	O
specifically	O
targeting	O
only	O
the	O
few	O
vGPCR	B-Gene_or_gene_product
-	O
expressing	O
cells	B-Cell
in	O
established	O
tumors	B-Cancer
resulted	O
in	O
tumor	B-Cancer
regression	O
.	O

Moreover	O
,	O
we	O
observed	O
an	O
increase	O
in	O
apoptosis	O
of	O
latent	O
gene	O
-	O
expressing	O
cells	B-Cell
after	O
the	O
pharmacologic	O
deletion	O
of	O
the	O
vGPCR	B-Gene_or_gene_product
-	O
expressing	O
cells	B-Cell
.	O

These	O
findings	O
indicate	O
that	O
vGPCR	B-Gene_or_gene_product
may	O
play	O
a	O
key	O
role	O
in	O
Kaposi	B-Cancer
'	I-Cancer
s	I-Cancer
sarcoma	I-Cancer
progression	O
and	O
provide	O
experimental	O
justification	O
for	O
developing	O
molecular	O
-	O
based	O
therapies	O
specifically	O
targeting	O
vGPCR	B-Gene_or_gene_product
and	O
its	O
effectors	O
for	O
the	O
treatment	O
of	O
Kaposi	B-Cancer
'	I-Cancer
s	I-Cancer
sarcoma	I-Cancer
patients	B-Organism
.	O

Mechanisms	O
of	O
pericyte	B-Cell
recruitment	O
in	O
tumour	B-Cancer
angiogenesis	O
:	O
a	O
new	O
role	O
for	O
metalloproteinases	B-Gene_or_gene_product
.	O

Pericytes	B-Cell
occur	O
in	O
tumour	B-Multi-tissue_structure
blood	I-Multi-tissue_structure
vessels	I-Multi-tissue_structure
and	O
are	O
critical	O
for	O
the	O
development	O
of	O
a	O
functional	O
vascular	B-Anatomical_system
network	I-Anatomical_system
.	O

Targeting	O
tumour	B-Cell
pericytes	I-Cell
is	O
a	O
promising	O
anti	O
-	O
angiogenic	O
therapy	O
but	O
requires	O
identifying	O
the	O
mechanisms	O
of	O
their	O
recruitment	O
in	O
tumour	B-Cancer
and	O
addressing	O
whether	O
these	O
mechanisms	O
can	O
be	O
selectively	O
harnessed	O
.	O

Among	O
the	O
pathways	O
involved	O
in	O
pericyte	B-Cell
recruitment	O
during	O
embryonic	B-Developing_anatomical_structure
development	O
,	O
the	O
contribution	O
of	O
platelet	B-Gene_or_gene_product
-	I-Gene_or_gene_product
derived	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
B	I-Gene_or_gene_product
and	O
sphingosine	B-Gene_or_gene_product
1	I-Gene_or_gene_product
-	I-Gene_or_gene_product
phosphate	I-Gene_or_gene_product
is	O
confirmed	O
in	O
tumour	B-Cancer
angiogenesis	O
.	O

The	O
effect	O
of	O
angiopoietin	B-Gene_or_gene_product
1	I-Gene_or_gene_product
depends	O
on	O
the	O
tumour	B-Cancer
model	O
.	O

Transforming	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
beta1	I-Gene_or_gene_product
enhances	O
tumour	B-Cancer
vascularization	O
and	O
microvessel	B-Tissue
maturation	O
.	O

Recent	O
reports	O
suggest	O
a	O
participation	O
of	O
matrix	B-Gene_or_gene_product
metalloproteinases	I-Gene_or_gene_product
(	O
MMP	B-Gene_or_gene_product
)	O
in	O
tumour	B-Cell
pericyte	I-Cell
recruitment	O
that	O
is	O
consistent	O
with	O
the	O
effect	O
of	O
certain	O
MMPs	B-Gene_or_gene_product
in	O
the	O
development	O
of	O
microvasculature	B-Tissue
in	O
embryonic	B-Developing_anatomical_structure
development	O
and	O
in	O
in	O
vitro	O
models	O
of	O
vascular	B-Multi-tissue_structure
remodelling	O
.	O

Here	O
,	O
we	O
discuss	O
the	O
possibility	O
for	O
MMPs	B-Gene_or_gene_product
to	O
contribute	O
to	O
pericyte	B-Cell
recruitment	O
at	O
six	O
levels	O
:	O
(	O
1	O
)	O
direct	O
promotion	O
of	O
pericyte	B-Cell
invasion	O
by	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
degradation	O
;	O
(	O
2	O
)	O
stimulation	O
of	O
pericyte	B-Cell
proliferation	O
and	O
protection	O
against	O
apoptosis	O
by	O
modification	O
of	O
the	O
ECM	B-Cellular_component
;	O
(	O
3	O
)	O
activation	O
of	O
pericytes	B-Cell
through	O
the	O
release	O
of	O
growth	O
factor	O
bound	O
to	O
the	O
ECM	B-Cellular_component
;	O
(	O
4	O
)	O
transactivation	O
of	O
angiogenic	O
cell	B-Cellular_component
surface	I-Cellular_component
receptor	O
;	O
(	O
5	O
)	O
propagation	O
of	O
angiogenic	O
signalling	O
as	O
cofactor	O
;	O
and	O
(	O
6	O
)	O
recruitment	O
of	O
bone	B-Cell
marrow	I-Cell
-	I-Cell
derived	I-Cell
stem	I-Cell
cells	I-Cell
.	O

Sensitization	O
to	O
gimatecan	B-Simple_chemical
-	O
induced	O
apoptosis	O
by	O
tumor	B-Gene_or_gene_product
necrosis	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
related	I-Gene_or_gene_product
apoptosis	I-Gene_or_gene_product
inducing	I-Gene_or_gene_product
ligand	I-Gene_or_gene_product
in	O
prostate	B-Cell
carcinoma	I-Cell
cells	I-Cell
.	O

Since	O
the	O
intrinsic	O
resistance	O
of	O
prostate	B-Cancer
carcinoma	I-Cancer
likely	O
reflects	O
a	O
low	O
susceptibility	O
to	O
drug	O
-	O
induced	O
apoptosis	O
,	O
in	O
this	O
study	O
we	O
explored	O
the	O
possibility	O
of	O
sensitizing	O
prostate	B-Cell
carcinoma	I-Cell
cells	I-Cell
to	O
apoptosis	O
by	O
combination	O
of	O
TRAIL	B-Gene_or_gene_product
with	O
camptothecins	B-Simple_chemical
.	O

Indeed	O
,	O
these	O
agents	O
are	O
known	O
to	O
activate	O
different	O
pathways	O
of	O
apoptosis	O
.	O

Topotecan	B-Simple_chemical
-	O
and	O
gimatecan	B-Simple_chemical
induced	O
moderate	O
up	O
-	O
regulation	O
of	O
TRAIL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
R1	I-Gene_or_gene_product
and	O
-	B-Gene_or_gene_product
R2	I-Gene_or_gene_product
which	O
resulted	O
in	O
a	O
different	O
cell	B-Cell
response	O
to	O
the	O
combination	O
in	O
androgen	B-Simple_chemical
-	O
independent	O
cells	B-Cell
(	O
DU	B-Cell
-	I-Cell
145	I-Cell
and	O
PC	B-Cell
-	I-Cell
3	I-Cell
)	O
.	O

In	O
DU	B-Cell
-	I-Cell
145	I-Cell
cells	I-Cell
apoptosis	O
was	O
increased	O
by	O
lower	O
TRAIL	B-Gene_or_gene_product
concentrations	O
and	O
was	O
earlier	O
than	O
in	O
PC	B-Cell
-	I-Cell
3	I-Cell
cells	I-Cell
,	O
as	O
shown	O
using	O
Annexin	B-Gene_or_gene_product
V	I-Gene_or_gene_product
-	O
binding	O
assay	O
.	O

The	O
relative	O
resistance	O
of	O
PC	B-Cell
-	I-Cell
3	I-Cell
cells	I-Cell
to	O
drug	O
-	O
induced	O
apoptosis	O
was	O
associated	O
with	O
constitutive	O
Akt	B-Gene_or_gene_product
activation	O
,	O
higher	O
levels	O
of	O
cFLIP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
L	I-Gene_or_gene_product
and	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
and	O
lower	O
levels	O
of	O
Bax	B-Gene_or_gene_product
.	O

The	O
different	O
expression	O
/	O
activation	O
of	O
apoptosis	O
-	O
related	O
factors	O
appears	O
to	O
influence	O
the	O
sensitization	O
of	O
prostate	B-Cell
carcinoma	I-Cell
cells	I-Cell
by	O
TRAIL	B-Gene_or_gene_product
.	O

Potentiation	O
of	O
camptothecin	B-Simple_chemical
-	O
induced	O
apoptosis	O
by	O
TRAIL	B-Gene_or_gene_product
appears	O
dependent	O
on	O
cooperation	O
between	O
extrinsic	O
and	O
intrinsic	O
pathways	O
,	O
as	O
documented	O
by	O
loss	O
of	O
the	O
sensitization	O
to	O
apoptosis	O
following	O
reduction	O
of	O
caspase	B-Gene_or_gene_product
8	I-Gene_or_gene_product
after	O
small	O
interfering	O
RNA	O
transfection	O
.	O

The	O
efficacy	O
of	O
the	O
approach	O
may	O
be	O
critically	O
dependent	O
on	O
the	O
intrinsic	O
susceptibility	O
to	O
apoptosis	O
of	O
different	O
tumors	B-Cancer
.	O

These	O
observations	O
support	O
that	O
the	O
activation	O
of	O
multiple	O
signals	O
could	O
enhance	O
apoptotic	O
response	O
and	O
suggest	O
the	O
therapeutic	O
interest	O
of	O
the	O
TRAIL	B-Gene_or_gene_product
/	O
camptothecin	B-Simple_chemical
combination	O
.	O

c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Met	I-Gene_or_gene_product
expression	O
is	O
regulated	O
by	O
Mitf	B-Gene_or_gene_product
in	O
the	O
melanocyte	B-Cell
lineage	I-Cell
.	O

Hepatocyte	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
HGF	B-Gene_or_gene_product
)	O
/	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Met	I-Gene_or_gene_product
signaling	O
is	O
thought	O
to	O
be	O
a	O
key	O
pathway	O
in	O
both	O
melanocyte	B-Cell
development	O
and	O
melanoma	B-Cancer
metastasis	O
.	O

Here	O
,	O
HGF	B-Gene_or_gene_product
stimulation	O
of	O
melanocytes	B-Cell
was	O
seen	O
to	O
up	O
-	O
regulate	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Met	I-Gene_or_gene_product
expression	O
.	O

In	O
an	O
effort	O
to	O
decipher	O
the	O
mechanism	O
by	O
which	O
HGF	B-Gene_or_gene_product
up	O
-	O
regulates	O
its	O
receptor	O
,	O
we	O
found	O
that	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Met	I-Gene_or_gene_product
is	O
a	O
direct	O
transcriptional	O
target	O
of	O
Mitf	B-Gene_or_gene_product
.	O

This	O
was	O
confirmed	O
with	O
chromatin	B-Cellular_component
immunoprecipitation	O
experiments	O
of	O
the	O
human	B-Organism
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Met	I-Gene_or_gene_product
promoter	O
,	O
as	O
well	O
as	O
by	O
the	O
ability	O
of	O
adenovirally	B-Organism
expressed	O
Mitf	B-Gene_or_gene_product
to	O
modulate	O
endogenous	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Met	I-Gene_or_gene_product
protein	O
levels	O
in	O
melanocytes	B-Cell
.	O

Disruption	O
of	O
Mitf	B-Gene_or_gene_product
blocked	O
HGF	B-Gene_or_gene_product
-	O
dependent	O
increases	O
in	O
endogenous	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Met	I-Gene_or_gene_product
message	O
and	O
protein	O
levels	O
,	O
indicating	O
that	O
HGF	B-Gene_or_gene_product
regulates	O
its	O
own	O
receptor	O
levels	O
via	O
Mitf	B-Gene_or_gene_product
.	O

Finally	O
,	O
dominant	O
-	O
negative	O
inhibition	O
of	O
Mitf	B-Gene_or_gene_product
resulted	O
in	O
profound	O
resistance	O
of	O
melanocytes	B-Cell
and	O
melanoma	B-Cell
cells	I-Cell
to	O
HGF	B-Gene_or_gene_product
-	O
dependent	O
matrix	B-Cellular_component
invasion	O
,	O
suggesting	O
a	O
physiologic	O
role	O
for	O
this	O
pathway	O
in	O
melanocytic	B-Cell
development	O
and	O
melanoma	B-Cancer
.	O

Targeting	O
PIM	O
kinases	O
impairs	O
survival	O
of	O
hematopoietic	B-Cell
cells	I-Cell
transformed	O
by	O
kinase	O
inhibitor	O
-	O
sensitive	O
and	O
kinase	O
inhibitor	O
-	O
resistant	O
forms	O
of	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
3	I-Gene_or_gene_product
and	O
BCR	B-Gene_or_gene_product
/	O
ABL	B-Gene_or_gene_product
.	O

Previous	O
studies	O
have	O
shown	O
that	O
activation	O
of	O
the	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
5	I-Gene_or_gene_product
(	O
STAT5	B-Gene_or_gene_product
)	O
plays	O
an	O
essential	O
role	O
in	O
leukemogenesis	O
mediated	O
through	O
constitutive	O
activated	O
protein	O
tyrosine	B-Amino_acid
kinases	O
(	O
PTK	O
)	O
.	O

Because	O
PIM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
is	O
a	O
STAT5	B-Gene_or_gene_product
target	O
gene	O
,	O
we	O
analyzed	O
the	O
role	O
of	O
the	O
family	O
of	O
PIM	O
serine	B-Amino_acid
/	O
threonine	B-Amino_acid
kinases	O
(	O
PIM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
to	O
PIM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
)	O
in	O
PTK	B-Gene_or_gene_product
-	O
mediated	O
transformation	O
of	O
hematopoietic	B-Cell
cells	I-Cell
.	O

Ba	B-Cell
/	I-Cell
F3	I-Cell
cells	I-Cell
transformed	O
to	O
growth	O
factor	O
independence	O
by	O
various	O
oncogenic	O
PTKs	O
(	O
TEL	B-Gene_or_gene_product
/	O
JAK2	B-Gene_or_gene_product
,	O
TEL	B-Gene_or_gene_product
/	O
TRKC	B-Gene_or_gene_product
,	O
TEL	B-Gene_or_gene_product
/	O
ABL	B-Gene_or_gene_product
,	O
BCR	B-Gene_or_gene_product
/	O
ABL	B-Gene_or_gene_product
,	O
FLT3	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ITD	I-Gene_or_gene_product
,	O
and	O
H4	B-Gene_or_gene_product
/	O
PDGFbetaR	B-Gene_or_gene_product
)	O
show	O
abundant	O
expression	O
of	O
PIM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
PIM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
.	O

Suppression	O
of	O
PIM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
activity	O
had	O
a	O
negligible	O
effect	O
on	O
transformation	O
.	O

In	O
contrast	O
,	O
expression	O
of	O
kinase	O
-	O
dead	O
PIM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
mutant	O
(	O
PIM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2KD	I-Gene_or_gene_product
)	O
led	O
to	O
a	O
rapid	O
decline	O
of	O
survival	O
in	O
Ba	B-Cell
/	I-Cell
F3	I-Cell
cells	I-Cell
transformed	O
by	O
FLT3	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ITD	I-Gene_or_gene_product
but	O
not	O
by	O
other	O
oncogenic	O
PTKs	B-Gene_or_gene_product
tested	O
.	O

Coexpression	O
of	O
PIM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1KD	I-Gene_or_gene_product
and	O
PIM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2KD	I-Gene_or_gene_product
abrogated	O
growth	O
factor	O
-	O
independent	O
growth	O
of	O
Ba	B-Cell
/	I-Cell
F3	I-Cell
transformed	O
by	O
several	O
PTKs	O
,	O
including	O
BCR	B-Gene_or_gene_product
/	O
ABL	B-Gene_or_gene_product
.	O

Targeted	O
down	O
-	O
regulation	O
of	O
PIM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
by	O
RNA	O
interference	O
(	O
RNAi	O
)	O
selectively	O
abrogated	O
survival	O
of	O
Ba	B-Cell
/	I-Cell
F3	I-Cell
cells	I-Cell
transformed	O
by	O
various	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
3	I-Gene_or_gene_product
(	O
FLT3	B-Gene_or_gene_product
)	O
-	O
activating	O
mutants	O
[	O
internal	O
tandem	O
duplication	O
(	O
ITD	O
)	O
and	O
kinase	O
domain	O
]	O
and	O
attenuated	O
growth	O
of	O
human	B-Organism
cell	B-Cell
lines	I-Cell
containing	O
FLT3	B-Gene_or_gene_product
mutations	O
.	O

Interestingly	O
,	O
cells	B-Cell
transformed	O
by	O
FLT3	B-Gene_or_gene_product
and	O
BCR	B-Gene_or_gene_product
/	O
ABL	B-Gene_or_gene_product
mutations	O
that	O
confer	O
resistance	O
to	O
small	B-Gene_or_gene_product
-	I-Gene_or_gene_product
molecule	I-Gene_or_gene_product
tyrosine	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
inhibitors	O
were	O
still	O
sensitive	O
to	O
knockdown	O
of	O
PIM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
or	O
PIM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
PIM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
by	O
RNAi	O
.	O

Our	O
observations	O
indicate	O
that	O
combined	O
inactivation	O
of	O
PIM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
PIM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
interferes	O
with	O
oncogenic	O
PTKs	O
and	O
suggest	O
that	O
PIMs	O
are	O
alternative	O
therapeutic	O
targets	O
in	O
PTK	O
-	O
mediated	O
leukemia	B-Cancer
.	O

Targeting	O
the	O
PIM	O
kinase	O
family	O
could	O
provide	O
a	O
new	O
avenue	O
to	O
overcome	O
resistance	O
against	O
small	O
-	O
molecule	O
tyrosine	B-Amino_acid
kinase	O
inhibitors	O
.	O

Ectopic	O
localization	O
of	O
mitochondrial	B-Cellular_component
ATP	B-Gene_or_gene_product
synthase	I-Gene_or_gene_product
:	O
a	O
target	O
for	O
anti	O
-	O
angiogenesis	O
intervention	O
?	O

A	O
receptor	O
for	O
angiostatin	B-Gene_or_gene_product
was	O
identified	O
on	O
the	O
surface	O
of	O
endothelial	B-Cell
cells	I-Cell
as	O
F	B-Gene_or_gene_product
(	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	I-Gene_or_gene_product
F	I-Gene_or_gene_product
(	I-Gene_or_gene_product
0	I-Gene_or_gene_product
)	I-Gene_or_gene_product
ATP	I-Gene_or_gene_product
synthase	I-Gene_or_gene_product
(	O
Moser	O
et	O
al	O
.	O
,	O
1999	O
)	O
.	O

Proc	O
.	O

Natl	O
.	O

Acad	O
.	O

Sci	O
.	O

U	O
.	O
S	O
.	O
A	O
.	O

96	O
,	O
2811	O
-	O
2816	O
.	O

This	O
ectopic	O
ATP	B-Gene_or_gene_product
synthase	I-Gene_or_gene_product
catalyzes	O
ATP	B-Simple_chemical
synthesis	O
and	O
is	O
inhibited	O
by	O
angiostatin	B-Gene_or_gene_product
over	O
a	O
wide	O
pH	O
range	O
.	O

Endothelial	B-Cell
cells	I-Cell
grown	O
at	O
normal	O
pH	O
suffer	O
no	O
ill	O
effects	O
from	O
this	O
angiostatin	B-Gene_or_gene_product
-	O
mediated	O
inhibition	O
of	O
ATP	B-Gene_or_gene_product
synthase	I-Gene_or_gene_product
,	O
whereas	O
endothelial	B-Cell
cells	I-Cell
grown	O
at	O
low	O
,	O
tumor	B-Cancer
-	O
like	O
extracellular	B-Immaterial_anatomical_entity
pH	O
cannot	O
maintain	O
a	O
normal	O
intracellular	B-Immaterial_anatomical_entity
pH	O
and	O
die	O
.	O

Angiostatin	B-Gene_or_gene_product
inhibits	O
both	O
ATP	B-Simple_chemical
synthesis	O
and	O
ATP	B-Simple_chemical
hydrolysis	O
(	O
Moser	O
et	O
al	O
.	O
,	O
2001	O
)	O
and	O
interferes	O
with	O
intracellular	B-Immaterial_anatomical_entity
pH	O
regulation	O
(	O
Wahl	O
and	O
Grant	O
,	O
2002	O
;	O
Wahl	O
et	O
al	O
.	O
,	O
2002	O
)	O
.	O

Although	O
angiostatin	B-Gene_or_gene_product
administered	O
intravenously	B-Immaterial_anatomical_entity
is	O
cleared	O
from	O
the	O
circulation	O
in	O
a	O
matter	O
of	O
minutes	O
,	O
angiostatin	B-Simple_chemical
-	I-Simple_chemical
mimetics	I-Simple_chemical
that	O
are	O
more	O
stable	O
have	O
potential	O
for	O
clinical	O
application	O
.	O

An	O
angiostatin	B-Simple_chemical
-	I-Simple_chemical
mimetic	I-Simple_chemical
activity	O
has	O
recently	O
been	O
observed	O
using	O
a	O
polyclonal	O
antibody	O
against	O
the	O
beta	O
catalytic	O
subunit	O
of	O
ATP	B-Gene_or_gene_product
synthase	I-Gene_or_gene_product
.	O

In	O
order	O
to	O
explore	O
the	O
mechanism	O
of	O
action	O
of	O
angiostatin	B-Gene_or_gene_product
and	O
its	O
mimetics	O
,	O
further	O
work	O
needs	O
to	O
be	O
done	O
to	O
evaluate	O
clinical	O
applicability	O
,	O
specificity	O
,	O
and	O
contraindications	O
for	O
this	O
class	O
of	O
therapeutics	O
.	O

Hepatitis	B-Organism
B	I-Organism
virus	I-Organism
integration	O
event	O
in	O
human	B-Organism
chromosome	B-Cellular_component
17p	I-Cellular_component
near	O
the	O
p53	B-Gene_or_gene_product
gene	O
identifies	O
the	O
region	O
of	O
the	O
chromosome	B-Cellular_component
commonly	O
deleted	O
in	O
virus	B-Cancer
-	I-Cancer
positive	I-Cancer
hepatocellular	I-Cancer
carcinomas	I-Cancer
.	O

The	O
development	O
of	O
hepatocellular	B-Cancer
carcinoma	I-Cancer
(	O
HCC	B-Cancer
)	O
presumably	O
occurs	O
in	O
multiple	O
steps	O
and	O
is	O
influenced	O
by	O
numerous	O
factors	O
.	O

Hepatitis	B-Organism
B	I-Organism
virus	I-Organism
(	O
HBV	B-Organism
)	O
is	O
strongly	O
associated	O
with	O
the	O
development	O
of	O
HCC	B-Cancer
in	O
people	B-Organism
chronically	O
infected	O
with	O
the	O
virus	B-Organism
,	O
but	O
the	O
mechanism	O
of	O
viral	B-Organism
involvement	O
remains	O
unclear	O
.	O

One	O
possibility	O
is	O
that	O
the	O
gross	O
chromosomal	B-Cellular_component
alterations	O
frequently	O
observed	O
in	O
HCC	B-Cancer
DNA	B-Cellular_component
at	O
the	O
site	O
of	O
HBV	B-Organism
integration	O
may	O
alter	O
the	O
expression	O
of	O
important	O
nearby	O
cellular	B-Cell
genes	O
.	O

We	O
previously	O
reported	O
the	O
cloning	O
and	O
characterization	O
of	O
a	O
HBV	B-Organism
insert	O
from	O
a	O
Chinese	B-Organism
HCC	B-Cancer
.	O

The	O
viral	B-Organism
insert	O
mapped	O
to	O
chromosome	B-Cellular_component
17p11	I-Cellular_component
.	I-Cellular_component
2	I-Cellular_component
-	I-Cellular_component
12	I-Cellular_component
,	O
and	O
cellular	B-Cell
sequences	O
were	O
duplicated	O
at	O
the	O
site	O
of	O
viral	B-Organism
integration	O
.	O

In	O
the	O
present	O
study	O
a	O
DNA	B-Cellular_component
probe	O
derived	O
from	O
cellular	B-Cell
DNA	B-Cellular_component
sequences	O
adjacent	O
to	O
the	O
previously	O
characterized	O
HBV	B-Organism
insert	O
was	O
used	O
to	O
analyze	O
a	O
set	O
of	O
19	O
matched	O
normal	O
liver	B-Organ
and	O
HBV	B-Organism
-	O
positive	O
hepatoma	B-Tissue
samples	I-Tissue
obtained	O
from	O
the	O
same	O
region	O
of	O
China	O
,	O
near	O
Shanghai	O
.	O

Tumor	B-Cancer
-	O
specific	O
DNA	B-Cellular_component
changes	O
were	O
detected	O
in	O
two	O
additional	O
HCCs	B-Cancer
,	O
suggesting	O
that	O
the	O
small	O
region	O
of	O
chromosome	B-Cellular_component
17p	I-Cellular_component
defined	O
by	O
the	O
flanking	O
cell	B-Cell
DNA	B-Cellular_component
probe	O
is	O
commonly	O
altered	O
in	O
hepatomas	B-Cancer
.	O

Restriction	O
fragment	O
length	O
polymorphism	O
studies	O
demonstrated	O
that	O
the	O
loss	O
of	O
one	O
copy	O
of	O
portions	O
of	O
chromosome	B-Cellular_component
17	I-Cellular_component
occurred	O
in	O
10	O
(	O
53	O
%	O
)	O
of	O
the	O
19	O
patients	B-Organism
.	O

The	O
loss	O
of	O
one	O
allele	O
of	O
the	O
p53	B-Gene_or_gene_product
gene	O
(	O
located	O
on	O
chromosome	B-Cellular_component
17p13	I-Cellular_component
)	O
occurred	O
in	O
at	O
least	O
6	O
(	O
60	O
%	O
)	O
of	O
the	O
10	O
patients	B-Organism
who	O
were	O
heterozygous	O
at	O
the	O
p53	B-Gene_or_gene_product
locus	O
.	O

As	O
the	O
p53	B-Gene_or_gene_product
gene	O
is	O
known	O
to	O
possess	O
tumor	B-Cancer
suppressor	O
activity	O
,	O
the	O
functional	O
loss	O
of	O
this	O
gene	O
may	O
be	O
a	O
significant	O
step	O
in	O
the	O
development	O
of	O
a	O
subset	O
of	O
HCCs	B-Cancer
.	O

High	O
levels	O
of	O
allele	O
loss	O
also	O
were	O
detected	O
for	O
chromosomes	B-Cellular_component
8q	I-Cellular_component
(	O
4	O
of	O
9	O
;	O
44	O
%	O
)	O
and	O
16p	B-Cellular_component
(	O
5	O
of	O
6	O
;	O
83	O
%	O
)	O
and	O
may	O
indicate	O
the	O
presence	O
of	O
additional	O
cellular	B-Cell
genes	O
whose	O
functional	O
loss	O
is	O
important	O
in	O
the	O
development	O
of	O
HCCs	B-Cancer
.	O

Prostate	B-Gene_or_gene_product
-	I-Gene_or_gene_product
specific	I-Gene_or_gene_product
membrane	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
regulates	O
angiogenesis	O
by	O
modulating	O
integrin	B-Gene_or_gene_product
signal	O
transduction	O
.	O

The	O
transmembrane	B-Cellular_component
peptidase	O
prostate	B-Gene_or_gene_product
-	I-Gene_or_gene_product
specific	I-Gene_or_gene_product
membrane	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
(	O
PSMA	B-Gene_or_gene_product
)	O
is	O
universally	O
upregulated	O
in	O
the	O
vasculature	B-Multi-tissue_structure
of	O
solid	B-Cancer
tumors	I-Cancer
,	O
but	O
its	O
functional	O
role	O
in	O
tumor	B-Cancer
angiogenesis	O
has	O
not	O
been	O
investigated	O
.	O

Here	O
we	O
show	O
that	O
angiogenesis	O
is	O
severely	O
impaired	O
in	O
PSMA	B-Gene_or_gene_product
-	O
null	O
animals	O
and	O
that	O
this	O
angiogenic	O
defect	O
occurs	O
at	O
the	O
level	O
of	O
endothelial	B-Cell
cell	I-Cell
invasion	O
through	O
the	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
barrier	I-Cellular_component
.	O

Because	O
proteolytic	O
degradation	O
of	O
the	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
is	O
a	O
critical	O
component	O
of	O
endothelial	B-Cell
invasion	O
in	O
angiogenesis	O
,	O
it	O
is	O
logical	O
to	O
assume	O
that	O
PSMA	B-Gene_or_gene_product
participates	O
in	O
matrix	B-Cellular_component
degradation	O
.	O

However	O
,	O
we	O
demonstrate	O
a	O
novel	O
and	O
more	O
complex	O
role	O
for	O
PSMA	B-Gene_or_gene_product
in	O
angiogenesis	O
,	O
where	O
it	O
is	O
a	O
principal	O
component	O
of	O
a	O
regulatory	O
loop	O
that	O
is	O
tightly	O
modulating	O
laminin	B-Gene_or_gene_product
-	O
specific	O
integrin	B-Gene_or_gene_product
signaling	O
and	O
GTPase	O
-	O
dependent	O
,	O
p21	B-Gene_or_gene_product
-	I-Gene_or_gene_product
activated	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
PAK	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
activity	O
.	O

We	O
show	O
that	O
PSMA	B-Gene_or_gene_product
inhibition	O
,	O
knockdown	O
,	O
or	O
deficiency	O
decreases	O
endothelial	B-Cell
cell	I-Cell
invasion	O
in	O
vitro	O
via	O
integrin	B-Gene_or_gene_product
and	O
PAK	B-Gene_or_gene_product
,	O
thus	O
abrogating	O
angiogenesis	O
.	O

Interestingly	O
,	O
the	O
neutralization	O
of	O
beta	B-Gene_or_gene_product
(	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	I-Gene_or_gene_product
or	O
the	O
inactivation	O
of	O
PAK	B-Gene_or_gene_product
increases	O
PSMA	B-Gene_or_gene_product
activity	O
,	O
suggesting	O
that	O
they	O
negatively	O
regulate	O
PSMA	B-Gene_or_gene_product
.	O

This	O
negative	O
regulation	O
is	O
mediated	O
by	O
the	O
cytoskeleton	B-Cellular_component
as	O
the	O
disruption	O
of	O
interactions	O
between	O
the	O
PSMA	B-Gene_or_gene_product
cytoplasmic	B-Organism_substance
tail	O
and	O
the	O
anchor	O
protein	O
filamin	B-Gene_or_gene_product
A	I-Gene_or_gene_product
decreases	O
PSMA	B-Gene_or_gene_product
activity	O
,	O
integrin	B-Gene_or_gene_product
function	O
,	O
and	O
PAK	B-Gene_or_gene_product
activation	O
.	O

Finally	O
,	O
the	O
inhibition	O
of	O
PAK	B-Gene_or_gene_product
activation	O
enhances	O
the	O
PSMA	B-Gene_or_gene_product
/	O
filamin	B-Gene_or_gene_product
A	I-Gene_or_gene_product
interaction	O
and	O
,	O
thus	O
,	O
boosts	O
PSMA	B-Gene_or_gene_product
activity	O
.	O

These	O
data	O
imply	O
that	O
PSMA	B-Gene_or_gene_product
participates	O
in	O
an	O
autoregulatory	O
loop	O
,	O
wherein	O
active	O
PSMA	B-Gene_or_gene_product
facilitates	O
integrin	B-Gene_or_gene_product
signaling	O
and	O
PAK	B-Gene_or_gene_product
activation	O
,	O
leading	O
to	O
both	O
productive	O
invasion	O
and	O
downregulation	O
of	O
integrin	B-Gene_or_gene_product
beta	I-Gene_or_gene_product
(	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	I-Gene_or_gene_product
signaling	O
via	O
reduced	O
PSMA	B-Gene_or_gene_product
activity	O
.	O

Therefore	O
,	O
we	O
have	O
identified	O
a	O
novel	O
role	O
for	O
PSMA	B-Gene_or_gene_product
as	O
a	O
true	O
molecular	O
interface	O
,	O
integrating	O
both	O
extracellular	B-Immaterial_anatomical_entity
and	O
intracellular	B-Immaterial_anatomical_entity
signals	O
during	O
angiogenesis	O
.	O

Heparin	B-Simple_chemical
immobilized	O
porous	O
PLGA	B-Simple_chemical
microspheres	O
for	O
angiogenic	O
growth	O
factor	O
delivery	O
.	O

PURPOSE	O
:	O
Heparin	B-Simple_chemical
immobilized	O
porous	O
poly	B-Simple_chemical
(	I-Simple_chemical
D	I-Simple_chemical
,	I-Simple_chemical
L	I-Simple_chemical
-	I-Simple_chemical
lactic	I-Simple_chemical
-	I-Simple_chemical
co	I-Simple_chemical
-	I-Simple_chemical
glycolic	I-Simple_chemical
acid	I-Simple_chemical
)	I-Simple_chemical
(	O
PLGA	B-Simple_chemical
)	O
microspheres	O
were	O
prepared	O
for	O
sustained	O
release	O
of	O
basic	B-Gene_or_gene_product
fibroblast	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
bFGF	B-Gene_or_gene_product
)	O
to	O
induce	O
angiogenesis	O
.	O

MATERIALS	O
AND	O
METHODS	O
:	O
Porous	O
PLGA	B-Simple_chemical
microspheres	O
having	O
primary	O
amine	O
groups	O
on	O
the	O
surface	O
were	O
prepared	O
using	O
an	O
oil	O
-	O
in	O
-	O
water	O
(	O
O	O
/	O
W	O
)	O
single	O
emulsion	O
method	O
using	O
Pluronic	O
F	O
-	O
127	O
as	O
an	O
extractable	O
porogen	O
.	O

Heparin	B-Simple_chemical
was	O
surface	O
immobilized	O
via	O
covalent	O
conjugation	O
.	O

bFGF	B-Gene_or_gene_product
was	O
loaded	O
into	O
the	O
heparin	B-Simple_chemical
functionalized	O
(	O
PLGA	B-Simple_chemical
-	O
heparin	B-Simple_chemical
)	O
microspheres	O
by	O
a	O
simple	O
dipping	O
method	O
.	O

The	O
bFGF	B-Gene_or_gene_product
loaded	O
PLGA	B-Simple_chemical
-	O
heparin	B-Simple_chemical
microspheres	O
were	O
tested	O
for	O
in	O
vitro	O
release	O
and	O
in	O
vivo	O
angiogenic	O
activity	O
.	O

RESULTS	O
:	O
PLGA	B-Simple_chemical
microspheres	O
with	O
an	O
open	O
-	O
porous	O
structure	O
were	O
formed	O
.	O

The	O
amount	O
of	O
conjugated	O
amine	O
group	O
onto	O
the	O
microspheres	O
was	O
1	O
.	O
93	O
+	O
/	O
-	O
0	O
.	O
01	O
nmol	O
/	O
mg	O
-	O
microspheres	O
,	O
while	O
the	O
amount	O
of	O
heparin	B-Simple_chemical
was	O
95	O
.	O
8	O
pmol	O
/	O
mg	O
-	O
microspheres	O
.	O

PLGA	B-Simple_chemical
-	O
heparin	B-Simple_chemical
microspheres	O
released	O
out	O
bFGF	B-Gene_or_gene_product
in	O
a	O
more	O
sustained	O
manner	O
with	O
a	O
smaller	O
extent	O
of	O
initial	O
burst	O
than	O
PLGA	B-Simple_chemical
microspheres	O
,	O
indicating	O
that	O
surface	O
immobilized	O
heparin	B-Simple_chemical
controlled	O
the	O
release	O
rate	O
of	O
bFGF	B-Gene_or_gene_product
.	O

Subcutaneous	O
implantation	O
of	O
bFGF	B-Gene_or_gene_product
loaded	O
PLGA	B-Simple_chemical
-	O
heparin	B-Simple_chemical
microspheres	O
in	O
mice	B-Organism
significantly	O
induced	O
the	O
formation	O
of	O
new	O
vascular	B-Tissue
microvessels	I-Tissue
.	O

CONCLUSIONS	O
:	O
PLGA	B-Simple_chemical
microspheres	O
with	O
an	O
open	O
porous	O
structure	O
allowed	O
significant	O
amount	O
of	O
heparin	B-Simple_chemical
immobilization	O
and	O
bFGF	B-Gene_or_gene_product
loading	O
.	O

bFGF	B-Gene_or_gene_product
loaded	O
PLGA	B-Simple_chemical
-	O
HP	B-Simple_chemical
microspheres	O
showed	O
sustained	O
release	O
profiles	O
of	O
bFGF	B-Gene_or_gene_product
in	O
vitro	O
,	O
demonstrating	O
reversible	O
and	O
specific	O
binding	O
of	O
bFGF	B-Gene_or_gene_product
to	O
immobilized	O
heparin	B-Simple_chemical
.	O

They	O
also	O
induced	O
local	O
angiogenesis	O
in	O
vivo	O
in	O
an	O
animal	O
model	O
.	O

The	O
role	O
of	O
syndecans	B-Gene_or_gene_product
in	O
disease	O
and	O
wound	B-Pathological_formation
healing	O
.	O

Syndecans	B-Gene_or_gene_product
are	O
a	O
family	O
of	O
transmembrane	B-Cellular_component
heparan	B-Simple_chemical
sulfate	I-Simple_chemical
proteoglycans	O
widely	O
expressed	O
in	O
both	O
developing	B-Tissue
and	O
adult	B-Tissue
tissues	I-Tissue
.	O

Until	O
recently	O
,	O
their	O
role	O
in	O
pathogenesis	O
was	O
largely	O
unexplored	O
.	O

In	O
this	O
review	O
,	O
we	O
discuss	O
the	O
reported	O
involvement	O
of	O
syndecans	B-Gene_or_gene_product
in	O
human	B-Organism
cancers	B-Cancer
,	O
infectious	O
diseases	O
,	O
obesity	O
,	O
wound	B-Pathological_formation
healing	O
and	O
angiogenesis	O
.	O

In	O
some	O
cancers	B-Cancer
,	O
syndecan	B-Gene_or_gene_product
expression	O
has	O
been	O
shown	O
to	O
regulate	O
tumor	B-Cell
cell	I-Cell
function	O
(	O
e	O
.	O
g	O
.	O
proliferation	O
,	O
adhesion	O
,	O
and	O
motility	O
)	O
and	O
serve	O
as	O
a	O
prognostic	O
marker	O
for	O
tumor	B-Cancer
progression	O
and	O
patient	B-Organism
survival	O
.	O

The	O
ectodomains	O
and	O
heparan	B-Simple_chemical
sulfate	I-Simple_chemical
glycosaminoglycan	I-Simple_chemical
chains	O
of	O
syndecans	B-Gene_or_gene_product
can	O
also	O
act	O
as	O
receptors	O
/	O
co	O
-	O
receptors	O
for	O
some	O
bacterial	O
and	O
viral	O
pathogens	O
,	O
mediating	O
infection	O
.	O

In	O
addition	O
,	O
syndecans	B-Gene_or_gene_product
bind	O
to	O
obesity	O
-	O
related	O
factors	O
and	O
regulate	O
their	O
signaling	O
,	O
in	O
turn	O
modulating	O
food	O
consumption	O
and	O
weight	O
balance	O
.	O

In	O
vivo	O
animal	O
models	O
of	O
tissue	B-Tissue
injury	O
and	O
in	O
vitro	O
data	O
also	O
implicate	O
syndecans	B-Gene_or_gene_product
in	O
processes	O
necessary	O
for	O
wound	B-Pathological_formation
healing	O
,	O
including	O
fibroblast	B-Cell
and	O
endothelial	B-Cell
proliferation	O
,	O
cell	B-Cell
motility	O
,	O
angiogenesis	O
,	O
and	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
organization	O
.	O

These	O
new	O
insights	O
into	O
the	O
involvement	O
of	O
syndecans	B-Gene_or_gene_product
in	O
disease	O
and	O
tissue	B-Tissue
repair	O
coupled	O
with	O
the	O
emergence	O
of	O
syndecan	B-Gene_or_gene_product
-	O
specific	O
molecular	O
tools	O
may	O
lead	O
to	O
novel	O
therapies	O
for	O
a	O
variety	O
of	O
human	B-Organism
diseases	O
.	O

C	B-Gene_or_gene_product
/	I-Gene_or_gene_product
EBPbeta	I-Gene_or_gene_product
is	O
over	O
-	O
expressed	O
in	O
gastric	B-Organ
carcinogenesis	O
and	O
is	O
associated	O
with	O
COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
expression	O
.	O

The	O
CCAAT	B-Gene_or_gene_product
/	I-Gene_or_gene_product
enhancer	I-Gene_or_gene_product
-	I-Gene_or_gene_product
binding	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
(	O
C	B-Gene_or_gene_product
/	I-Gene_or_gene_product
EBPbeta	I-Gene_or_gene_product
)	O
transcription	O
factor	O
has	O
been	O
associated	O
with	O
several	O
cancer	B-Cancer
models	O
.	O

In	O
this	O
study	O
,	O
the	O
expression	O
of	O
C	B-Gene_or_gene_product
/	I-Gene_or_gene_product
EBPbeta	I-Gene_or_gene_product
was	O
analysed	O
in	O
a	O
series	O
of	O
90	O
gastric	B-Cancer
carcinomas	I-Cancer
(	O
GCs	B-Cancer
)	O
.	O

We	O
also	O
assessed	O
the	O
effect	O
of	O
C	B-Gene_or_gene_product
/	I-Gene_or_gene_product
EBPbeta	I-Gene_or_gene_product
on	O
COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
expression	O
.	O

In	O
normal	O
gastric	B-Multi-tissue_structure
mucosa	I-Multi-tissue_structure
,	O
C	B-Gene_or_gene_product
/	I-Gene_or_gene_product
EBPbeta	I-Gene_or_gene_product
expression	O
was	O
restricted	O
to	O
cells	B-Cell
in	O
the	O
proliferative	O
zone	B-Immaterial_anatomical_entity
.	O

In	O
intestinal	B-Pathological_formation
metaplasia	I-Pathological_formation
,	O
dysplasia	B-Pathological_formation
,	O
and	O
GC	B-Cancer
of	O
the	O
intestinal	B-Multi-tissue_structure
and	O
atypical	O
subtypes	O
,	O
C	B-Gene_or_gene_product
/	I-Gene_or_gene_product
EBPbeta	I-Gene_or_gene_product
was	O
over	O
-	O
expressed	O
(	O
p	O
<	O
0	O
.	O
0001	O
,	O
for	O
the	O
association	O
with	O
histological	O
type	O
)	O
.	O

C	B-Gene_or_gene_product
/	I-Gene_or_gene_product
EBPbeta	I-Gene_or_gene_product
and	O
Ki67	B-Gene_or_gene_product
,	O
a	O
marker	O
of	O
cell	B-Cell
proliferation	O
,	O
were	O
also	O
co	O
-	O
expressed	O
in	O
primary	O
GC	B-Cancer
.	O

We	O
also	O
observed	O
an	O
overlap	O
between	O
C	B-Gene_or_gene_product
/	I-Gene_or_gene_product
EBPbeta	I-Gene_or_gene_product
and	O
COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
expression	O
in	O
GC	B-Cancer
.	O

Using	O
GC	B-Cell
cell	I-Cell
lines	I-Cell
we	O
show	O
that	O
C	B-Gene_or_gene_product
/	I-Gene_or_gene_product
EBPbeta	I-Gene_or_gene_product
can	O
regulate	O
the	O
expression	O
of	O
endogenous	O
COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
transactivate	O
the	O
promoter	O
of	O
the	O
COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
gene	O
,	O
depending	O
on	O
its	O
methylation	O
status	O
.	O

These	O
results	O
suggest	O
that	O
C	B-Gene_or_gene_product
/	I-Gene_or_gene_product
EBPbeta	I-Gene_or_gene_product
may	O
be	O
a	O
marker	O
of	O
neoplastic	O
transformation	O
and	O
also	O
play	O
an	O
active	O
role	O
in	O
gastric	B-Organ
tumourigenesis	O
by	O
regulating	O
COX	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
expression	O
.	O

Lysophosphatidic	B-Simple_chemical
acid	I-Simple_chemical
downregulates	O
tissue	B-Gene_or_gene_product
inhibitor	I-Gene_or_gene_product
of	I-Gene_or_gene_product
metalloproteinases	I-Gene_or_gene_product
,	O
which	O
are	O
negatively	O
involved	O
in	O
lysophosphatidic	B-Simple_chemical
acid	I-Simple_chemical
-	O
induced	O
cell	B-Cell
invasion	O
.	O

Ovarian	B-Cancer
cancer	I-Cancer
is	O
a	O
highly	O
metastatic	O
disease	O
.	O

Lysophosphatidic	B-Simple_chemical
acid	I-Simple_chemical
(	O
LPA	B-Simple_chemical
)	O
levels	O
are	O
elevated	O
in	O
ascites	B-Organism_substance
from	O
ovarian	B-Cancer
cancer	I-Cancer
patients	B-Organism
,	O
but	O
its	O
potential	O
role	O
in	O
ovarian	B-Cancer
cancer	I-Cancer
metastasis	O
has	O
just	O
begun	O
to	O
be	O
revealed	O
.	O

In	O
this	O
work	O
,	O
we	O
show	O
that	O
LPA	B-Simple_chemical
stimulates	O
invasion	O
of	O
primary	O
ovarian	B-Cell
cancer	I-Cell
cells	I-Cell
,	O
but	O
not	O
ovarian	B-Cell
epithelial	I-Cell
or	O
borderline	B-Cell
ovarian	I-Cell
tumor	I-Cell
cells	I-Cell
,	O
although	O
these	O
benign	B-Cell
cells	I-Cell
indeed	O
respond	O
to	O
LPA	B-Simple_chemical
in	O
cell	B-Cell
migration	O
.	O

We	O
have	O
found	O
that	O
LPA	B-Simple_chemical
downregulates	O
tissue	B-Gene_or_gene_product
inhibitor	I-Gene_or_gene_product
of	I-Gene_or_gene_product
metalloproteinases	I-Gene_or_gene_product
(	O
TIMPs	B-Gene_or_gene_product
)	O
.	O

TIMP2	B-Gene_or_gene_product
and	O
TIMP3	B-Gene_or_gene_product
play	O
functional	O
role	O
in	O
LPA	B-Simple_chemical
-	O
induced	O
invasion	O
as	O
negative	O
regulators	O
.	O

G	B-Gene_or_gene_product
(	I-Gene_or_gene_product
i	I-Gene_or_gene_product
)	I-Gene_or_gene_product
protein	O
,	O
phosphatidylinositol	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
(	O
PI3K	B-Gene_or_gene_product
)	O
,	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
,	O
cytosolic	B-Organism_substance
phospholipase	B-Gene_or_gene_product
A	I-Gene_or_gene_product
(	I-Gene_or_gene_product
2	I-Gene_or_gene_product
)	I-Gene_or_gene_product
and	O
urokinase	B-Gene_or_gene_product
type	I-Gene_or_gene_product
plasminogen	I-Gene_or_gene_product
activator	I-Gene_or_gene_product
(	O
uPA	B-Gene_or_gene_product
)	O
are	O
required	O
for	O
LPA	B-Simple_chemical
-	O
induced	O
cells	B-Cell
invasion	O
.	O

TIMP3	B-Gene_or_gene_product
may	O
affect	O
two	O
independent	O
downstream	O
targets	O
,	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
and	O
p38	B-Gene_or_gene_product
MAPK	I-Gene_or_gene_product
.	O

In	O
vivo	O
,	O
LPA	B-Simple_chemical
stimulates	O
tumor	B-Cancer
metastasis	O
in	O
an	O
orthotopic	O
ovarian	B-Cancer
tumor	I-Cancer
model	O
,	O
which	O
can	O
be	O
inhibited	O
by	O
a	O
PI3K	B-Gene_or_gene_product
inhibitor	O
,	O
LY294002	B-Simple_chemical
.	O

In	O
summary	O
,	O
LPA	B-Simple_chemical
is	O
likely	O
a	O
key	O
component	O
for	O
promoting	O
ovarian	B-Organ
metastasis	O
in	O
vivo	O
.	O

LPA	B-Simple_chemical
downregulates	O
TIMP3	B-Gene_or_gene_product
,	O
which	O
may	O
have	O
targets	O
other	O
than	O
metalloproteinases	O
.	O

Our	O
in	O
vivo	O
metastasis	O
mouse	B-Organism
model	O
is	O
useful	O
for	O
studying	O
the	O
efficacy	O
of	O
therapeutic	O
regimes	O
of	O
ovarian	B-Cancer
cancer	I-Cancer
.	O

Role	O
of	O
the	O
fibrinolytic	O
and	O
matrix	B-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
systems	O
in	O
development	O
of	O
adipose	B-Tissue
tissue	I-Tissue
.	O

Obesity	O
is	O
a	O
common	O
disorder	O
and	O
related	O
diseases	O
such	O
as	O
diabetes	O
,	O
atherosclerosis	O
,	O
hypertension	O
,	O
cardiovascular	B-Anatomical_system
disease	O
and	O
cancer	B-Cancer
are	O
a	O
major	O
cause	O
of	O
mortality	O
and	O
morbidity	O
in	O
Western	O
-	O
type	O
societies	O
.	O

Development	O
of	O
obesity	O
is	O
associated	O
with	O
extensive	O
modifications	O
in	O
adipose	B-Tissue
tissue	I-Tissue
involving	O
adipogenesis	O
,	O
angiogenesis	O
and	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
proteolysis	O
.	O

The	O
fibrinolytic	O
(	O
plasminogen	B-Gene_or_gene_product
/	O
plasmin	B-Gene_or_gene_product
)	O
and	O
matrix	B-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
(	O
MMP	B-Gene_or_gene_product
)	O
systems	O
cooperate	O
in	O
these	O
processes	O
.	O

A	O
nutritionally	O
induced	O
obesity	O
model	O
in	O
transgenic	O
mice	B-Organism
has	O
been	O
used	O
extensively	O
to	O
study	O
the	O
role	O
of	O
the	O
fibrinolytic	O
and	O
MMP	B-Gene_or_gene_product
systems	O
in	O
the	O
development	O
of	O
obesity	O
.	O

These	O
studies	O
support	O
a	O
role	O
of	O
both	O
systems	O
in	O
adipogenesis	O
and	O
obesity	O
;	O
the	O
role	O
of	O
specific	O
members	O
of	O
these	O
families	O
,	O
however	O
,	O
remains	O
to	O
be	O
determined	O
.	O

Extracellular	B-Cellular_component
matrix	I-Cellular_component
as	O
a	O
bioactive	O
material	O
for	O
soft	B-Tissue
tissue	I-Tissue
reconstruction	O
.	O

The	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
(	O
ECM	B-Cellular_component
)	O
directs	O
all	O
phases	O
of	O
healing	O
following	O
trauma	O
or	O
disease	O
and	O
is	O
therefore	O
a	O
natural	O
source	O
of	O
prosthetic	O
mesh	O
material	O
that	O
can	O
be	O
used	O
strategically	O
to	O
induce	O
the	O
repair	O
and	O
restoration	O
of	O
soft	B-Tissue
tissues	I-Tissue
following	O
surgery	O
.	O

Biomaterials	O
such	O
as	O
Surgisis	O
(	O
Cook	O
Biotech	O
Incorporated	O
,	O
West	O
Lafayette	O
,	O
IN	O
,	O
USA	O
)	O
,	O
which	O
are	O
derived	O
from	O
natural	O
ECM	B-Cellular_component
,	O
provide	O
the	O
extracellular	B-Cellular_component
components	I-Cellular_component
necessary	O
to	O
direct	O
the	O
healing	O
response	O
,	O
allow	O
for	O
the	O
proliferation	O
of	O
new	O
,	O
healthy	O
tissue	B-Tissue
and	O
restore	O
tissue	B-Tissue
integrity	O
to	O
the	O
damaged	O
site	O
.	O

The	O
3	O
-	O
D	O
organization	O
of	O
these	O
extracellular	B-Cellular_component
components	I-Cellular_component
distinguishes	O
the	O
Surgisis	O
mesh	O
from	O
synthetic	O
materials	O
and	O
is	O
associated	O
with	O
constructive	O
tissue	B-Tissue
remodelling	O
instead	O
of	O
scar	B-Tissue
tissue	I-Tissue
.	O

Common	O
features	O
of	O
this	O
ECM	B-Cellular_component
-	O
assisted	O
tissue	B-Tissue
remodelling	O
include	O
angiogenesis	O
,	O
recruitment	O
of	O
circulating	O
progenitor	B-Cell
cells	I-Cell
and	O
constructive	O
remodelling	O
of	O
damaged	O
tissue	B-Tissue
structures	O
.	O

The	O
tissue	B-Tissue
response	O
to	O
this	O
biologic	O
mesh	O
is	O
discussed	O
in	O
the	O
context	O
of	O
recent	O
reports	O
on	O
clinical	O
hernia	B-Pathological_formation
repair	O
.	O

[	O
Study	O
on	O
the	O
expression	O
of	O
angiogenesis	O
and	O
spontaneous	O
apoptosis	O
and	O
their	O
relevance	O
in	O
laryngeal	B-Cancer
squamous	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
]	O

OBJECTIVE	O
:	O
To	O
investigate	O
the	O
relationship	O
among	O
angiogenesis	O
,	O
spontaneous	O
apoptosis	O
and	O
clinicopathological	O
parameters	O
in	O
laryngeal	B-Cancer
squamous	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
(	O
LSCC	B-Cancer
)	O
.	O

METHOD	O
:	O
The	O
intratumor	B-Tissue
microvessel	I-Tissue
density	O
(	O
IMVD	O
)	O
,	O
apoptotic	O
index	O
(	O
AI	O
)	O
and	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
expression	O
were	O
detected	O
by	O
immunohistochemistry	O
SABC	O
and	O
terminal	O
uridine	O
deoxynucleotidyl	O
transferase	O
mediated	O
nick	O
end	O
labeling	O
(	O
TUNEL	O
)	O
methods	O
in34	O
LSCC	B-Cancer
patients	B-Organism
.	O

RESULT	O
:	O
The	O
average	O
IMVD	O
was	O
(	O
21	O
.	O
50	O
+	O
/	O
-	O
8	O
.	O
87	O
)	O
,	O
and	O
median	O
of	O
AI	O
was	O
1	O
.	O
15	O
%	O
.	O

The	O
average	O
IMVD	O
in	O
positive	O
and	O
negative	O
cervical	B-Multi-tissue_structure
lymphatic	I-Multi-tissue_structure
metastasis	O
was	O
(	O
26	O
.	O
33	O
+	O
/	O
-	O
9	O
.	O
70	O
)	O
and	O
(	O
17	O
.	O
68	O
+	O
/	O
-	O
6	O
.	O
06	O
)	O
respectively	O
,	O
and	O
the	O
IMVD	O
with	O
positive	O
lymphatic	B-Cancer
metastasis	I-Cancer
tumors	I-Cancer
was	O
statistical	O
significantly	O
higher	O
than	O
those	O
with	O
negative	O
cervical	B-Cancer
lymphatic	I-Cancer
metastasis	I-Cancer
tumors	I-Cancer
(	O
P	O
less	O
than	O
0	O
.	O
01	O
)	O
.	O

The	O
average	O
IMVD	O
had	O
statistical	O
difference	O
in	O
histological	O
grading	O
(	O
P	O
less	O
than	O
0	O
.	O
01	O
)	O
,	O
and	O
analysis	O
by	O
one	O
to	O
one	O
,	O
the	O
average	O
IMVD	O
had	O
statistical	O
difference	O
between	O
high	O
and	O
median	O
grading	O
.	O

Expression	O
of	O
VEGF	B-Gene_or_gene_product
had	O
a	O
significantly	O
positive	O
correlation	O
with	O
IMVD	O
(	O
r	O
=	O
0	O
.	O
51	O
,	O
P	O
less	O
than	O
0	O
.	O
01	O
)	O
.	O

Statistical	O
analysis	O
revealed	O
a	O
significantly	O
inverse	O
correlation	O
between	O
AI	O
and	O
IMVD	O
(	O
r	O
=	O
-	O
0	O
.	O
53	O
,	O
P	O
less	O
than	O
0	O
.	O
01	O
)	O
.	O

We	O
failed	O
to	O
find	O
the	O
statistical	O
difference	O
between	O
IMVD	O
and	O
tumor	B-Cancer
T	O
-	O
stage	O
in	O
LSCC	B-Cancer
.	O

CONCLUSION	O
:	O
IMVD	O
may	O
be	O
an	O
important	O
indicator	O
to	O
predict	O
cervical	B-Multi-tissue_structure
lymphatic	I-Multi-tissue_structure
metastasis	O
in	O
LSCC	B-Cancer
.	O

VEGF	B-Gene_or_gene_product
might	O
be	O
an	O
important	O
angiogenic	O
factor	O
,	O
and	O
could	O
promote	O
tumor	B-Cancer
angiogenesis	O
in	O
LSCC	B-Cancer
.	O

Tumor	B-Cancer
angiogenesis	O
might	O
contribute	O
to	O
tumor	B-Cancer
malignant	O
progression	O
by	O
inhibiting	O
spontaneous	O
apoptosis	O
in	O
LSCC	B-Cancer
.	O

The	O
cancer	B-Cell
cell	I-Cell
'	O
s	O
"	O
power	O
plants	O
"	O
as	O
promising	O
therapeutic	O
targets	O
:	O
an	O
overview	O
.	O

This	O
introductory	O
article	O
to	O
the	O
review	O
series	O
entitled	O
"	O
The	O
Cancer	B-Cell
Cell	I-Cell
'	O
s	O
Power	O
Plants	O
as	O
Promising	O
Therapeutic	O
Targets	O
"	O
is	O
written	O
while	O
more	O
than	O
20	O
million	O
people	O
suffer	O
from	O
cancer	B-Cancer
.	O

It	O
summarizes	O
strategies	O
to	O
destroy	O
or	O
prevent	O
cancers	B-Cancer
by	O
targeting	O
their	O
energy	O
production	O
factories	O
,	O
i	O
.	O
e	O
.	O
,	O
"	O
power	O
plants	O
.	O
"	O
All	O
nucleated	O
animal	B-Organism
/	O
human	B-Organism
cells	B-Cell
have	O
two	O
types	O
of	O
power	O
plants	O
,	O
i	O
.	O
e	O
.	O
,	O
systems	O
that	O
make	O
the	O
"	O
high	O
energy	O
"	O
compound	O
ATP	B-Simple_chemical
from	O
ADP	B-Simple_chemical
and	O
P	B-Simple_chemical
(	I-Simple_chemical
i	I-Simple_chemical
)	I-Simple_chemical
.	O

One	O
type	O
is	O
"	O
glycolysis	O
,	O
"	O
the	O
other	O
the	O
"	O
mitochondria	B-Cellular_component
.	O
"	O
In	O
contrast	O
to	O
most	O
normal	O
cells	B-Cell
where	O
the	O
mitochondria	B-Cellular_component
are	O
the	O
major	O
ATP	B-Simple_chemical
producers	O
(	O
>	O
90	O
%	O
)	O
in	O
fueling	O
growth	O
,	O
human	B-Organism
cancers	B-Cancer
detected	O
via	O
Positron	O
Emission	O
Tomography	O
(	O
PET	O
)	O
rely	O
on	O
both	O
types	O
of	O
power	O
plants	O
.	O

In	O
such	O
cancers	B-Cancer
,	O
glycolysis	O
may	O
contribute	O
nearly	O
half	O
the	O
ATP	B-Simple_chemical
even	O
in	O
the	O
presence	O
of	O
oxygen	B-Simple_chemical
(	O
"	O
Warburg	O
effect	O
"	O
)	O
.	O

Based	O
solely	O
on	O
cell	B-Cell
energetics	O
,	O
this	O
presents	O
a	O
challenge	O
to	O
identify	O
curative	O
agents	O
that	O
destroy	O
only	O
cancer	B-Cell
cells	I-Cell
as	O
they	O
must	O
destroy	O
both	O
of	O
their	O
power	O
plants	O
causing	O
"	O
necrotic	O
cell	B-Cell
death	O
"	O
and	O
leave	O
normal	O
cells	B-Cell
alone	O
.	O

One	O
such	O
agent	O
,	O
3	B-Simple_chemical
-	I-Simple_chemical
bromopyruvate	I-Simple_chemical
(	O
3	B-Simple_chemical
-	I-Simple_chemical
BrPA	I-Simple_chemical
)	O
,	O
a	O
lactic	B-Simple_chemical
acid	I-Simple_chemical
analog	I-Simple_chemical
,	O
has	O
been	O
shown	O
to	O
inhibit	O
both	O
glycolytic	O
and	O
mitochondrial	B-Cellular_component
ATP	B-Simple_chemical
production	O
in	O
rapidly	O
growing	O
cancers	B-Cancer
(	O
Ko	O
et	O
al	O
.	O
,	O
Cancer	O
Letts	O
.	O
,	O
173	O
,	O
83	O
-	O
91	O
,	O
2001	O
)	O
,	O
leave	O
normal	O
cells	B-Cell
alone	O
,	O
and	O
eradicate	O
advanced	O
cancers	B-Cancer
(	O
19	O
of	O
19	O
)	O
in	O
a	O
rodent	B-Organism
model	O
(	O
Ko	O
et	O
al	O
.	O
,	O
Biochem	O
.	O
Biophys	O
.	O
Res	O
.	O
Commun	O
.	O
,	O
324	O
,	O
269	O
-	O
275	O
,	O
2004	O
)	O
.	O

A	O
second	O
approach	O
is	O
to	O
induce	O
only	O
cancer	B-Cell
cells	I-Cell
to	O
undergo	O
"	O
apoptotic	O
cell	B-Cell
death	O
.	O
"	O
Here	O
,	O
mitochondria	B-Cellular_component
release	O
cell	B-Cell
death	O
inducing	O
factors	O
(	O
e	O
.	O
g	O
.	O
,	O
cytochrome	B-Gene_or_gene_product
c	I-Gene_or_gene_product
)	O
.	O

In	O
a	O
third	O
approach	O
,	O
cancer	B-Cell
cells	I-Cell
are	O
induced	O
to	O
die	O
by	O
both	O
apoptotic	O
and	O
necrotic	O
events	O
.	O

In	O
summary	O
,	O
much	O
effort	O
is	O
being	O
focused	O
on	O
identifying	O
agents	O
that	O
induce	O
"	O
necrotic	O
,	O
"	O
"	O
apoptotic	O
"	O
or	O
apoptotic	O
plus	O
necrotic	O
cell	O
death	O
only	O
in	O
cancer	B-Cell
cells	I-Cell
.	O

Regardless	O
how	O
death	O
is	O
inflicted	O
,	O
every	O
cancer	B-Cell
cell	I-Cell
must	O
die	O
,	O
be	O
it	O
fast	O
or	O
slow	O
.	O

Pathological	O
animal	O
models	O
in	O
the	O
experimental	O
evaluation	O
of	O
tumour	B-Tissue
microvasculature	I-Tissue
with	O
magnetic	O
resonance	O
imaging	O
.	O

PURPOSE	O
:	O
The	O
purpose	O
of	O
this	O
study	O
was	O
to	O
evaluate	O
the	O
applications	O
of	O
magnetic	O
resonance	O
imaging	O
(	O
MRI	O
)	O
,	O
and	O
in	O
particular	O
,	O
dynamic	O
contrast	O
-	O
enhanced	O
MRI	O
(	O
DCE	O
-	O
MRI	O
)	O
,	O
in	O
the	O
assessment	O
of	O
tumour	B-Tissue
microvasculature	I-Tissue
by	O
means	O
of	O
animal	O
tumour	B-Cancer
models	O
evaluated	O
before	O
and	O
after	O
antiangiogenic	O
treatment	O
.	O

MATERIALS	O
AND	O
METHODS	O
:	O
Forty	O
-	O
two	O
MRI	O
exams	O
were	O
performed	O
with	O
intravascular	B-Immaterial_anatomical_entity
contrast	O
media	O
in	O
21	O
rats	B-Organism
:	O
tumours	B-Cancer
were	O
induced	O
by	O
subcutaneous	O
injection	O
of	O
colon	B-Cell
carcinoma	I-Cell
cells	I-Cell
in	O
7	O
rats	B-Organism
and	O
mammary	B-Cell
adenocarcinoma	I-Cell
cells	I-Cell
in	O
14	O
rats	B-Organism
.	O

Perfusion	O
and	O
permeability	O
parameters	O
of	O
the	O
implanted	O
tumours	B-Cancer
were	O
evaluated	O
by	O
using	O
two	O
contrast	O
media	O
(	O
B22956	B-Simple_chemical
/	I-Simple_chemical
1	I-Simple_chemical
and	O
Gd	B-Simple_chemical
-	I-Simple_chemical
DTPA37	I-Simple_chemical
-	I-Simple_chemical
albumin	I-Simple_chemical
)	O
to	O
establish	O
response	O
to	O
treatment	O
with	O
two	O
different	O
antiangiogenic	O
drugs	O
(	O
tamoxifen	B-Simple_chemical
and	O
SU6668	B-Simple_chemical
)	O
.	O

These	O
parameters	O
were	O
correlated	O
with	O
histology	O
to	O
obtain	O
a	O
radiological	O
-	O
histological	O
map	O
of	O
tumour	B-Tissue
microvasculature	I-Tissue
.	O

RESULTS	O
:	O
DCE	O
-	O
MRI	O
revealed	O
greater	O
enhancement	O
in	O
the	O
peripheral	B-Organism_subdivision
area	I-Organism_subdivision
than	O
in	O
the	O
central	B-Organism_subdivision
area	I-Organism_subdivision
in	O
all	O
the	O
examined	O
animal	O
models	O
.	O

In	O
the	O
mammary	B-Cancer
carcinoma	I-Cancer
experiment	O
,	O
vascular	B-Multi-tissue_structure
permeability	O
measured	O
by	O
means	O
of	O
B22956	B-Simple_chemical
/	I-Simple_chemical
1	I-Simple_chemical
in	O
the	O
animals	O
treated	O
with	O
the	O
antiangiogenic	O
drug	O
(	O
0	O
.	O
0043317	O
+	O
/	O
-	O
0	O
.	O
0040418	O
ml	O
/	O
min	O
(	O
-	O
1	O
)	O
/	O
ml	O
(	O
-	O
1	O
)	O
)	O
was	O
significantly	O
less	O
than	O
in	O
untreated	O
animals	O
(	O
0	O
.	O
0090460	O
+	O
/	O
-	O
0	O
.	O
0043680	O
ml	O
/	O
min	O
(	O
-	O
1	O
)	O
/	O
ml	O
(	O
-	O
1	O
)	O
)	O
,	O
whereas	O
no	O
significant	O
difference	O
was	O
observed	O
with	O
Gd	B-Simple_chemical
-	I-Simple_chemical
DTPA	I-Simple_chemical
-	I-Simple_chemical
albumin	I-Simple_chemical
(	O
13	O
.	O
14	O
+	O
/	O
-	O
13	O
.	O
94	O
ml	O
/	O
min	O
(	O
-	O
1	O
)	O
/	O
ml	O
(	O
-	O
1	O
)	O
in	O
treated	O
animals	O
and	O
18	O
.	O
07	O
+	O
/	O
-	O
11	O
.	O
92	O
ml	O
/	O
min	O
(	O
-	O
1	O
)	O
/	O
ml	O
(	O
-	O
1	O
)	O
in	O
untreated	O
animals	O
)	O
.	O

In	O
the	O
colon	B-Cancer
carcinoma	I-Cancer
experiment	O
,	O
mean	O
permeability	O
and	O
perfusion	O
decreased	O
by	O
51	O
%	O
(	O
from	O
5	O
.	O
2	O
+	O
/	O
-	O
1	O
.	O
1	O
to	O
2	O
.	O
5	O
+	O
/	O
-	O
0	O
.	O
8	O
ml	O
/	O
100	O
ml	O
)	O
and	O
59	O
%	O
(	O
from	O
0	O
.	O
00165	O
+	O
/	O
-	O
5	O
.	O
1	O
to	O
0	O
.	O
0067	O
+	O
/	O
-	O
4	O
.	O
8	O
ml	O
/	O
min	O
(	O
-	O
1	O
)	O
/	O
ml	O
(	O
-	O
1	O
)	O
of	O
tissue	B-Tissue
)	O
,	O
respectively	O
,	O
in	O
all	O
animals	O
after	O
antiangiogenic	O
drug	O
administration	O
.	O

CONCLUSIONS	O
:	O
DCE	O
-	O
MRI	O
permits	O
a	O
noninvasive	O
evaluation	O
of	O
tumour	B-Cancer
microcirculation	O
and	O
in	O
particular	O
of	O
its	O
dynamic	O
characteristics	O
and	O
vascularity	O
before	O
and	O
after	O
antiangiogenic	O
treatment	O
.	O

Angiogenesis	O
in	O
the	O
caprine	B-Organism
caruncles	B-Organism_subdivision
in	O
non	O
-	O
pregnant	O
and	O
pregnant	O
normal	O
and	O
swainsonine	B-Simple_chemical
-	O
treated	O
does	B-Organism
.	O

Microvascular	B-Tissue
corrosion	O
casts	O
of	O
caruncles	B-Organism_subdivision
from	O
non	O
-	O
pregnant	O
and	O
pregnant	O
doe	O
goats	B-Organism
at	O
4	O
,	O
7	O
,	O
10	O
,	O
13	O
,	O
16	O
,	O
and	O
18	O
weeks	O
were	O
examined	O
with	O
scanning	O
electron	O
microscopy	O
.	O

The	O
internal	O
convex	O
surface	O
of	O
the	O
caruncles	B-Organism_subdivision
of	O
non	O
-	O
pregnant	O
does	B-Organism
was	O
covered	O
with	O
capillary	B-Multi-tissue_structure
meshes	I-Multi-tissue_structure
of	O
regular	O
diameter	O
and	O
form	O
,	O
without	O
crypts	O
.	O

As	O
pregnancy	O
advanced	O
the	O
complexity	O
of	O
the	O
vasculature	B-Multi-tissue_structure
increased	O
:	O
at	O
4	O
weeks	O
the	O
surface	O
showed	O
a	O
pattern	O
of	O
ridges	O
separated	O
by	O
troughs	O
.	O

At	O
later	O
stages	O
,	O
branches	B-Multi-tissue_structure
of	O
radial	B-Multi-tissue_structure
arteries	I-Multi-tissue_structure
penetrated	O
the	O
periphery	O
forming	O
an	O
extensive	O
mesh	O
of	O
capillaries	B-Tissue
on	O
the	O
concave	O
surface	O
.	O

Capillary	B-Tissue
diameters	O
increased	O
significantly	O
during	O
pregnancy	O
,	O
especially	O
after	O
4	O
weeks	O
,	O
when	O
large	O
flattened	O
sinusoids	B-Tissue
formed	O
.	O

These	O
sinusoids	B-Tissue
had	O
a	O
great	O
deal	O
of	O
surface	O
area	O
for	O
potential	O
contact	O
with	O
the	O
fetal	B-Developing_anatomical_structure
component	O
.	O

The	O
caprine	B-Organism
placenta	B-Organ
is	O
usually	O
considered	O
to	O
have	O
increased	O
interhemal	B-Immaterial_anatomical_entity
distance	O
compared	O
with	O
endotheliochorial	B-Organ
and	O
hemochorial	B-Organ
types	I-Organ
:	O
our	O
results	O
suggest	O
that	O
the	O
very	O
extensive	O
development	O
of	O
sinusoids	B-Tissue
and	O
crypts	B-Multi-tissue_structure
may	O
compensate	O
for	O
any	O
negative	O
consequences	O
of	O
the	O
placental	B-Organ
architecture	O
.	O

Placental	B-Organ
angiogenesis	O
,	O
which	O
is	O
physiologically	O
normal	O
,	O
may	O
serve	O
as	O
a	O
general	O
model	O
of	O
this	O
process	O
in	O
other	O
circumstances	O
,	O
such	O
as	O
tumor	B-Cancer
.	O

The	O
effect	O
of	O
swainsonine	B-Simple_chemical
(	O
active	O
compound	O
of	O
locoweed	B-Organism
and	O
a	O
potential	O
anticancer	B-Cancer
drug	O
)	O
on	O
vascular	B-Multi-tissue_structure
development	O
showed	O
no	O
differences	O
in	O
sinusoidal	B-Tissue
diameters	O
at	O
7	O
weeks	O
,	O
but	O
a	O
decrease	O
in	O
capillary	B-Tissue
density	O
was	O
noted	O
.	O

Swainsonine	B-Simple_chemical
caused	O
a	O
great	O
distortion	O
to	O
the	O
vasculature	B-Multi-tissue_structure
at	O
18	O
weeks	O
.	O

The	O
effects	O
of	O
this	O
compound	O
on	O
the	O
vascular	B-Multi-tissue_structure
development	O
lend	O
credibility	O
to	O
its	O
potential	O
as	O
an	O
anticancer	B-Cancer
agent	O
.	O

High	O
-	O
grade	O
clear	B-Cancer
cell	I-Cancer
renal	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
has	O
a	O
higher	O
angiogenic	O
activity	O
than	O
low	O
-	O
grade	O
renal	B-Cancer
cell	I-Cancer
carcinoma	I-Cancer
based	O
on	O
histomorphological	O
quantification	O
and	O
qRT	O
-	O
PCR	O
mRNA	O
expression	O
profile	O
.	O

Clear	B-Cancer
cell	I-Cancer
renal	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
(	O
CC	B-Cancer
-	I-Cancer
RCC	I-Cancer
)	O
is	O
a	O
highly	O
vascularised	O
tumour	B-Cancer
and	O
is	O
therefore	O
an	O
attractive	O
disease	O
to	O
study	O
angiogenesis	O
and	O
to	O
test	O
novel	O
angiogenesis	O
inhibitors	O
in	O
early	O
clinical	O
development	O
.	O

Endothelial	B-Cell
cell	I-Cell
proliferation	O
plays	O
a	O
pivotal	O
role	O
in	O
the	O
process	O
of	O
angiogenesis	O
.	O

The	O
aim	O
of	O
this	O
study	O
was	O
to	O
compare	O
angiogenesis	O
parameters	O
in	O
low	O
nuclear	B-Cellular_component
grade	O
(	O
n	O
=	O
87	O
)	O
vs	O
high	O
nuclear	B-Cellular_component
grade	O
CC	B-Cancer
-	I-Cancer
RCC	I-Cancer
(	O
n	O
=	O
63	O
)	O
.	O

A	O
panel	O
of	O
antibodies	O
was	O
used	O
for	O
immunohistochemistry	O
:	O
CD34	B-Gene_or_gene_product
/	O
Ki	B-Gene_or_gene_product
-	I-Gene_or_gene_product
67	I-Gene_or_gene_product
,	O
carbonic	B-Gene_or_gene_product
anhydrase	I-Gene_or_gene_product
IX	I-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
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
(	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
)	O
and	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
.	O

Vessel	B-Multi-tissue_structure
density	O
(	O
MVD	O
-	O
microvessel	B-Tissue
density	O
)	O
,	O
endothelial	B-Cell
cell	I-Cell
proliferation	O
fraction	O
(	O
ECP	O
%	O
)	O
and	O
tumour	B-Cell
cell	I-Cell
proliferation	O
fraction	O
(	O
TCP	O
%	O
)	O
were	O
assessed	O
.	O

mRNA	O
expression	O
levels	O
of	O
angiogenesis	O
stimulators	O
and	O
inhibitors	O
were	O
determined	O
by	O
quantitative	O
RT	O
-	O
PCR	O
.	O

High	O
-	O
grade	O
CC	B-Cancer
-	I-Cancer
RCC	I-Cancer
showed	O
a	O
higher	O
ECP	O
%	O
(	O
P	O
=	O
0	O
.	O
049	O
)	O
,	O
a	O
higher	O
TCP	O
%	O
(	O
P	O
=	O
0	O
.	O
009	O
)	O
,	O
a	O
higher	O
VEGF	B-Gene_or_gene_product
protein	O
expression	O
(	O
P	O
less	O
than	O
0	O
.	O
001	O
)	O
,	O
a	O
lower	O
MVD	O
(	O
P	O
less	O
than	O
0	O
.	O
001	O
)	O
and	O
a	O
lower	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
protein	O
expression	O
(	O
P	O
=	O
0	O
.	O
002	O
)	O
than	O
low	O
-	O
grade	O
CC	B-Cancer
-	I-Cancer
RCC	I-Cancer
.	O

Growth	O
factor	O
mRNA	O
expression	O
analyses	O
revealed	O
a	O
higher	O
expression	O
of	O
angiopoietin	B-Gene_or_gene_product
2	I-Gene_or_gene_product
in	O
low	O
-	O
grade	O
CC	B-Cancer
-	I-Cancer
RCC	I-Cancer
.	O

Microvessel	B-Tissue
density	O
and	O
ECP	O
%	O
were	O
inversely	O
correlated	O
(	O
Rho	O
=	O
-	O
0	O
.	O
26	O
,	O
P	O
=	O
0	O
.	O
001	O
)	O
.	O

Because	O
of	O
the	O
imperfect	O
association	O
of	O
nuclear	B-Cellular_component
grade	O
and	O
ECP	O
%	O
or	O
MVD	O
,	O
CC	B-Cancer
-	I-Cancer
RCC	I-Cancer
was	O
also	O
grouped	O
based	O
on	O
low	O
/	O
high	O
MVD	O
and	O
ECP	O
%	O
.	O

This	O
analysis	O
revealed	O
a	O
higher	O
expression	O
of	O
vessel	B-Multi-tissue_structure
maturation	O
and	O
stabilisation	O
factors	O
(	O
placental	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
,	O
PDGFB1	B-Gene_or_gene_product
,	O
angiopoietin	B-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
in	O
CC	B-Cancer
-	I-Cancer
RCC	I-Cancer
with	O
high	O
MVD	O
,	O
a	O
group	O
of	O
CC	B-Cancer
-	I-Cancer
RCC	I-Cancer
highly	O
enriched	O
in	O
low	O
nuclear	B-Cellular_component
grade	O
CC	B-Cancer
-	I-Cancer
RCC	I-Cancer
,	O
with	O
low	O
ECP	O
%	O
.	O

Our	O
results	O
suggest	O
heterogeneity	O
in	O
angiogenic	O
activity	O
and	O
vessel	B-Multi-tissue_structure
maturation	O
of	O
CC	B-Cancer
-	I-Cancer
RCC	I-Cancer
,	O
to	O
a	O
large	O
extent	O
linked	O
to	O
nuclear	B-Cellular_component
grade	O
,	O
and	O
,	O
with	O
probable	O
therapeutic	O
implications	O
.	O

Deletion	O
of	O
Tip30	B-Gene_or_gene_product
leads	O
to	O
rapid	O
immortalization	O
of	O
murine	B-Organism
mammary	B-Cell
epithelial	I-Cell
cells	I-Cell
and	O
ductal	B-Pathological_formation
hyperplasia	I-Pathological_formation
in	O
the	O
mammary	B-Organ
gland	I-Organ
.	O

Transformation	O
of	O
mammary	B-Cell
epithelial	I-Cell
cells	I-Cell
(	O
MECs	B-Cell
)	O
from	O
the	O
normal	O
to	O
the	O
neoplastic	O
stage	O
requires	O
the	O
dysregulation	O
of	O
tumor	B-Cancer
suppressor	O
genes	O
and	O
proto	O
-	O
oncogenes	O
.	O

Tip30	B-Gene_or_gene_product
is	O
a	O
tumor	B-Cancer
suppressor	O
that	O
can	O
inhibit	O
estrogen	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
-	O
mediated	O
transcription	O
in	O
MECs	B-Cell
,	O
but	O
its	O
role	O
in	O
MEC	B-Cell
proliferation	O
remains	O
unknown	O
.	O

Here	O
,	O
we	O
show	O
that	O
deleting	O
the	O
Tip30	B-Gene_or_gene_product
gene	O
leads	O
to	O
ductal	B-Pathological_formation
hyperplasia	I-Pathological_formation
in	O
mouse	B-Organism
mammary	B-Organ
glands	I-Organ
early	O
in	O
life	O
and	O
extensive	O
mammary	B-Pathological_formation
hyperplasia	I-Pathological_formation
with	O
age	O
.	O

Tip30	B-Gene_or_gene_product
(	O
-	O
/	O
-	O
)	O
mammary	O
glands	O
transplanted	O
into	O
wild	O
-	O
type	O
mammary	B-Tissue
fat	I-Tissue
pads	I-Tissue
also	O
display	O
mammary	B-Multi-tissue_structure
trees	I-Multi-tissue_structure
with	O
extensive	O
ductal	B-Pathological_formation
hyperplasia	I-Pathological_formation
.	O

Strikingly	O
,	O
Tip30	B-Gene_or_gene_product
deletion	O
promotes	O
proliferation	O
of	O
primary	O
MECs	B-Cell
and	O
results	O
in	O
rapid	O
immortalization	O
of	O
MECs	B-Cell
in	O
vitro	O
relative	O
to	O
wild	B-Cell
-	I-Cell
type	I-Cell
cells	I-Cell
.	O

Gene	O
array	O
analysis	O
identified	O
significant	O
increases	O
in	O
the	O
expression	O
of	O
mammary	B-Tissue
epithelial	I-Tissue
growth	O
factors	O
Wisp2	B-Gene_or_gene_product
and	O
Igf	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
in	O
Tip30	B-Gene_or_gene_product
(	O
-	O
/	O
-	O
)	O
cells	O
.	O

Knockdown	O
of	O
either	O
Wisp2	B-Gene_or_gene_product
or	O
Igf	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
using	O
short	O
interfering	O
RNA	O
dramatically	O
inhibited	O
proliferation	O
of	O
Tip30	B-Gene_or_gene_product
(	O
-	O
/	O
-	O
)	O
cells	O
.	O

Together	O
,	O
these	O
results	O
suggest	O
that	O
Tip30	B-Gene_or_gene_product
is	O
an	O
intrinsic	O
and	O
negative	O
regulator	O
of	O
MEC	B-Cell
proliferation	O
partly	O
through	O
the	O
inhibition	O
of	O
Wisp2	B-Gene_or_gene_product
and	O
Igf	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
expression	O
,	O
and	O
its	O
absence	O
in	O
the	O
mammary	B-Organ
gland	I-Organ
may	O
predispose	O
MECs	B-Cell
to	O
neoplastic	O
transformation	O
.	O

A	O
pivotal	O
role	O
for	O
p53	B-Gene_or_gene_product
:	O
balancing	O
aerobic	O
respiration	O
and	O
glycolysis	O
.	O

The	O
genetic	O
basis	O
of	O
increased	O
glycolytic	O
activity	O
observed	O
in	O
cancer	B-Cell
cells	I-Cell
is	O
likely	O
to	O
be	O
the	O
result	O
of	O
complex	O
interactions	O
of	O
multiple	O
regulatory	O
pathways	O
.	O

Here	O
we	O
review	O
the	O
recent	O
evidence	O
of	O
a	O
simple	O
genetic	O
mechanism	O
by	O
which	O
tumor	B-Cancer
suppressor	O
p53	B-Gene_or_gene_product
regulates	O
mitochondrial	B-Cellular_component
respiration	O
with	O
secondary	O
changes	O
in	O
glycolysis	O
that	O
are	O
reminiscent	O
of	O
the	O
Warburg	O
effect	O
.	O

The	O
biological	O
significance	O
of	O
this	O
regulation	O
of	O
the	O
two	O
major	O
pathways	O
of	O
energy	O
generation	O
by	O
p53	B-Gene_or_gene_product
remains	O
to	O
be	O
seen	O
.	O

The	O
contribution	O
of	O
Harold	O
F	O
.	O
Dvorak	O
to	O
the	O
study	O
of	O
tumor	B-Cancer
angiogenesis	O
and	O
stroma	B-Tissue
generation	O
mechanisms	O
.	O

In	O
1983	O
,	O
Harold	O
Dvorak	O
and	O
his	O
colleagues	O
were	O
the	O
first	O
to	O
show	O
that	O
tumor	B-Cell
cells	I-Cell
secreted	O
vascular	B-Gene_or_gene_product
permeability	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VPF	B-Gene_or_gene_product
)	O
and	O
that	O
a	O
blocking	O
antibody	O
to	O
VPF	B-Gene_or_gene_product
could	O
prevent	O
the	O
edema	B-Pathological_formation
and	O
fluid	B-Organism_substance
accumulation	O
that	O
is	O
characteristic	O
of	O
human	B-Organism
cancers	B-Cancer
.	O

In	O
1986	O
,	O
Dvorak	O
went	O
on	O
to	O
demonstrate	O
that	O
VPF	B-Gene_or_gene_product
was	O
secreted	O
by	O
a	O
variety	O
of	O
human	B-Organism
tumor	B-Cell
cell	I-Cell
lines	I-Cell
and	O
proposed	O
that	O
VPF	B-Gene_or_gene_product
was	O
in	O
part	O
responsible	O
for	O
the	O
abnormal	O
vasculature	B-Multi-tissue_structure
seen	O
in	O
human	B-Organism
tumors	B-Cancer
.	O

As	O
a	O
result	O
,	O
he	O
and	O
other	O
investigators	O
demonstrated	O
that	O
VPF	B-Gene_or_gene_product
was	O
capable	O
of	O
stimulating	O
endothelial	B-Cell
cell	I-Cell
growth	O
and	O
angiogenesis	O
.	O

These	O
fundamental	O
discoveries	O
led	O
to	O
additional	O
research	O
conducted	O
by	O
Napoleone	O
Ferrara	O
and	O
his	O
laboratory	O
,	O
confirming	O
the	O
cloning	O
of	O
VPF	B-Gene_or_gene_product
and	O
renaming	O
the	O
protein	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

In	O
1986	O
,	O
Dvorak	O
proposed	O
that	O
by	O
secreting	O
VPF	B-Gene_or_gene_product
,	O
tumors	B-Cancer
induce	O
angiogenesis	O
by	O
turning	O
on	O
the	O
wound	B-Pathological_formation
healing	O
response	O
.	O

He	O
noted	O
that	O
wounds	B-Pathological_formation
,	O
like	O
tumors	B-Cancer
,	O
secrete	O
VPF	B-Gene_or_gene_product
,	O
causing	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
to	O
leak	O
plasma	B-Organism_substance
fibrinogen	B-Gene_or_gene_product
,	O
which	O
stimulates	O
blood	B-Multi-tissue_structure
vessel	I-Multi-tissue_structure
growth	O
and	O
provides	O
a	O
matrix	O
on	O
which	O
they	O
can	O
spread	O
.	O

Unlike	O
wounds	B-Pathological_formation
,	O
however	O
,	O
that	O
turn	O
off	O
VPF	B-Gene_or_gene_product
production	O
after	O
healing	O
,	O
tumors	B-Cancer
did	O
not	O
turn	O
off	O
their	O
VPF	B-Gene_or_gene_product
production	O
and	O
instead	O
continued	O
to	O
make	O
large	O
amounts	O
of	O
VPF	B-Gene_or_gene_product
,	O
allowing	O
malignant	B-Cell
cells	I-Cell
to	O
continue	O
to	O
induce	O
new	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
and	O
so	O
to	O
grow	O
and	O
spread	O
.	O

Thus	O
,	O
tumors	B-Cancer
behave	O
like	O
wounds	B-Pathological_formation
that	O
fail	O
to	O
heal	O
.	O

This	O
work	O
is	O
again	O
extremely	O
significant	O
for	O
patients	B-Organism
worldwide	O
,	O
as	O
Dvorak	O
'	O
s	O
scientific	O
research	O
is	O
leading	O
his	O
colleagues	O
all	O
over	O
the	O
world	O
to	O
examine	O
how	O
to	O
treat	O
a	O
tumor	B-Cancer
through	O
its	O
blood	B-Organism_substance
supply	O
.	O

Melissoidesin	B-Simple_chemical
G	I-Simple_chemical
,	O
a	O
diterpenoid	O
purified	O
from	O
Isodon	B-Organism
melissoides	I-Organism
,	O
induces	O
leukemic	B-Cell
-	I-Cell
cell	I-Cell
apoptosis	O
through	O
induction	O
of	O
redox	O
imbalance	O
and	O
exhibits	O
synergy	O
with	O
other	O
anticancer	B-Cancer
agents	O
.	O

Melissoidesin	B-Simple_chemical
G	I-Simple_chemical
(	O
MOG	B-Simple_chemical
)	O
is	O
a	O
new	O
diterpenoid	O
purified	O
from	O
Isodon	B-Organism
melissoides	I-Organism
,	O
a	O
plant	O
used	O
in	O
Chinese	O
traditional	O
medicine	O
as	O
antitumor	B-Cancer
and	O
anti	O
-	O
inflammatory	O
agents	O
.	O

In	O
our	O
study	O
,	O
MOG	B-Simple_chemical
was	O
shown	O
to	O
specifically	O
inhibit	O
the	O
growth	O
of	O
human	B-Organism
leukemia	B-Cell
cell	I-Cell
lines	I-Cell
and	O
primary	O
acute	B-Cell
myeloid	I-Cell
leukemia	I-Cell
(	I-Cell
AML	I-Cell
)	I-Cell
blasts	I-Cell
via	O
induction	O
of	O
apoptosis	O
,	O
with	O
the	O
evidence	O
of	O
mitochondrial	B-Cellular_component
DeltaPsim	O
loss	O
,	O
reactive	B-Simple_chemical
oxygen	I-Simple_chemical
species	I-Simple_chemical
production	O
,	O
caspases	B-Gene_or_gene_product
activation	O
and	O
nuclear	B-Cellular_component
fragmentation	O
.	O

Furthermore	O
,	O
it	O
was	O
shown	O
that	O
thiol	B-Simple_chemical
-	O
containing	O
antioxidants	O
completely	O
blocked	O
MOG	B-Simple_chemical
-	O
induced	O
mitochondrial	B-Cellular_component
DeltaPsim	O
loss	O
and	O
subsequent	O
cell	B-Cell
apoptosis	O
,	O
while	O
the	O
inhibition	O
of	O
apoptosis	O
by	O
benzyloxy	B-Simple_chemical
-	I-Simple_chemical
carbonyl	I-Simple_chemical
-	I-Simple_chemical
Val	I-Simple_chemical
-	I-Simple_chemical
Ala	I-Simple_chemical
-	I-Simple_chemical
Asp	I-Simple_chemical
-	I-Simple_chemical
fluoromethylketone	I-Simple_chemical
only	O
partially	O
attenuated	O
mitochondrial	B-Cellular_component
DeltaPsim	O
loss	O
,	O
indicating	O
that	O
MOG	B-Simple_chemical
-	O
induced	O
redox	O
imbalance	O
is	O
an	O
early	O
event	O
upstream	O
to	O
mitochondrial	B-Cellular_component
DeltaPsim	O
loss	O
and	O
caspase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
activation	O
.	O

Consistently	O
,	O
it	O
was	O
found	O
that	O
MOG	B-Simple_chemical
rapidly	O
decreased	O
the	O
intracellular	B-Immaterial_anatomical_entity
glutathione	B-Simple_chemical
(	O
GSH	B-Simple_chemical
)	O
content	O
in	O
a	O
dose	O
-	O
dependent	O
manner	O
and	O
the	O
significance	O
of	O
GSH	B-Simple_chemical
depletion	O
in	O
MOG	B-Simple_chemical
-	O
induced	O
apoptosis	O
was	O
further	O
supported	O
by	O
the	O
protective	O
effects	O
of	O
tert	B-Simple_chemical
-	I-Simple_chemical
butylhydroquinone	I-Simple_chemical
(	O
tBHQ	B-Simple_chemical
)	O
and	O
the	O
facilitative	O
effects	O
of	O
DL	B-Simple_chemical
-	I-Simple_chemical
buthionine	I-Simple_chemical
(	I-Simple_chemical
S	I-Simple_chemical
,	I-Simple_chemical
R	I-Simple_chemical
)	I-Simple_chemical
-	I-Simple_chemical
sulfoximine	I-Simple_chemical
(	O
BSO	B-Simple_chemical
)	O
.	O

Furthermore	O
,	O
it	O
was	O
showed	O
that	O
GSH	B-Simple_chemical
depletion	O
induced	O
by	O
MOG	B-Simple_chemical
rendered	O
some	B-Cell
leukemia	I-Cell
cell	I-Cell
lines	I-Cell
more	O
sensitive	O
to	O
arsenic	B-Simple_chemical
trioxide	I-Simple_chemical
(	O
As2O3	B-Simple_chemical
)	O
,	O
doxorubicin	B-Simple_chemical
or	O
cisplatin	B-Simple_chemical
.	O

Additionally	O
,	O
the	O
synergistic	O
apoptotic	O
effects	O
of	O
MOG	B-Simple_chemical
with	O
As2O3	B-Simple_chemical
were	O
detected	O
in	O
HL	B-Cell
-	I-Cell
60	I-Cell
and	O
primary	B-Cell
AML	I-Cell
cells	I-Cell
,	O
but	O
not	O
in	O
normal	O
cells	B-Cell
,	O
suggesting	O
the	O
selective	O
toxicity	O
of	O
their	O
combination	O
to	O
the	O
malignant	B-Cell
cells	I-Cell
.	O

Together	O
,	O
we	O
proposed	O
that	O
MOG	B-Simple_chemical
alone	O
or	O
administered	O
with	O
other	O
anticancer	B-Cancer
agents	O
may	O
provide	O
a	O
novel	O
therapeutic	O
strategy	O
for	O
leukemia	B-Cancer
.	O

Hedgehog	B-Gene_or_gene_product
signaling	O
in	O
the	O
murine	B-Organism
melanoma	B-Cancer
microenvironment	O
.	O

The	O
Hedgehog	B-Gene_or_gene_product
intercellular	O
signaling	O
pathway	O
regulates	O
cell	B-Cell
proliferation	O
and	O
differentiation	O
.	O

This	O
pathway	O
has	O
been	O
implicated	O
to	O
play	O
a	O
role	O
in	O
the	O
pathogenesis	O
of	O
cancer	B-Cancer
and	O
in	O
embryonic	B-Multi-tissue_structure
blood	I-Multi-tissue_structure
vessel	I-Multi-tissue_structure
development	O
.	O

In	O
the	O
current	O
study	O
,	O
Hedgehog	B-Gene_or_gene_product
signaling	O
in	O
tumor	B-Cancer
related	O
vasculature	B-Multi-tissue_structure
and	O
microenvironment	O
was	O
examined	O
using	O
human	B-Cell
umbilical	I-Cell
vein	I-Cell
endothelial	I-Cell
cells	I-Cell
and	O
B16F0	B-Cell
(	O
murine	B-Organism
melanoma	B-Cancer
)	O
tumors	B-Cancer
models	O
.	O

Use	O
of	O
exogenous	O
Sonic	B-Gene_or_gene_product
hedgehog	I-Gene_or_gene_product
(	O
Shh	B-Gene_or_gene_product
)	O
peptide	O
significantly	O
increased	O
BrdU	B-Simple_chemical
incorporation	O
in	O
endothelial	B-Cell
cells	I-Cell
in	O
vitro	O
by	O
a	O
factor	O
of	O
2	O
(	O
P	O
less	O
than	O
0	O
.	O
001	O
)	O
.	O

The	O
Hedgehog	B-Gene_or_gene_product
pathway	O
antagonist	O
cyclopamine	B-Simple_chemical
effectively	O
reduced	O
Shh	B-Gene_or_gene_product
-	O
induced	O
proliferation	O
to	O
control	O
levels	O
.	O

To	O
study	O
Hedgehog	B-Gene_or_gene_product
signaling	O
in	O
vivo	O
a	O
hind	B-Cancer
limb	I-Cancer
tumor	I-Cancer
model	O
with	O
the	O
B16F0	B-Cell
cell	I-Cell
line	I-Cell
was	O
used	O
.	O

Treatment	O
with	O
25	O
mg	O
/	O
kg	O
cyclopamine	B-Simple_chemical
significantly	O
attenuated	O
BrdU	B-Simple_chemical
incorporation	O
in	O
tumor	B-Cell
cells	I-Cell
threefold	O
(	O
P	O
less	O
than	O
0	O
.	O
001	O
)	O
,	O
in	O
tumor	B-Cancer
related	O
endothelial	B-Cell
cells	I-Cell
threefold	O
(	O
P	O
=	O
0	O
.	O
004	O
)	O
,	O
and	O
delayed	O
tumor	B-Cancer
growth	O
by	O
4	O
days	O
.	O

Immunohistochemistry	O
revealed	O
that	O
the	O
Hedgehog	B-Gene_or_gene_product
receptor	O
Patched	B-Gene_or_gene_product
was	O
localized	O
to	O
the	O
tumor	B-Tissue
stroma	I-Tissue
and	O
that	O
B16F0	B-Cell
cells	I-Cell
expressed	O
Shh	B-Gene_or_gene_product
peptide	O
.	O

Furthermore	O
,	O
mouse	B-Organism
embryonic	B-Cell
fibroblasts	I-Cell
required	O
the	O
presence	O
of	O
B16F0	B-Cell
cells	I-Cell
to	O
express	O
Patched	B-Gene_or_gene_product
in	O
a	O
co	O
-	O
culture	O
assay	O
system	O
.	O

These	O
studies	O
indicate	O
that	O
Shh	B-Gene_or_gene_product
peptide	O
produced	O
by	O
melanoma	B-Cell
cells	I-Cell
induces	O
Patched	B-Gene_or_gene_product
expression	O
in	O
fibroblasts	B-Cell
.	O

To	O
study	O
tumor	B-Cancer
related	O
angiogenesis	O
a	O
vascular	B-Multi-tissue_structure
window	O
model	O
was	O
used	O
to	O
monitor	O
tumor	B-Cancer
vascularity	O
.	O

Treatment	O
with	O
cyclopamine	B-Simple_chemical
significantly	O
attenuated	O
vascular	B-Multi-tissue_structure
formation	O
by	O
a	O
factor	O
of	O
2	O
.	O
5	O
(	O
P	O
less	O
than	O
0	O
.	O
001	O
)	O
and	O
altered	O
vascular	B-Multi-tissue_structure
morphology	O
.	O

Furthermore	O
,	O
cyclopamine	B-Simple_chemical
reduced	O
tumor	B-Multi-tissue_structure
blood	I-Multi-tissue_structure
vessel	I-Multi-tissue_structure
permeability	O
to	O
FITC	B-Simple_chemical
labeled	O
dextran	B-Simple_chemical
while	O
having	O
no	O
effect	O
on	O
normal	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
.	O

These	O
studies	O
suggest	O
that	O
Hedgehog	B-Gene_or_gene_product
signaling	O
regulates	O
melanoma	B-Cancer
related	O
vascular	B-Multi-tissue_structure
formation	O
and	O
function	O
.	O

Expression	O
of	O
epidermal	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
,	O
transforming	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
and	O
Ki	B-Gene_or_gene_product
-	I-Gene_or_gene_product
67	I-Gene_or_gene_product
in	O
relationship	O
to	O
malignant	O
transformation	O
of	O
pleomorphic	B-Pathological_formation
adenoma	I-Pathological_formation
.	O

CONCLUSION	O
:	O
Quantitative	O
assessment	O
is	O
more	O
sensitive	O
as	O
a	O
measure	O
of	O
cellular	B-Cell
protein	O
content	O
as	O
compared	O
with	O
standard	O
optical	O
density	O
measurements	O
.	O

The	O
data	O
support	O
the	O
hypothesis	O
that	O
increased	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
and	O
transforming	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	I-Gene_or_gene_product
TGF	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
expression	O
is	O
associated	O
with	O
early	O
events	O
in	O
malignant	O
transformation	O
of	O
pleomorphic	B-Pathological_formation
adenoma	I-Pathological_formation
(	O
PA	B-Pathological_formation
)	O
.	O

OBJECTIVE	O
:	O
In	O
the	O
present	O
study	O
,	O
we	O
attempted	O
to	O
identify	O
EGFR	B-Gene_or_gene_product
and	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
expression	O
and	O
Ki	B-Gene_or_gene_product
-	I-Gene_or_gene_product
67	I-Gene_or_gene_product
index	O
in	O
carcinoma	B-Cancer
ex	I-Cancer
-	I-Cancer
pleomorphic	I-Cancer
adenoma	I-Cancer
(	O
Ca	B-Cancer
ex	I-Cancer
-	I-Cancer
PA	I-Cancer
)	O
and	O
PA	B-Pathological_formation
.	O

We	O
also	O
compared	O
the	O
presence	O
of	O
EGFR	B-Gene_or_gene_product
and	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
and	O
Ki	B-Gene_or_gene_product
-	I-Gene_or_gene_product
67	I-Gene_or_gene_product
index	O
with	O
clinical	O
data	O
.	O

MATERIALS	O
AND	O
METHODS	O
:	O
The	O
tissues	B-Tissue
were	O
stained	O
with	O
monoclonal	O
antibodies	O
to	O
EGFR	B-Gene_or_gene_product
,	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
and	O
Ki	B-Gene_or_gene_product
-	I-Gene_or_gene_product
67	I-Gene_or_gene_product
.	O

The	O
results	O
were	O
analysed	O
using	O
quantitative	O
immunohistochemical	O
analysis	O
.	O

We	O
also	O
analysed	O
the	O
association	O
of	O
patients	B-Organism
'	O
prognosis	O
with	O
clinical	O
parameters	O
and	O
the	O
histological	O
classification	O
of	O
the	O
carcinomatous	B-Cancer
component	I-Cancer
.	O

RESULTS	O
:	O
As	O
regards	O
the	O
association	O
of	O
patients	B-Organism
'	O
prognosis	O
with	O
EGFR	B-Gene_or_gene_product
staining	O
and	O
Ki	B-Gene_or_gene_product
-	I-Gene_or_gene_product
67	I-Gene_or_gene_product
index	O
,	O
a	O
significant	O
increase	O
was	O
observed	O
in	O
patients	B-Organism
who	O
died	O
or	O
had	O
residual	O
disease	O
compared	O
with	O
patients	B-Organism
who	O
were	O
alive	O
without	O
disease	O
.	O

In	O
the	O
immunohistochemical	O
analysis	O
of	O
EGFR	B-Gene_or_gene_product
and	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
and	O
Ki67	B-Gene_or_gene_product
index	O
,	O
a	O
significant	O
increase	O
was	O
observed	O
in	O
Ca	B-Cancer
ex	I-Cancer
-	I-Cancer
PA	I-Cancer
,	O
especially	O
with	O
adenocarcinoma	B-Cancer
,	O
compared	O
with	O
PA	B-Pathological_formation
and	O
sialadenitis	O
.	O

CX3CR1	B-Gene_or_gene_product
-	O
dependent	O
subretinal	B-Cell
microglia	I-Cell
cell	I-Cell
accumulation	O
is	O
associated	O
with	O
cardinal	O
features	O
of	O
age	O
-	O
related	O
macular	B-Tissue
degeneration	O
.	O

The	O
role	O
of	O
retinal	B-Cell
microglial	I-Cell
cells	I-Cell
(	O
MCs	B-Cell
)	O
in	O
age	O
-	O
related	O
macular	B-Tissue
degeneration	O
(	O
AMD	O
)	O
is	O
unclear	O
.	O

Here	O
we	O
demonstrated	O
that	O
all	O
retinal	B-Cell
MCs	I-Cell
express	O
CX3C	B-Gene_or_gene_product
chemokine	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
CX3CR1	B-Gene_or_gene_product
)	O
and	O
that	O
homozygosity	O
for	O
the	O
CX3CR1	B-Gene_or_gene_product
M280	O
allele	O
,	O
which	O
is	O
associated	O
with	O
impaired	O
cell	B-Cell
migration	O
,	O
increases	O
the	O
risk	O
of	O
AMD	O
.	O

In	O
humans	B-Organism
with	O
AMD	O
,	O
MCs	B-Cell
accumulated	O
in	O
the	O
subretinal	B-Immaterial_anatomical_entity
space	I-Immaterial_anatomical_entity
at	O
sites	O
of	O
retinal	B-Multi-tissue_structure
degeneration	O
and	O
choroidal	B-Multi-tissue_structure
neovascularization	O
(	O
CNV	O
)	O
.	O

In	O
CX3CR1	B-Gene_or_gene_product
-	O
deficient	O
mice	B-Organism
,	O
MCs	B-Cell
accumulated	O
subretinally	B-Immaterial_anatomical_entity
with	O
age	O
and	O
albino	O
background	O
and	O
after	O
laser	O
impact	O
preceding	O
retinal	B-Multi-tissue_structure
degeneration	O
.	O

Raising	O
the	O
albino	O
mice	B-Organism
in	O
the	O
dark	O
prevented	O
both	O
events	O
.	O

The	O
appearance	O
of	O
lipid	B-Simple_chemical
-	O
bloated	O
subretinal	B-Cell
MCs	I-Cell
was	O
drusen	B-Pathological_formation
-	O
like	O
on	O
funduscopy	O
of	O
senescent	O
mice	B-Organism
,	O
and	O
CX3CR1	B-Gene_or_gene_product
-	O
dependent	O
MC	B-Cell
accumulation	O
was	O
associated	O
with	O
an	O
exacerbation	O
of	O
experimental	O
CNV	O
.	O

These	O
results	O
show	O
that	O
CX3CR1	B-Gene_or_gene_product
-	O
dependent	O
accumulation	O
of	O
subretinal	B-Cell
MCs	I-Cell
evokes	O
cardinal	O
features	O
of	O
AMD	O
.	O

These	O
findings	O
reveal	O
what	O
we	O
believe	O
to	O
be	O
a	O
novel	O
pathogenic	O
process	O
with	O
important	O
implications	O
for	O
the	O
development	O
of	O
new	O
therapies	O
for	O
AMD	O
.	O

Conditional	O
deletion	O
of	O
Smad1	B-Gene_or_gene_product
and	O
Smad5	B-Gene_or_gene_product
in	O
somatic	B-Cell
cells	I-Cell
of	O
male	O
and	O
female	O
gonads	B-Organ
leads	O
to	O
metastatic	O
tumor	B-Cancer
development	O
in	O
mice	B-Organism
.	O

The	O
transforming	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
(	O
TGFbeta	B-Gene_or_gene_product
)	O
family	O
has	O
critical	O
roles	O
in	O
the	O
regulation	O
of	O
fertility	O
.	O

In	O
addition	O
,	O
the	O
pathogenesis	O
of	O
some	O
human	B-Organism
cancers	B-Cancer
is	O
attributed	O
to	O
misregulation	O
of	O
TGFbeta	B-Gene_or_gene_product
function	O
and	O
SMAD2	B-Gene_or_gene_product
or	O
SMAD4	B-Gene_or_gene_product
mutations	O
.	O

There	O
are	O
limited	O
mouse	B-Organism
models	O
for	O
the	O
BMP	B-Gene_or_gene_product
signaling	I-Gene_or_gene_product
SMADs	I-Gene_or_gene_product
(	I-Gene_or_gene_product
BR	I-Gene_or_gene_product
-	I-Gene_or_gene_product
SMADs	I-Gene_or_gene_product
)	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
5	B-Gene_or_gene_product
,	O
and	O
8	B-Gene_or_gene_product
because	O
of	O
embryonic	B-Developing_anatomical_structure
lethality	O
and	O
suspected	O
genetic	O
redundancy	O
.	O

Using	O
tissue	B-Tissue
-	O
specific	O
ablation	O
in	O
mice	B-Organism
,	O
we	O
deleted	O
the	O
BR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
SMADs	I-Gene_or_gene_product
from	O
somatic	B-Cell
cells	I-Cell
of	O
ovaries	B-Organ
and	O
testes	B-Organ
.	O

Single	O
conditional	O
knockouts	O
for	O
Smad1	B-Gene_or_gene_product
or	O
Smad5	B-Gene_or_gene_product
or	O
mice	B-Organism
homozygous	O
null	O
for	O
Smad8	B-Gene_or_gene_product
are	O
viable	O
and	O
fertile	O
.	O

Female	O
double	O
Smad1	B-Gene_or_gene_product
Smad5	B-Gene_or_gene_product
and	O
triple	O
Smad1	B-Gene_or_gene_product
Smad5	B-Gene_or_gene_product
Smad8	B-Gene_or_gene_product
conditional	O
knockout	O
mice	O
become	O
infertile	O
and	O
develop	O
metastatic	O
granulosa	B-Cancer
cell	I-Cancer
tumors	I-Cancer
.	O

Male	O
double	O
Smad1	B-Gene_or_gene_product
Smad5	B-Gene_or_gene_product
conditional	O
knockout	O
mice	O
are	O
fertile	O
but	O
demonstrate	O
metastatic	O
testicular	B-Cancer
tumor	I-Cancer
development	O
.	O

Microarray	O
analysis	O
indicated	O
significant	O
alterations	O
in	O
expression	O
of	O
genes	O
related	O
to	O
the	O
TGFbeta	B-Gene_or_gene_product
pathway	O
,	O
as	O
well	O
as	O
genes	O
involved	O
in	O
infertility	O
and	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
production	O
.	O

These	O
data	O
strongly	O
implicate	O
the	O
BR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
SMADs	I-Gene_or_gene_product
as	O
part	O
of	O
a	O
critical	O
developmental	O
pathway	O
in	O
ovaries	B-Organ
and	O
testis	B-Organ
that	O
,	O
when	O
disrupted	O
,	O
leads	O
to	O
malignant	O
transformation	O
.	O

[	O
Clinical	O
significance	O
of	O
interleukin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
6	I-Gene_or_gene_product
(	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
6	I-Gene_or_gene_product
)	O
as	O
a	O
prognostic	O
factor	O
of	O
cancer	B-Cancer
disease	O
]	O

Interleukin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
6	I-Gene_or_gene_product
(	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
6	I-Gene_or_gene_product
)	O
is	O
proinflammatory	O
cytokine	O
that	O
produces	O
multifunctional	O
effects	O
.	O

It	O
is	O
also	O
involved	O
in	O
the	O
regulation	O
of	O
immune	O
reactions	O
,	O
hematopoiesis	O
and	O
inflammatory	O
state	O
.	O

Interleukin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
6	I-Gene_or_gene_product
has	O
been	O
shown	O
to	O
be	O
associated	O
with	O
tumor	B-Cancer
progression	O
including	O
inhibition	O
of	O
cancer	B-Cell
cells	I-Cell
apoptosis	O
and	O
stimulation	O
of	O
angiogenesis	O
.	O

Anti	B-Gene_or_gene_product
-	I-Gene_or_gene_product
IL	I-Gene_or_gene_product
-	I-Gene_or_gene_product
6	I-Gene_or_gene_product
therapy	O
is	O
a	O
new	O
strategy	O
in	O
the	O
inflammatory	O
autoimmune	O
diseases	O
and	O
cancer	B-Cancer
.	O

Clinical	O
studies	O
have	O
shown	O
elevated	O
serum	B-Organism_substance
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
6	I-Gene_or_gene_product
concentrations	O
in	O
patients	B-Organism
with	O
endometrial	B-Cancer
cancer	I-Cancer
,	O
non	B-Cancer
-	I-Cancer
small	I-Cancer
cell	I-Cancer
lung	I-Cancer
carcinoma	I-Cancer
,	O
colorectal	B-Cancer
cancer	I-Cancer
,	O
renal	B-Cancer
cell	I-Cancer
carcinoma	I-Cancer
,	O
breast	B-Cancer
and	O
ovarian	B-Cancer
cancer	I-Cancer
.	O

Serum	B-Organism_substance
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
6	I-Gene_or_gene_product
levels	O
correlate	O
with	O
tumor	B-Cancer
stage	O
,	O
and	O
survival	O
of	O
patients	B-Organism
.	O

In	O
this	O
article	O
we	O
have	O
focused	O
on	O
a	O
role	O
of	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
6	I-Gene_or_gene_product
as	O
a	O
prognostic	O
factor	O
in	O
several	O
malignancies	B-Cancer
such	O
as	O
colorectal	B-Cancer
cancer	I-Cancer
,	O
breast	B-Cancer
cancer	I-Cancer
,	O
gastric	B-Cancer
cancer	I-Cancer
and	O
pancreatic	B-Cancer
cancer	I-Cancer
.	O

Expression	O
of	O
CDK4	B-Gene_or_gene_product
or	O
CDK2	B-Gene_or_gene_product
in	O
mouse	B-Organism
oral	B-Immaterial_anatomical_entity
cavity	I-Immaterial_anatomical_entity
is	O
retained	O
in	O
adult	O
pituitary	B-Organ
with	O
distinct	O
effects	O
on	O
tumorigenesis	O
.	O

The	O
keratin	B-Gene_or_gene_product
5	I-Gene_or_gene_product
(	O
K5	B-Gene_or_gene_product
)	O
promoter	O
drives	O
transgenic	O
expression	O
to	O
the	O
basal	B-Cell
cell	I-Cell
layer	O
of	O
stratified	O
epithelia	B-Tissue
.	O

Surprisingly	O
,	O
analysis	O
of	O
K5CDK4	B-Gene_or_gene_product
and	O
K5CDK2	B-Gene_or_gene_product
transgenic	O
mouse	O
embryos	B-Developing_anatomical_structure
showed	O
CDK4	B-Gene_or_gene_product
and	O
CDK2	B-Gene_or_gene_product
expression	O
not	O
only	O
in	O
the	O
expected	O
tissues	B-Tissue
,	O
but	O
also	O
in	O
the	O
adenohypophysis	B-Organ
.	O

This	O
organ	B-Organ
is	O
derived	O
from	O
an	O
upwards	O
growth	O
of	O
the	O
primitive	O
oropharynx	B-Anatomical_system
,	O
a	O
K5	B-Gene_or_gene_product
-	O
expressing	O
tissue	B-Tissue
.	O

We	O
show	O
that	O
transgenic	O
expression	O
of	O
CDKs	B-Gene_or_gene_product
in	O
the	O
embryonic	B-Developing_anatomical_structure
oral	I-Developing_anatomical_structure
ectoderm	I-Developing_anatomical_structure
is	O
specifically	O
retained	O
in	O
undifferentiated	O
cells	B-Cell
from	O
the	O
pars	B-Multi-tissue_structure
intermedia	I-Multi-tissue_structure
of	O
the	O
adenohypophysis	B-Organ
.	O

Interestingly	O
,	O
we	O
found	O
that	O
K5CDK4	B-Gene_or_gene_product
mice	O
show	O
a	O
decreased	O
number	O
of	O
pituitary	B-Cell
stem	I-Cell
cells	I-Cell
,	O
even	O
though	O
CDK4	B-Gene_or_gene_product
is	O
not	O
expressed	O
in	O
the	O
stem	B-Cell
cells	I-Cell
but	O
in	O
transit	B-Cell
-	I-Cell
amplifying	I-Cell
(	I-Cell
TA	I-Cell
)	I-Cell
-	I-Cell
like	I-Cell
cells	I-Cell
.	O

Interestingly	O
,	O
CDK4	B-Gene_or_gene_product
-	O
expressing	O
cells	B-Cell
,	O
but	O
not	O
CDK2	B-Gene_or_gene_product
-	O
expressing	O
cells	B-Cell
,	O
strongly	O
synergize	O
with	O
lack	O
of	O
p27	B-Gene_or_gene_product
(	O
Kip1	B-Gene_or_gene_product
)	O
to	O
generate	O
pituitary	B-Cancer
carcinomas	I-Cancer
that	O
appear	O
with	O
shortened	O
latency	O
and	O
are	O
drastically	O
more	O
aggressive	O
than	O
those	O
arising	O
in	O
p27	B-Gene_or_gene_product
(	O
-	O
/	O
-	O
)	O
mice	O
.	O

Thus	O
,	O
we	O
show	O
that	O
deregulation	O
of	O
CDK	B-Gene_or_gene_product
expression	O
in	O
the	O
primitive	B-Tissue
oral	I-Tissue
epithelium	I-Tissue
plays	O
a	O
unique	O
function	O
,	O
providing	O
a	O
selective	O
advantage	O
that	O
gives	O
rise	O
to	O
transgene	B-Cell
-	I-Cell
positive	I-Cell
TA	I-Cell
-	I-Cell
like	I-Cell
pituitary	I-Cell
cells	I-Cell
.	O

Furthermore	O
,	O
retention	O
of	O
CDK4	B-Gene_or_gene_product
in	O
these	O
TA	B-Cell
-	I-Cell
like	I-Cell
pituitary	I-Cell
cells	I-Cell
synergizes	O
with	O
loss	O
of	O
p27	B-Gene_or_gene_product
(	O
Kip1	B-Gene_or_gene_product
)	O
to	O
induce	O
pituitary	B-Cancer
adenocarcinomas	I-Cancer
.	O

This	O
model	O
suggests	O
that	O
forced	O
expression	O
of	O
CDK4	B-Gene_or_gene_product
sensitizes	O
cells	B-Cell
and	O
synergizes	O
with	O
a	O
second	O
change	O
resulting	O
in	O
tumor	B-Cancer
development	O
.	O

Survivin	B-Gene_or_gene_product
expression	O
in	O
breast	B-Cancer
carcinoma	I-Cancer
:	O
correlation	O
with	O
apoptosis	O
and	O
prognosis	O
.	O

BACKGROUND	O
:	O
Survivin	B-Gene_or_gene_product
is	O
a	O
novel	O
inhibitor	O
of	O
apoptosis	O
commonly	O
detected	O
in	O
tissues	B-Tissue
during	O
fetal	B-Developing_anatomical_structure
development	O
and	O
in	O
cancer	B-Cancer
,	O
but	O
not	O
usually	O
in	O
normal	B-Tissue
tissues	I-Tissue
.	O

It	O
has	O
been	O
suggested	O
that	O
the	O
expression	O
of	O
this	O
protein	O
may	O
be	O
of	O
prognostic	O
significance	O
in	O
gastric	B-Cancer
,	O
colorectal	B-Cancer
,	O
and	O
bladder	B-Cancer
carcinomas	I-Cancer
.	O

We	O
assessed	O
survivin	B-Gene_or_gene_product
expression	O
in	O
breast	B-Cancer
carcinomas	I-Cancer
correlating	O
results	O
with	O
expression	O
of	O
other	O
antiapoptotic	O
(	O
bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
x	I-Gene_or_gene_product
)	O
and	O
proapoptotic	O
(	O
bax	B-Gene_or_gene_product
)	O
markers	O
,	O
with	O
prognostic	O
parameters	O
,	O
and	O
with	O
prognosis	O
.	O

DESIGN	O
:	O
Paraffin	B-Simple_chemical
-	O
embedded	O
sections	B-Cancer
of	O
37	O
breast	B-Cancer
carcinomas	I-Cancer
were	O
immunostained	O
for	O
survivin	B-Gene_or_gene_product
,	O
bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
x	I-Gene_or_gene_product
,	O
and	O
bax	B-Gene_or_gene_product
.	O

Expression	O
was	O
evaluated	O
in	O
normal	O
breast	B-Tissue
tissue	I-Tissue
and	O
carcinoma	B-Cancer
,	O
nuclei	B-Cellular_component
and	O
cytoplasm	B-Organism_substance
,	O
as	O
intensity	O
(	O
0	O
to	O
3	O
+	O
)	O
,	O
and	O
percentage	O
of	O
positive	O
cells	B-Cell
.	O

RESULTS	O
:	O
Survivin	B-Gene_or_gene_product
expression	O
was	O
noted	O
in	O
30	O
(	O
81	O
%	O
)	O
of	O
breast	B-Cancer
carcinomas	I-Cancer
,	O
and	O
in	O
normal	O
breast	B-Tissue
tissue	I-Tissue
,	O
in	O
nuclei	B-Cellular_component
,	O
and	O
cytoplasm	B-Organism_substance
.	O

There	O
was	O
a	O
significant	O
correlation	O
(	O
P	O
=	O
0	O
.	O
022	O
)	O
between	O
survivin	B-Gene_or_gene_product
and	O
bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
x	I-Gene_or_gene_product
expression	O
;	O
survivin	B-Gene_or_gene_product
and	O
bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
x	I-Gene_or_gene_product
tended	O
to	O
correlate	O
with	O
overall	O
survival	O
(	O
P	O
=	O
0	O
.	O
072	O
and	O
0	O
.	O
075	O
,	O
respectively	O
)	O
,	O
but	O
not	O
with	O
disease	O
-	O
free	O
survival	O
(	O
P	O
=	O
0	O
.	O
19	O
and	O
0	O
.	O
18	O
,	O
respectively	O
)	O
.	O

There	O
was	O
no	O
correlation	O
of	O
survivin	B-Gene_or_gene_product
with	O
bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
or	O
bax	B-Gene_or_gene_product
expression	O
,	O
or	O
with	O
other	O
prognostic	O
parameters	O
(	O
age	O
,	O
tumor	B-Cancer
size	O
,	O
histologic	O
type	O
,	O
histologic	O
grade	O
,	O
nodal	B-Multi-tissue_structure
status	O
,	O
and	O
tumor	B-Cancer
stage	O
)	O
(	O
P	O
>	O
0	O
.	O
05	O
)	O
.	O

CONCLUSION	O
:	O
The	O
majority	O
(	O
81	O
%	O
)	O
of	O
breast	B-Cancer
carcinomas	I-Cancer
show	O
survivin	B-Gene_or_gene_product
expression	O
which	O
correlates	O
with	O
bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
x	I-Gene_or_gene_product
,	O
another	O
antiapoptotic	O
marker	O
,	O
and	O
both	O
markers	O
tend	O
to	O
correlate	O
with	O
prognosis	O
.	O

Functional	O
significance	O
of	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
a	I-Gene_or_gene_product
in	O
human	B-Organism
ovarian	B-Cancer
carcinoma	I-Cancer
:	O
role	O
in	O
vasculogenic	O
mimicry	O
.	O

Ovarian	B-Cancer
cancer	I-Cancer
is	O
a	O
silent	O
killer	O
,	O
and	O
shows	O
early	O
extensive	O
tumor	B-Cancer
invasion	O
and	O
peritoneal	B-Multi-tissue_structure
metastasis	O
.	O

The	O
microcirculation	O
of	O
most	O
tumors	B-Cancer
includes	O
cooperation	O
of	O
pre	O
-	O
existing	O
vessels	B-Multi-tissue_structure
,	O
intussusceptive	O
microvascular	B-Tissue
growth	O
,	O
postnatal	O
vasculogenesis	O
,	O
glomeruloid	B-Multi-tissue_structure
angiogenesis	O
and	O
vasculogenic	O
mimicry	O
(	O
VM	O
)	O
.	O

VM	O
is	O
critical	O
for	O
a	O
tumor	B-Cancer
blood	B-Organism_substance
supply	O
and	O
is	O
asscociated	O
with	O
aggressive	O
features	O
and	O
metastasis	O
.	O

Our	O
studies	O
highlight	O
the	O
plasticity	O
of	O
aggressive	O
human	B-Organism
ovarian	B-Cell
carcinoma	I-Cell
cells	I-Cell
and	O
call	O
into	O
question	O
the	O
underlying	O
significance	O
of	O
their	O
ability	O
to	O
form	O
VM	O
in	O
vitro	O
induced	O
by	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
a	I-Gene_or_gene_product
.	O

These	O
studies	O
also	O
show	O
their	O
clinicalpathological	O
features	O
of	O
the	O
cancers	B-Cancer
with	O
human	B-Organism
Paraffin	B-Simple_chemical
-	O
embedded	O
tumor	B-Tissue
tissue	I-Tissue
samples	I-Tissue
.	O

Results	O
show	O
that	O
the	O
process	O
:	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
a	I-Gene_or_gene_product
-	O
-	O
greater	O
than	O
EphA2	B-Gene_or_gene_product
-	O
-	O
greater	O
than	O
MMPs	B-Gene_or_gene_product
-	O
-	O
greater	O
than	O
VM	O
is	O
the	O
main	O
pathway	O
for	O
VM	O
formation	O
and	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
a	I-Gene_or_gene_product
appears	O
to	O
play	O
an	O
important	O
role	O
in	O
the	O
formation	O
of	O
VM	O
based	O
on	O
our	O
in	O
vitro	O
assays	O
and	O
clinical	O
immunohistochemical	O
analyses	O
.	O

VM	O
-	O
targeting	O
strategies	O
for	O
ovarian	B-Cancer
cancer	I-Cancer
include	O
anti	B-Gene_or_gene_product
-	I-Gene_or_gene_product
VEGF	I-Gene_or_gene_product
-	I-Gene_or_gene_product
a	I-Gene_or_gene_product
treatment	O
,	O
knocking	O
down	O
the	O
EphA2	B-Gene_or_gene_product
gene	O
and	O
using	O
antibodies	B-Gene_or_gene_product
against	I-Gene_or_gene_product
human	I-Gene_or_gene_product
MMPs	I-Gene_or_gene_product
if	O
the	O
tumor	B-Cancer
is	O
VM	O
positive	O
.	O

This	O
strategy	O
may	O
be	O
of	O
significant	O
value	O
in	O
laying	O
the	O
foundation	O
for	O
a	O
more	O
explicit	O
anti	O
-	O
tumor	B-Cancer
angiogenesis	O
therapy	O
.	O

A	O
novel	O
role	O
of	O
thrombospondin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
in	O
cervical	B-Multi-tissue_structure
carcinogenesis	O
:	O
inhibit	O
stroma	B-Tissue
reaction	O
by	O
inhibiting	O
activated	O
fibroblasts	B-Cell
from	O
invading	O
cancer	B-Cancer
.	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
a	O
potent	O
angiogenesis	O
inhibitor	O
,	O
has	O
been	O
shown	O
to	O
exert	O
different	O
biological	O
functions	O
on	O
various	O
cell	B-Cell
types	O
.	O

Here	O
,	O
we	O
investigate	O
the	O
role	O
of	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
in	O
tumor	B-Cancer
-	O
stroma	B-Tissue
reaction	O
,	O
which	O
is	O
mainly	O
characterized	O
by	O
fibroblast	B-Cell
activation	O
to	O
create	O
a	O
permissive	O
microenvironment	O
for	O
tumor	B-Cancer
progression	O
.	O

Immunohistochemistry	O
examinations	O
in	O
the	O
human	B-Organism
surgical	B-Cancer
specimens	I-Cancer
have	O
shown	O
that	O
a	O
downregulation	O
of	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
during	O
the	O
progression	O
of	O
cervical	B-Multi-tissue_structure
carcinogenesis	O
was	O
accompanied	O
by	O
an	O
emergence	O
in	O
the	O
upregulation	O
of	O
stroma	B-Gene_or_gene_product
markers	I-Gene_or_gene_product
,	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
smooth	I-Gene_or_gene_product
muscle	I-Gene_or_gene_product
actin	I-Gene_or_gene_product
(	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
SMA	I-Gene_or_gene_product
)	O
and	O
desmin	B-Gene_or_gene_product
.	O

Transfection	O
of	O
SiHa	B-Cell
cervical	I-Cell
cancer	I-Cell
cells	I-Cell
with	O
a	O
plasmid	O
expressing	O
the	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
protein	O
exhibited	O
antiangiogenic	O
activity	O
in	O
vitro	O
and	O
resulted	O
in	O
reduced	O
tumor	B-Cancer
growth	O
in	O
severe	O
combined	O
immunodeficiency	O
(	O
SCID	O
)	O
mice	B-Organism
,	O
which	O
was	O
accompanied	O
by	O
a	O
decrease	O
in	O
tumor	B-Cancer
vascularization	O
and	O
lower	O
expressions	O
of	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
SMA	I-Gene_or_gene_product
and	O
desmin	B-Gene_or_gene_product
than	O
those	O
in	O
the	O
vector	O
controls	O
.	O

Transfection	O
with	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
purified	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
added	O
to	O
NIH3T3	B-Cell
cells	I-Cell
did	O
not	O
alter	O
the	O
protein	O
levels	O
of	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
SMA	I-Gene_or_gene_product
and	O
desmin	B-Gene_or_gene_product
but	O
significantly	O
inhibited	O
matrix	B-Gene_or_gene_product
metalloprotease	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
activity	O
.	O

Transforming	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
(	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
)	O
,	O
a	O
major	O
factor	O
in	O
the	O
activation	O
of	O
fibroblasts	B-Cell
,	O
increased	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
SMA	I-Gene_or_gene_product
and	O
desmin	B-Gene_or_gene_product
expression	O
and	O
the	O
ability	O
of	O
cell	B-Cell
migration	O
and	O
invasion	O
in	O
NIH3T3	B-Cell
cells	I-Cell
.	O

The	O
increased	O
migration	O
ability	O
and	O
the	O
invasive	O
ability	O
into	O
tumor	B-Cancer
cluster	O
of	O
TGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
-	O
treated	O
NIH3T3	B-Cell
cells	I-Cell
were	O
dose	O
dependently	O
inhibited	O
by	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

In	O
contrast	O
,	O
ectopic	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
expression	O
in	O
SiHa	B-Cell
cells	I-Cell
has	O
little	O
effect	O
on	O
the	O
invasive	O
ability	O
of	O
the	O
NIH3T3	B-Cell
cells	I-Cell
.	O

Together	O
,	O
our	O
findings	O
demonstrate	O
a	O
novel	O
role	O
of	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
to	O
inhibit	O
tumor	B-Cancer
-	O
stroma	B-Tissue
reaction	O
that	O
could	O
be	O
attributed	O
to	O
the	O
blockage	O
of	O
activated	O
fibroblasts	B-Cell
from	O
invading	O
cancer	B-Cell
cells	I-Cell
.	O

Targeting	O
a	O
tumor	B-Cancer
-	O
specific	O
laminin	B-Gene_or_gene_product
domain	O
critical	O
for	O
human	B-Organism
carcinogenesis	O
.	O

Laminin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
332	I-Gene_or_gene_product
is	O
critical	O
for	O
squamous	B-Cancer
cell	I-Cancer
carcinoma	I-Cancer
(	O
SCC	B-Cancer
)	O
tumorigenesis	O
,	O
but	O
targeting	O
it	O
for	O
cancer	B-Cancer
therapy	O
has	O
been	O
unachievable	O
due	O
to	O
key	O
role	O
of	O
laminin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
332	I-Gene_or_gene_product
in	O
promoting	O
tissue	B-Tissue
integrity	O
.	O

Here	O
,	O
we	O
show	O
that	O
a	O
portion	O
of	O
laminin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
332	I-Gene_or_gene_product
,	O
termed	O
G45	B-Gene_or_gene_product
,	O
which	O
is	O
proteolytically	O
removed	O
and	O
absent	O
in	O
normal	B-Tissue
tissues	I-Tissue
,	O
is	O
prominently	O
expressed	O
in	O
most	O
human	B-Organism
SCC	B-Cancer
tumors	I-Cancer
and	O
plays	O
an	O
important	O
role	O
in	O
human	B-Organism
SCC	B-Cancer
tumorigenesis	O
.	O

Primary	O
human	B-Organism
keratinocytes	O
lacking	O
G45	B-Gene_or_gene_product
(	O
DeltaG45	B-Cell
)	O
showed	O
alterations	O
of	O
basal	O
receptor	O
organization	O
,	O
impaired	O
matrix	O
deposition	O
,	O
and	O
increased	O
migration	O
.	O

After	O
SCC	B-Cancer
transformation	O
,	O
the	O
absence	O
of	O
G45	B-Gene_or_gene_product
domain	O
in	O
DeltaG45	B-Gene_or_gene_product
cells	O
was	O
associated	O
with	O
deficient	O
extracellular	B-Gene_or_gene_product
signal	I-Gene_or_gene_product
-	I-Gene_or_gene_product
regulated	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
and	O
phosphotidylinositol	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
pathway	O
activation	O
,	O
impaired	O
invasion	O
,	O
deficient	O
metalloproteinase	B-Gene_or_gene_product
activity	O
,	O
and	O
absent	O
tumorgenicity	O
in	O
vivo	O
.	O

Expression	O
of	O
G45	B-Gene_or_gene_product
or	O
activated	O
PI3K	B-Gene_or_gene_product
subunit	O
in	O
DeltaG45	B-Gene_or_gene_product
cells	O
reversed	O
these	O
abnormalities	O
.	O

G45	B-Gene_or_gene_product
antibody	O
treatment	O
induced	O
SCC	B-Cancer
tumor	I-Cancer
apoptosis	O
,	O
decreased	O
SCC	B-Cancer
tumor	I-Cancer
proliferation	O
,	O
and	O
markedly	O
impaired	O
human	B-Organism
SCC	B-Cancer
tumorigenesis	O
in	O
vivo	O
without	O
affecting	O
normal	B-Tissue
tissue	I-Tissue
adhesion	O
.	O

These	O
results	O
show	O
a	O
remarkable	O
selectivity	O
of	O
expression	O
and	O
function	O
for	O
laminin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
332	I-Gene_or_gene_product
G45	B-Gene_or_gene_product
in	O
human	B-Organism
SCC	B-Cancer
tumorigenesis	O
and	O
implicate	O
it	O
as	O
a	O
specific	O
target	O
for	O
anticancer	B-Cancer
therapy	O
.	O

Management	O
of	O
early	O
and	O
advanced	O
colorectal	B-Cancer
cancer	I-Cancer
:	O
therapeutic	O
issues	O
.	O

PURPOSE	O
:	O
The	O
staging	O
of	O
colorectal	B-Cancer
cancer	I-Cancer
,	O
therapeutic	O
decision	O
making	O
in	O
the	O
management	O
of	O
early	O
and	O
advanced	O
colorectal	B-Cancer
cancer	I-Cancer
,	O
and	O
dilemmas	O
posed	O
by	O
drug	O
-	O
related	O
toxicity	O
are	O
discussed	O
.	O

SUMMARY	O
:	O
Staging	O
of	O
colorectal	B-Cancer
cancer	I-Cancer
occurs	O
after	O
surgery	O
and	O
is	O
based	O
on	O
the	O
extent	O
of	O
disease	O
invasiveness	O
and	O
dissemination	O
.	O

Surgery	O
is	O
the	O
primary	O
treatment	O
for	O
stage	O
I	O
disease	O
.	O

Adjuvant	O
chemotherapy	O
is	O
recommended	O
after	O
resection	O
in	O
selected	O
high	O
-	O
risk	O
patients	B-Organism
with	O
stage	O
II	O
disease	O
and	O
in	O
all	O
patients	B-Organism
with	O
stage	O
III	O
disease	O
.	O

Convenience	O
of	O
administration	O
,	O
tolerability	O
,	O
and	O
patient	B-Organism
factors	O
not	O
necessarily	O
age	O
may	O
be	O
considerations	O
in	O
decisions	O
about	O
adjuvant	O
therapy	O
after	O
resection	O
.	O

Treatment	O
of	O
stage	O
IV	O
colorectal	B-Cancer
cancer	I-Cancer
is	O
based	O
on	O
the	O
type	O
of	O
prior	O
therapy	O
and	O
patient	B-Organism
-	O
specific	O
factors	O
.	O

Recently	O
,	O
significant	O
improvements	O
in	O
survival	O
have	O
been	O
achieved	O
through	O
the	O
use	O
of	O
combination	O
chemotherapy	O
and	O
monoclonal	O
antibody	O
regimens	O
.	O

Bevacizumab	B-Simple_chemical
in	O
combination	O
with	O
chemotherapy	O
is	O
first	O
-	O
line	O
therapy	O
for	O
stage	O
IV	O
disease	O
.	O

Age	O
alone	O
should	O
not	O
preclude	O
the	O
use	O
of	O
chemotherapy	O
in	O
stage	O
IV	O
colorectal	B-Cancer
cancer	I-Cancer
,	O
although	O
the	O
ability	O
to	O
tolerate	O
drug	O
-	O
related	O
toxicity	O
may	O
be	O
a	O
consideration	O
.	O

The	O
optimal	O
duration	O
of	O
chemotherapy	O
in	O
patients	B-Organism
with	O
early	O
and	O
metastatic	O
colorectal	B-Cancer
cancer	I-Cancer
is	O
unclear	O
.	O

CONCLUSION	O
:	O
The	O
optimal	O
approach	O
to	O
the	O
treatment	O
of	O
colorectal	B-Cancer
cancer	I-Cancer
depends	O
on	O
several	O
considerations	O
,	O
including	O
patient	B-Organism
-	O
specific	O
factors	O
.	O

Serum	B-Organism_substance
and	O
urine	B-Organism_substance
levels	O
of	O
interleukin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
in	O
patients	B-Organism
with	O
non	B-Cancer
-	I-Cancer
Hodgkin	I-Cancer
'	I-Cancer
s	I-Cancer
lymphoma	I-Cancer
.	O

Angiogenesis	O
plays	O
an	O
important	O
role	O
in	O
many	O
types	O
of	O
cancer	B-Cancer
.	O

Interleukin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
(	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
)	O
is	O
known	O
to	O
be	O
a	O
pro	O
-	O
inflammatory	O
and	O
pro	O
-	O
angiogenic	O
cytokine	O
,	O
and	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
has	O
been	O
reported	O
to	O
be	O
associated	O
with	O
tumor	B-Cancer
progression	O
,	O
prognosis	O
and	O
survival	O
in	O
several	O
types	O
of	O
cancers	B-Cancer
.	O

However	O
,	O
the	O
role	O
of	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
in	O
non	B-Cancer
-	I-Cancer
Hodgkin	I-Cancer
'	I-Cancer
s	I-Cancer
lymphoma	I-Cancer
(	O
NHL	B-Cancer
)	O
has	O
not	O
been	O
fully	O
determined	O
.	O

Here	O
,	O
we	O
evaluated	O
the	O
usefulness	O
of	O
measuring	O
serum	B-Organism_substance
and	O
urine	B-Organism_substance
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
levels	O
in	O
patients	B-Organism
with	O
NHL	B-Cancer
.	O

We	O
developed	O
reference	O
intervals	O
for	O
serum	B-Organism_substance
and	O
urine	B-Organism_substance
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
level	O
in	O
131	O
control	O
individuals	O
.	O

We	O
measured	O
serum	B-Organism_substance
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
and	O
urine	B-Organism_substance
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
levels	O
in	O
patients	B-Organism
with	O
NHL	B-Cancer
,	O
and	O
we	O
compared	O
the	O
concentrations	O
with	O
those	O
of	O
control	O
individuals	O
.	O

The	O
reference	O
intervals	O
for	O
serum	B-Organism_substance
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
and	O
urine	B-Organism_substance
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
corrected	O
by	O
creatinine	B-Simple_chemical
(	O
Cr	B-Simple_chemical
)	O
were	O
15	O
.	O
9	O
-	O
430	O
.	O
3	O
pg	O
/	O
mL	O
and	O
0	O
.	O
0	O
-	O
28	O
.	O
4	O
pg	O
/	O
mg	O
Cr	B-Simple_chemical
,	O
respectively	O
.	O

The	O
concentrations	O
of	O
urine	B-Organism_substance
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
/	O
Cr	B-Simple_chemical
were	O
significantly	O
higher	O
in	O
patients	B-Organism
than	O
in	O
controls	O
(	O
48	O
.	O
9	O
+	O
/	O
-	O
194	O
.	O
4	O
vs	O
.	O
5	O
.	O
2	O
+	O
/	O
-	O
13	O
.	O
8	O
pg	O
/	O
mg	O
Cr	B-Simple_chemical
,	O
P	O
less	O
than	O
0	O
.	O
001	O
)	O
.	O

However	O
,	O
there	O
were	O
no	O
significant	O
differences	O
in	O
serum	B-Organism_substance
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
concentrations	O
between	O
NHL	B-Cancer
patients	B-Organism
and	O
controls	O
(	O
159	O
.	O
2	O
+	O
/	O
-	O
40	O
.	O
4	O
vs	O
.	O
99	O
.	O
6	O
+	O
/	O
-	O
107	O
.	O
1	O
pg	O
/	O
mL	O
;	O
P	O
=	O
0	O
.	O
099	O
)	O
.	O

Receiver	O
operating	O
characteristic	O
(	O
ROC	O
)	O
analysis	O
gave	O
0	O
.	O
83	O
and	O
0	O
.	O
43	O
ROC	O
area	O
values	O
for	O
urine	B-Organism_substance
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
/	O
Cr	B-Simple_chemical
and	O
serum	B-Organism_substance
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
,	O
respectively	O
.	O

There	O
was	O
no	O
correlation	O
between	O
the	O
serum	B-Organism_substance
and	O
urine	B-Organism_substance
concentrations	O
of	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
and	O
clinical	O
variables	O
,	O
the	O
only	O
exception	O
being	O
the	O
international	O
prognostic	O
index	O
(	O
IPI	O
)	O
,	O
which	O
showed	O
a	O
marginal	O
correlation	O
with	O
urine	B-Organism_substance
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
/	O
Cr	B-Simple_chemical
levels	O
(	O
P	O
=	O
0	O
.	O
07	O
)	O
.	O

This	O
study	O
indicated	O
that	O
urine	B-Organism_substance
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
/	O
Cr	B-Simple_chemical
levels	O
might	O
be	O
useful	O
as	O
a	O
diagnostic	O
marker	O
of	O
NHL	B-Cancer
.	O

Microscopic	O
technique	O
for	O
the	O
detection	O
of	O
nitric	B-Simple_chemical
oxide	I-Simple_chemical
-	O
dependent	O
angiogenesis	O
in	O
an	O
animal	O
model	O
.	O

Nitric	B-Simple_chemical
oxide	I-Simple_chemical
(	O
NO	B-Simple_chemical
)	O
plays	O
an	O
important	O
role	O
in	O
maintaining	O
vascular	B-Multi-tissue_structure
homeostasis	O
.	O

The	O
importance	O
of	O
NO	B-Simple_chemical
in	O
the	O
vasculature	B-Anatomical_system
is	O
demonstrated	O
by	O
several	O
experimental	O
conditions	O
,	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
-	O
induced	O
angiogenesis	O
.	O

Thus	O
,	O
the	O
NO	B-Simple_chemical
metabolic	O
pathway	O
in	O
endothelial	B-Cell
cells	I-Cell
could	O
be	O
one	O
of	O
the	O
main	O
contributing	O
factors	O
for	O
angiogenesis	O
.	O

Although	O
several	O
methods	O
have	O
been	O
used	O
for	O
measuring	O
in	O
vitro	O
angiogenesis	O
,	O
a	O
proper	O
technique	O
has	O
not	O
been	O
developed	O
for	O
identifying	O
in	O
vivo	O
NO	B-Simple_chemical
-	O
dependent	O
angiogenesis	O
.	O

This	O
chapter	O
provides	O
a	O
new	O
intravital	O
microscopic	O
method	O
for	O
detecting	O
and	O
measuring	O
NO	B-Simple_chemical
-	O
dependent	O
angiogenesis	O
in	O
a	O
mouse	B-Organism
model	O
.	O

This	O
technique	O
showed	O
strong	O
abdominal	B-Organism_subdivision
neovascularization	O
in	O
wild	O
-	O
type	O
mice	B-Organism
,	O
but	O
not	O
eNOS	B-Gene_or_gene_product
knockout	O
mice	O
,	O
locally	O
injected	O
with	O
VEGF	B-Gene_or_gene_product
,	O
as	O
well	O
as	O
stimulation	O
of	O
angiogenesis	O
in	O
NO	B-Simple_chemical
donor	O
-	O
injected	O
mice	B-Organism
.	O

This	O
technique	O
also	O
revealed	O
the	O
inhibitory	O
effect	O
of	O
the	O
NOS	B-Gene_or_gene_product
inhibitor	O
N	B-Simple_chemical
(	I-Simple_chemical
G	I-Simple_chemical
)	I-Simple_chemical
-	I-Simple_chemical
iminoethyl	I-Simple_chemical
-	I-Simple_chemical
L	I-Simple_chemical
-	I-Simple_chemical
ornithine	I-Simple_chemical
in	O
VEGF	B-Gene_or_gene_product
-	O
mediated	O
in	O
vivo	O
angiogenesis	O
.	O

This	O
chapter	O
describes	O
intravital	O
microscopy	O
as	O
a	O
new	O
imaging	O
technique	O
for	O
detecting	O
NO	B-Simple_chemical
-	O
dependent	O
angiogenesis	O
in	O
an	O
animal	O
model	O
.	O

Effects	O
of	O
cyclooxygenase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
non	O
-	O
selective	O
and	O
selective	O
inhibitors	O
on	O
proliferation	O
inhibition	O
and	O
apoptosis	O
induction	O
of	O
esophageal	B-Cell
squamous	I-Cell
carcinoma	I-Cell
cells	I-Cell
.	O

The	O
objective	O
of	O
this	O
study	O
is	O
to	O
investigate	O
the	O
effects	O
of	O
aspirin	B-Simple_chemical
and	O
nimesulide	B-Simple_chemical
on	O
cell	B-Cell
proliferation	O
,	O
apoptosis	O
and	O
its	O
potential	O
mechanisms	O
in	O
EC9706	B-Cell
and	O
EC109	B-Cell
esophageal	I-Cell
squamous	I-Cell
carcinoma	I-Cell
cells	I-Cell
.	O

EC9706	B-Cell
and	O
EC109	B-Cell
cells	I-Cell
were	O
incubated	O
with	O
varying	O
concentrations	O
of	O
aspirin	B-Simple_chemical
and	O
nimesulide	B-Simple_chemical
,	O
and	O
the	O
effects	O
on	O
cell	B-Cell
proliferation	O
and	O
apoptosis	O
were	O
monitored	O
by	O
3	B-Simple_chemical
-	I-Simple_chemical
(	I-Simple_chemical
4	I-Simple_chemical
,	I-Simple_chemical
5	I-Simple_chemical
-	I-Simple_chemical
dimethyl	I-Simple_chemical
-	I-Simple_chemical
thiazol	I-Simple_chemical
-	I-Simple_chemical
2yl	I-Simple_chemical
)	I-Simple_chemical
-	I-Simple_chemical
2	I-Simple_chemical
,	I-Simple_chemical
5	I-Simple_chemical
-	I-Simple_chemical
diphenyltetrazolium	I-Simple_chemical
bromide	I-Simple_chemical
and	O
flow	O
cytometry	O
.	O

Reverse	O
transcription	O
-	O
polymerase	O
chain	O
reaction	O
and	O
Western	O
blot	O
assays	O
were	O
used	O
to	O
investigate	O
expression	O
of	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
Bax	B-Gene_or_gene_product
.	O

Prostaglandin	B-Gene_or_gene_product
E2	I-Gene_or_gene_product
production	O
was	O
measured	O
by	O
enzyme	O
linked	O
immunosorbent	O
assay	O
.	O

Pretreatment	O
with	O
aspirin	B-Simple_chemical
and	O
nimesulide	B-Simple_chemical
inhibited	O
EC9706	B-Cell
and	O
EC109	B-Cell
cell	I-Cell
growth	O
in	O
a	O
time	O
and	O
dose	O
-	O
dependent	O
manner	O
,	O
accompanied	O
with	O
a	O
decrease	O
of	O
prostaglandin	B-Gene_or_gene_product
E2	I-Gene_or_gene_product
production	O
.	O

In	O
EC9706	B-Cell
cells	I-Cell
,	O
the	O
mechanism	O
of	O
aspirin	B-Simple_chemical
and	O
nimesulide	B-Simple_chemical
induced	O
growth	O
inhibition	O
was	O
a	O
consequence	O
of	O
cell	B-Cell
cycle	O
arrest	O
at	O
the	O
G	O
(	O
0	O
)	O
/	O
G	O
(	O
1	O
)	O
check	O
point	O
.	O

In	O
EC109	B-Cell
cells	I-Cell
,	O
growth	O
arrest	O
was	O
by	O
induction	O
of	O
apoptosis	O
,	O
associated	O
with	O
downregulation	O
of	O
Bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
but	O
not	O
Bax	B-Gene_or_gene_product
.	O

In	O
conclusion	O
,	O
aspirin	B-Simple_chemical
and	O
nimesulide	B-Simple_chemical
could	O
inhibit	O
cell	B-Cell
proliferation	O
and	O
induce	O
apoptosis	O
in	O
esophageal	B-Cell
squamous	I-Cell
carcinoma	I-Cell
cells	I-Cell
.	O

Cyclooxygenase	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
inhibitor	O
may	O
be	O
a	O
promising	O
therapeutic	O
agent	O
for	O
human	B-Organism
esophageal	B-Cancer
squamous	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
.	O

Neurotrophin	B-Gene_or_gene_product
p75	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
(	O
p75NTR	B-Gene_or_gene_product
)	O
promotes	O
endothelial	B-Cell
cell	I-Cell
apoptosis	O
and	O
inhibits	O
angiogenesis	O
:	O
implications	O
for	O
diabetes	O
-	O
induced	O
impaired	O
neovascularization	O
in	O
ischemic	O
limb	B-Organ
muscles	I-Organ
.	O

Diabetes	O
impairs	O
endothelial	B-Cell
function	O
and	O
reparative	O
neovascularization	O
.	O

The	O
p75	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
of	I-Gene_or_gene_product
neurotrophins	I-Gene_or_gene_product
(	O
p75	B-Gene_or_gene_product
(	I-Gene_or_gene_product
NTR	I-Gene_or_gene_product
)	I-Gene_or_gene_product
)	O
,	O
which	O
is	O
scarcely	O
present	O
in	O
healthy	O
endothelial	B-Cell
cells	I-Cell
(	O
ECs	B-Cell
)	O
,	O
becomes	O
strongly	O
expressed	O
by	O
capillary	B-Cell
ECs	I-Cell
after	O
induction	O
of	O
peripheral	B-Tissue
ischemia	O
in	O
type	O
-	O
1	O
diabetic	O
mice	B-Organism
.	O

Here	O
,	O
we	O
show	O
that	O
gene	O
transfer	O
-	O
induced	O
p75	B-Gene_or_gene_product
(	I-Gene_or_gene_product
NTR	I-Gene_or_gene_product
)	I-Gene_or_gene_product
expression	O
impairs	O
the	O
survival	O
,	O
proliferation	O
,	O
migration	O
,	O
and	O
adhesion	O
capacities	O
of	O
cultured	O
ECs	B-Cell
and	O
endothelial	B-Cell
progenitor	I-Cell
cells	I-Cell
(	O
EPCs	B-Cell
)	O
and	O
inhibits	O
angiogenesis	O
in	O
vitro	O
.	O

Moreover	O
,	O
intramuscular	B-Immaterial_anatomical_entity
p75	B-Gene_or_gene_product
(	I-Gene_or_gene_product
NTR	I-Gene_or_gene_product
)	I-Gene_or_gene_product
gene	O
delivery	O
impairs	O
neovascularization	O
and	O
blood	B-Organism_substance
flow	O
recovery	O
in	O
a	O
mouse	B-Organism
model	O
of	O
limb	B-Organism_subdivision
ischemia	O
.	O

These	O
disturbed	O
functions	O
are	O
associated	O
with	O
suppression	O
of	O
signaling	O
mechanisms	O
implicated	O
in	O
EC	B-Cell
survival	O
and	O
angiogenesis	O
.	O

In	O
fact	O
,	O
p75	B-Gene_or_gene_product
(	I-Gene_or_gene_product
NTR	I-Gene_or_gene_product
)	I-Gene_or_gene_product
depresses	O
the	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
A	I-Gene_or_gene_product
/	O
Akt	B-Gene_or_gene_product
/	O
eNOS	B-Gene_or_gene_product
/	O
NO	B-Simple_chemical
pathway	O
and	O
additionally	O
reduces	O
the	O
mRNA	O
levels	O
of	O
ITGB1	B-Gene_or_gene_product
[	O
beta	B-Gene_or_gene_product
(	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	I-Gene_or_gene_product
integrin	I-Gene_or_gene_product
]	O
,	O
BIRC5	B-Gene_or_gene_product
(	O
survivin	B-Gene_or_gene_product
)	O
,	O
PTTG1	B-Gene_or_gene_product
(	O
securin	B-Gene_or_gene_product
)	O
and	O
VEZF1	B-Gene_or_gene_product
.	O

Diabetic	O
mice	B-Organism
,	O
which	O
typically	O
show	O
impaired	O
postischemic	O
muscular	B-Organ
neovascularization	O
and	O
blood	B-Organism_substance
perfusion	O
recovery	O
,	O
have	O
these	O
defects	O
corrected	O
by	O
intramuscular	B-Immaterial_anatomical_entity
gene	O
transfer	O
of	O
a	O
dominant	O
negative	O
mutant	O
form	O
of	O
p75	B-Gene_or_gene_product
(	I-Gene_or_gene_product
NTR	I-Gene_or_gene_product
)	I-Gene_or_gene_product
.	O

Collectively	O
,	O
our	O
data	O
newly	O
demonstrate	O
the	O
antiangiogenic	O
action	O
of	O
p75	B-Gene_or_gene_product
(	I-Gene_or_gene_product
NTR	I-Gene_or_gene_product
)	I-Gene_or_gene_product
and	O
open	O
new	O
avenues	O
for	O
the	O
therapeutic	O
use	O
of	O
p75	B-Gene_or_gene_product
(	I-Gene_or_gene_product
NTR	I-Gene_or_gene_product
)	I-Gene_or_gene_product
inhibition	O
to	O
combat	O
diabetes	O
-	O
induced	O
microvascular	B-Tissue
liabilities	O
.	O

Tumor	B-Cell
endothelial	I-Cell
cell	I-Cell
targeted	O
cyclic	B-Simple_chemical
RGD	I-Simple_chemical
-	I-Simple_chemical
modified	I-Simple_chemical
heparin	I-Simple_chemical
derivative	I-Simple_chemical
:	O
inhibition	O
of	O
angiogenesis	O
and	O
tumor	B-Cancer
growth	O
.	O

PURPOSE	O
:	O
We	O
prepared	O
tumor	B-Tissue
endothelium	I-Tissue
targeted	O
cRGD	B-Simple_chemical
-	I-Simple_chemical
modified	I-Simple_chemical
heparin	I-Simple_chemical
derivative	I-Simple_chemical
(	O
cRGD	B-Simple_chemical
-	I-Simple_chemical
HL	I-Simple_chemical
)	O
by	O
coupling	O
heparin	B-Simple_chemical
-	I-Simple_chemical
lithocholic	I-Simple_chemical
acid	I-Simple_chemical
(	O
HL	B-Simple_chemical
)	O
with	O
cRGDyK	B-Simple_chemical
,	O
and	O
evaluated	O
inhibition	O
effects	O
of	O
cRGD	B-Simple_chemical
-	I-Simple_chemical
HL	I-Simple_chemical
on	O
angiogenesis	O
and	O
tumor	B-Cancer
growth	O
.	O

METHODS	O
:	O
To	O
evaluate	O
antiangiogenic	O
activity	O
of	O
cRGD	B-Simple_chemical
-	I-Simple_chemical
HL	I-Simple_chemical
,	O
we	O
performed	O
tests	O
on	O
endothelial	B-Cell
cell	I-Cell
adhesion	O
and	O
migration	O
to	O
vitronectin	B-Gene_or_gene_product
,	O
tube	B-Tissue
formation	O
,	O
binding	O
affinity	O
to	O
purified	O
alpha	B-Gene_or_gene_product
(	I-Gene_or_gene_product
v	I-Gene_or_gene_product
)	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
(	I-Gene_or_gene_product
3	I-Gene_or_gene_product
)	I-Gene_or_gene_product
integrin	I-Gene_or_gene_product
,	O
and	O
in	O
vivo	O
Matrigel	O
plug	O
assay	O
.	O

The	O
antitumor	B-Cancer
activity	O
of	O
cRGD	B-Simple_chemical
-	I-Simple_chemical
HL	I-Simple_chemical
was	O
also	O
evaluated	O
by	O
monitoring	O
tumor	B-Cancer
growth	O
and	O
microvessel	B-Tissue
formation	O
in	O
squamous	B-Cancer
cell	I-Cancer
carcinoma	I-Cancer
(	I-Cancer
SCC7	I-Cancer
)	I-Cancer
tumor	I-Cancer
.	O

RESULTS	O
:	O
The	O
cRGD	B-Simple_chemical
-	I-Simple_chemical
HL	I-Simple_chemical
significantly	O
inhibited	O
adhesion	O
and	O
migration	O
of	O
endothelial	B-Cell
cells	I-Cell
to	O
vitronectin	B-Gene_or_gene_product
,	O
and	O
tubular	B-Tissue
structures	I-Tissue
of	O
endothelial	B-Cell
cells	I-Cell
.	O

Compared	O
to	O
cRGDyK	B-Simple_chemical
and	O
HL	B-Simple_chemical
,	O
cRGD	B-Simple_chemical
-	I-Simple_chemical
HL	I-Simple_chemical
has	O
high	O
binding	O
affinity	O
to	O
purified	O
alpha	B-Gene_or_gene_product
(	I-Gene_or_gene_product
v	I-Gene_or_gene_product
)	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
(	I-Gene_or_gene_product
3	I-Gene_or_gene_product
)	I-Gene_or_gene_product
integrin	I-Gene_or_gene_product
.	O

The	O
enhanced	O
antiangiogenic	O
effect	O
of	O
cRGD	B-Simple_chemical
-	I-Simple_chemical
HL	I-Simple_chemical
was	O
confirmed	O
in	O
Matrigel	O
assay	O
by	O
showing	O
the	O
significant	O
inhibition	O
of	O
bFGF	B-Gene_or_gene_product
-	O
driven	O
angiogenesis	O
and	O
blood	B-Multi-tissue_structure
vessel	I-Multi-tissue_structure
formation	O
.	O

It	O
was	O
thought	O
that	O
potent	O
antiangiogenic	O
effect	O
of	O
cRGD	B-Simple_chemical
-	I-Simple_chemical
HL	I-Simple_chemical
was	O
probably	O
due	O
to	O
the	O
interference	O
of	O
alpha	B-Gene_or_gene_product
(	I-Gene_or_gene_product
v	I-Gene_or_gene_product
)	I-Gene_or_gene_product
beta	I-Gene_or_gene_product
(	I-Gene_or_gene_product
3	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	O
mediated	O
interaction	O
,	O
resulting	O
in	O
the	O
enhanced	O
antitumoral	B-Cancer
activity	O
against	O
SCC7	B-Cancer
tumor	I-Cancer
.	O

CONCLUSION	O
:	O
These	O
results	O
demonstrated	O
that	O
cRGD	B-Simple_chemical
-	I-Simple_chemical
modified	I-Simple_chemical
heparin	I-Simple_chemical
derivative	I-Simple_chemical
enhanced	O
anti	O
-	O
angiotherapeutic	O
effects	O
against	O
solid	B-Cancer
tumor	I-Cancer
,	O
and	O
therefore	O
,	O
it	O
could	O
be	O
applied	O
to	O
treat	O
various	O
cancers	B-Cancer
and	O
angiogenic	O
diseases	O
as	O
a	O
potent	O
angiogenesis	O
inhibitor	O
.	O

[	O
Inhibitory	O
effect	O
of	O
adenovirus	B-Organism
-	O
mediated	O
endostatin	B-Gene_or_gene_product
gene	O
transfer	O
on	O
transplanted	O
lung	B-Cancer
carcinoma	I-Cancer
in	O
mice	B-Organism
]	O

OBJECTIVE	O
:	O
To	O
investigate	O
the	O
effect	O
of	O
adenovirus	B-Organism
-	O
mediated	O
endostatin	B-Gene_or_gene_product
gene	O
transfer	O
on	O
transplanted	O
lung	B-Cancer
cancer	I-Cancer
in	O
mice	B-Organism
and	O
its	O
mechanism	O
of	O
action	O
.	O

METHODS	O
:	O
Transplant	B-Cancer
tumor	I-Cancer
model	O
was	O
induced	O
by	O
subcutaneous	O
inoculation	O
of	O
2	O
x	O
10	O
(	O
6	O
)	O
Lewis	B-Cell
lung	I-Cell
cancer	I-Cell
(	I-Cell
LLC	I-Cell
)	I-Cell
cells	I-Cell
into	O
the	O
back	B-Organism_subdivision
of	O
C57BL	B-Organism
/	I-Organism
6	I-Organism
mice	I-Organism
.	O

The	O
mice	B-Organism
were	O
treated	O
by	O
intratumoral	B-Cancer
injection	O
of	O
2	O
x	O
10	O
(	O
9	O
)	O
pfu	O
Ad	B-Gene_or_gene_product
-	I-Gene_or_gene_product
mEndostatin	I-Gene_or_gene_product
.	O

The	O
expression	O
of	O
endostatin	B-Gene_or_gene_product
in	O
situ	O
and	O
its	O
maintaining	O
time	O
were	O
detected	O
by	O
immunohistochemistry	O
and	O
Western	O
Blot	O
,	O
respectively	O
.	O

The	O
endostatin	B-Gene_or_gene_product
level	O
in	O
serum	B-Organism_substance
was	O
determined	O
by	O
ELISA	O
.	O

The	O
inhibition	O
of	O
tumor	B-Cancer
growth	O
and	O
changes	O
of	O
survival	O
were	O
recorded	O
and	O
the	O
microvessel	B-Tissue
density	O
(	O
MVD	O
)	O
was	O
determined	O
by	O
histochemical	O
stainingwith	O
CD31	B-Gene_or_gene_product
and	O
CD105	B-Gene_or_gene_product
antibodies	I-Gene_or_gene_product
.	O

The	O
tumor	B-Cancer
apoptosis	O
was	O
observed	O
by	O
electron	O
microscopy	O
.	O

RESULTS	O
:	O
In	O
comparison	O
with	O
controls	O
,	O
intratumoral	B-Cancer
injection	O
of	O
Ad	B-Gene_or_gene_product
-	I-Gene_or_gene_product
mEndostatin	I-Gene_or_gene_product
significantly	O
inhibited	O
the	O
tumor	B-Cancer
growth	O
and	O
metastasis	O
,	O
and	O
prolonged	O
the	O
survival	O
rate	O
of	O
mice	B-Organism
(	O
P	O
less	O
than	O
0	O
.	O
05	O
)	O
.	O

Strong	O
positive	O
expression	O
of	O
mEndostatin	B-Gene_or_gene_product
was	O
seen	O
in	O
the	O
tumor	B-Tissue
tissue	I-Tissue
after	O
injection	O
of	O
Ad	B-Gene_or_gene_product
-	I-Gene_or_gene_product
mEndostatin	I-Gene_or_gene_product
,	O
immunhistochemically	O
ostained	O
by	O
mouse	B-Organism
endostatin	B-Gene_or_gene_product
monoclonal	I-Gene_or_gene_product
antibody	I-Gene_or_gene_product
,	O
while	O
the	O
control	O
groups	O
showed	O
only	O
very	O
low	O
expression	O
or	O
absence	O
.	O

Serum	B-Organism_substance
endostatin	B-Gene_or_gene_product
concentration	O
was	O
1540	O
+	O
/	O
-	O
560	O
ng	O
/	O
ml	O
at	O
the	O
second	O
week	O
of	O
administration	O
,	O
the	O
expression	O
of	O
endostatin	B-Gene_or_gene_product
diminished	O
a	O
month	O
later	O
.	O

The	O
microvessel	B-Tissue
density	O
(	O
MVD	O
)	O
)	O
decreased	O
from	O
42	O
.	O
4	O
+	O
/	O
-	O
4	O
.	O
8	O
to	O
10	O
.	O
5	O
+	O
/	O
-	O
3	O
.	O
2	O
per	O
x	O
200	O
magnificetion	O
microscopic	O
field	O
by	O
CD10	B-Gene_or_gene_product
staining	O
and	O
from	O
68	O
.	O
5	O
+	O
/	O
-	O
4	O
.	O
5	O
to	O
37	O
.	O
5	O
+	O
/	O
-	O
4	O
.	O
6	O
by	O
CD31	B-Gene_or_gene_product
staining	O
,	O
respectively	O
(	O
P	O
less	O
than	O
0	O
.	O
05	O
)	O
.	O

More	O
apoptotic	O
tumor	B-Cell
cells	I-Cell
were	O
seen	O
under	O
the	O
transmission	O
electron	O
microscope	O
.	O

CONCLUSION	O
:	O
Endostatin	B-Gene_or_gene_product
gene	O
therapy	O
mediated	O
by	O
adenoviral	O
vector	O
efficiently	O
induces	O
expression	O
of	O
endostatin	B-Gene_or_gene_product
in	O
vivo	O
,	O
and	O
inhibits	O
the	O
growth	O
and	O
metastasis	O
of	O
tumor	B-Cancer
.	O

It	O
is	O
concluded	O
that	O
its	O
action	O
is	O
targeted	O
to	O
tumor	B-Multi-tissue_structure
neovasculature	I-Multi-tissue_structure
and	O
the	O
mechanism	O
is	O
inhibition	O
of	O
tumor	B-Cancer
angiogenesis	O
.	O

The	O
role	O
of	O
p53	B-Gene_or_gene_product
in	O
pigmentation	O
,	O
tanning	O
and	O
melanoma	B-Cancer
.	O

p53	B-Gene_or_gene_product
has	O
a	O
central	O
role	O
in	O
skin	B-Organ
pigmentation	O
and	O
may	O
impact	O
on	O
melanoma	B-Cancer
at	O
all	O
stages	O
,	O
however	O
,	O
as	O
it	O
'	O
s	O
mutation	O
frequency	O
in	O
melanoma	B-Cancer
is	O
low	O
,	O
it	O
'	O
s	O
role	O
has	O
been	O
somewhat	O
under	O
-	O
appreciated	O
.	O

During	O
normal	O
skin	B-Organ
function	O
,	O
p53	B-Gene_or_gene_product
in	O
the	O
keratinocyte	B-Cell
is	O
a	O
transducer	O
of	O
the	O
skin	B-Organ
tanning	O
signal	O
and	O
an	O
essential	O
component	O
of	O
what	O
is	O
effectively	O
a	O
keratinocyte	B-Cell
-	O
melanocyte	B-Cell
signaling	O
cycle	O
that	O
regulates	O
skin	B-Organ
pigmentation	O
.	O

It	O
is	O
clear	O
that	O
this	O
cycle	O
functions	O
optimally	O
in	O
skin	B-Organ
of	O
dark	O
pigmentation	O
.	O

When	O
melanin	B-Simple_chemical
biosynthesis	O
is	O
genetically	O
disrupted	O
in	O
skin	B-Organ
of	O
white	B-Organism
complexion	I-Organism
,	O
we	O
propose	O
that	O
this	O
cycle	O
operates	O
as	O
a	O
promoter	O
of	O
melanocyte	B-Cell
proliferation	O
.	O

The	O
cell	B-Cell
autonomous	O
function	O
of	O
p53	B-Gene_or_gene_product
in	O
melanocytes	B-Cell
is	O
not	O
well	O
described	O
,	O
however	O
,	O
the	O
balance	O
of	O
the	O
evidence	O
suggests	O
that	O
p53	B-Gene_or_gene_product
is	O
an	O
effective	O
tumor	B-Cancer
suppressor	O
and	O
the	O
myriad	O
of	O
mechanisms	O
by	O
which	O
the	O
p53	B-Gene_or_gene_product
pathway	O
may	O
be	O
dysregulated	O
in	O
tumors	B-Cancer
attests	O
to	O
it	O
importance	O
as	O
a	O
tumor	B-Cancer
suppressor	O
.	O

In	O
this	O
review	O
,	O
we	O
outline	O
the	O
known	O
mechanisms	O
that	O
impair	O
p53	B-Gene_or_gene_product
itself	O
and	O
its	O
immediate	O
regulators	O
or	O
target	O
genes	O
during	O
melanomagenesis	O
.	O

Due	O
to	O
the	O
importance	O
of	O
this	O
pathway	O
,	O
it	O
is	O
clear	O
that	O
p53	B-Gene_or_gene_product
disruptions	O
may	O
relate	O
directly	O
to	O
a	O
patient	B-Organism
'	O
s	O
prognosis	O
.	O

This	O
pathway	O
will	O
continue	O
to	O
be	O
a	O
focus	O
of	O
investigation	O
,	O
particularly	O
with	O
respect	O
to	O
targeted	O
experimental	O
chemotherapeutics	O
.	O

Enhancement	O
of	O
docetaxel	B-Simple_chemical
-	O
induced	O
cytotoxicity	O
and	O
apoptosis	O
by	O
all	B-Simple_chemical
-	I-Simple_chemical
trans	I-Simple_chemical
retinoic	I-Simple_chemical
acid	I-Simple_chemical
(	O
ATRA	B-Simple_chemical
)	O
through	O
downregulation	O
of	O
survivin	B-Gene_or_gene_product
(	O
BIRC5	B-Gene_or_gene_product
)	O
,	O
MCL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
LTbeta	B-Gene_or_gene_product
-	I-Gene_or_gene_product
R	I-Gene_or_gene_product
in	O
hormone	O
-	O
and	O
drug	O
resistant	O
prostate	B-Cell
cancer	I-Cell
cell	I-Cell
line	I-Cell
,	O
DU	B-Cell
-	I-Cell
145	I-Cell
.	O

BACKGROUND	O
:	O
The	O
management	O
of	O
hormone	B-Cancer
-	I-Cancer
refractory	I-Cancer
prostate	I-Cancer
cancer	I-Cancer
(	O
HRPC	B-Cancer
)	O
still	O
remains	O
as	O
an	O
important	O
challenge	O
of	O
daily	O
oncology	O
practice	O
.	O

Docetaxel	B-Simple_chemical
has	O
proved	O
to	O
be	O
a	O
first	O
line	O
treatment	O
choice	O
.	O

All	B-Simple_chemical
-	I-Simple_chemical
trans	I-Simple_chemical
retinoic	I-Simple_chemical
acid	I-Simple_chemical
(	O
ATRA	B-Simple_chemical
)	O
could	O
potently	O
inhibit	O
the	O
growth	O
of	O
prostate	B-Cell
cancer	I-Cell
cells	I-Cell
in	O
vitro	O
and	O
its	O
combination	O
with	O
various	O
anticancer	B-Cancer
agents	O
results	O
in	O
increased	O
cytotoxicity	O
.	O

Based	O
on	O
these	O
data	O
,	O
our	O
aim	O
was	O
to	O
examine	O
the	O
synergistic	O
/	O
additive	O
cytotoxic	O
and	O
apoptotic	O
effects	O
of	O
combination	O
of	O
docetaxel	B-Simple_chemical
and	O
ATRA	B-Simple_chemical
,	O
in	O
hormone	O
-	O
and	O
drug	O
refractory	O
human	B-Organism
DU	B-Cell
-	I-Cell
145	I-Cell
prostate	I-Cell
cancer	I-Cell
cells	I-Cell
.	O

Furthermore	O
,	O
we	O
have	O
searched	O
for	O
the	O
underlying	O
mechanisms	O
of	O
apoptosis	O
by	O
demonstrating	O
apoptosis	O
-	O
related	O
genes	O
.	O

METHODS	O
:	O
XTT	B-Cell
cell	I-Cell
proliferation	O
assay	O
was	O
used	O
for	O
showing	O
cytotoxicity	O
.	O

For	O
verifying	O
apoptosis	O
,	O
both	O
DNA	B-Cellular_component
Fragmentation	O
by	O
ELISA	O
assay	O
and	O
caspase	B-Gene_or_gene_product
3	I-Gene_or_gene_product
/	O
7	B-Gene_or_gene_product
activity	O
measurement	O
were	O
used	O
.	O

For	O
detecting	O
the	O
mechanism	O
of	O
apoptosis	O
induced	O
by	O
docetaxel	B-Simple_chemical
-	O
ATRA	B-Simple_chemical
combination	O
,	O
OligoGeArray	O
which	O
consists	O
of	O
112	O
apoptosis	O
related	O
genes	O
was	O
used	O
.	O

RESULTS	O
:	O
Our	O
results	O
revealed	O
that	O
docetaxel	B-Simple_chemical
and	O
ATRA	B-Simple_chemical
were	O
synergistically	O
cytotoxic	O
and	O
apoptotic	O
in	O
DU	B-Cell
-	I-Cell
145	I-Cell
cells	I-Cell
,	O
in	O
a	O
dose	O
-	O
and	O
time	O
dependent	O
manner	O
.	O

It	O
was	O
also	O
shown	O
by	O
our	O
studies	O
that	O
apoptosis	O
was	O
induced	O
in	O
DU	B-Cell
-	I-Cell
145	I-Cell
prostate	I-Cell
carcinoma	I-Cell
cells	I-Cell
with	O
significant	O
cytotoxicity	O
,	O
no	O
matter	O
which	O
agent	O
applied	O
first	O
.	O

We	O
have	O
found	O
out	O
that	O
docetaxel	B-Simple_chemical
-	O
ATRA	B-Simple_chemical
combination	O
significantly	O
downregulates	O
survivin	B-Gene_or_gene_product
(	O
BIRC5	B-Gene_or_gene_product
)	O
,	O
myeloid	B-Gene_or_gene_product
cell	I-Gene_or_gene_product
leukemia	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
MCL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
and	O
lymphotoxin	B-Gene_or_gene_product
beta	I-Gene_or_gene_product
-	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
(	O
LTbetaR	B-Gene_or_gene_product
)	O
genes	O
,	O
which	O
all	O
three	O
have	O
pivotal	O
roles	O
in	O
regulation	O
of	O
apoptosis	O
and	O
cell	B-Cell
cycle	O
progression	O
.	O

CONCLUSION	O
:	O
In	O
conclusion	O
,	O
we	O
strongly	O
suggest	O
that	O
docetaxel	B-Simple_chemical
and	O
ATRA	B-Simple_chemical
combination	O
is	O
a	O
good	O
candidate	O
for	O
this	O
challenging	O
era	O
of	O
daily	O
oncologic	O
practice	O
.	O

Also	O
,	O
the	O
combination	O
of	O
docetaxel	B-Simple_chemical
and	O
ATRA	B-Simple_chemical
might	O
allow	O
a	O
reduction	O
in	O
docetaxel	B-Simple_chemical
doses	O
and	O
by	O
this	O
way	O
may	O
diminish	O
docetaxel	B-Simple_chemical
adverse	O
effects	O
while	O
maintaining	O
the	O
therapeutic	O
effect	O
in	O
patients	B-Organism
with	O
HRPC	B-Cancer
.	O

Diverse	O
cell	B-Cell
signaling	O
events	O
modulated	O
by	O
perlecan	B-Gene_or_gene_product
.	O

Perlecan	B-Gene_or_gene_product
is	O
a	O
ubiquitous	O
pericellular	B-Immaterial_anatomical_entity
proteoglycan	O
ideally	O
placed	O
to	O
mediate	O
cell	B-Cell
signaling	O
events	O
controlling	O
migration	O
,	O
proliferation	O
,	O
and	O
differentiation	O
.	O

Its	O
control	O
of	O
growth	O
factor	O
signaling	O
usually	O
involves	O
interactions	O
with	O
the	O
heparan	O
sulfate	O
chains	O
covalently	O
coupled	O
to	O
the	O
protein	O
core	O
'	O
s	O
N	O
-	O
terminus	O
.	O

However	O
,	O
this	O
modular	O
protein	O
core	O
also	O
binds	O
with	O
relatively	O
high	O
affinity	O
to	O
a	O
number	O
of	O
growth	O
factors	O
and	O
surface	O
receptors	O
,	O
thereby	O
stabilizing	O
cell	B-Cell
-	O
matrix	O
links	O
.	O

This	O
review	O
will	O
focus	O
on	O
perlecan	B-Gene_or_gene_product
-	O
growth	O
factor	O
interactions	O
and	O
describe	O
recent	O
advances	O
in	O
our	O
understanding	O
of	O
this	O
highly	O
conserved	O
proteoglycan	O
during	O
development	O
,	O
cancer	B-Cancer
growth	O
,	O
and	O
angiogenesis	O
.	O

The	O
pro	O
-	O
angiogenic	O
capacities	O
of	O
perlecan	B-Gene_or_gene_product
that	O
involve	O
proliferative	O
and	O
migratory	O
signals	O
in	O
response	O
to	O
bound	O
growth	O
factors	O
will	O
be	O
explored	O
,	O
as	O
well	O
as	O
the	O
anti	O
-	O
angiogenic	O
signals	O
resulting	O
from	O
interactions	O
between	O
the	O
C	O
-	O
terminal	O
domain	O
known	O
as	O
endorepellin	B-Gene_or_gene_product
and	O
integrins	B-Gene_or_gene_product
that	O
control	O
adhesion	O
of	O
cells	B-Cell
to	O
the	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
.	O

These	O
two	O
somewhat	O
diametrically	O
opposed	O
roles	O
will	O
be	O
discussed	O
in	O
light	O
of	O
new	O
data	O
emerging	O
from	O
various	O
fields	O
which	O
converge	O
on	O
perlecan	B-Gene_or_gene_product
as	O
a	O
key	O
regulator	O
of	O
cell	B-Cell
growth	O
and	O
angiogenesis	O
.	O

Activation	O
of	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
through	O
a	O
phosphomimetic	O
serine	B-Amino_acid
727	I-Amino_acid
promotes	O
prostate	B-Organ
tumorigenesis	O
independent	O
of	O
tyrosine	B-Amino_acid
705	I-Amino_acid
phosphorylation	O
.	O

Aberrantly	O
activated	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
is	O
implicated	O
in	O
the	O
development	O
of	O
various	O
human	B-Organism
cancers	B-Cancer
.	O

Y705	B-Amino_acid
phosphorylation	O
is	O
conventionally	O
thought	O
to	O
be	O
required	O
for	O
Stat3	B-Gene_or_gene_product
signal	O
-	O
dependent	O
activation	O
and	O
seems	O
to	O
play	O
an	O
essential	O
role	O
in	O
some	O
malignancies	B-Cancer
.	O

Recently	O
,	O
it	O
was	O
shown	O
that	O
Stat3	B-Gene_or_gene_product
is	O
activated	O
through	O
novel	O
and	O
noncanonical	O
mechanisms	O
,	O
including	O
phosphorylation	O
at	O
S727	B-Amino_acid
.	O

Here	O
,	O
we	O
investigate	O
S727	B-Amino_acid
phosphorylation	O
of	O
Stat3	B-Gene_or_gene_product
and	O
its	O
subsequent	O
effects	O
in	O
prostate	B-Cancer
cancer	I-Cancer
development	O
,	O
independent	O
of	O
Y705	B-Amino_acid
phosphorylation	O
,	O
using	O
mutated	O
Stat3	B-Gene_or_gene_product
in	O
the	O
human	B-Organism
prostate	B-Cell
cancer	I-Cell
cell	I-Cell
line	I-Cell
LNCaP	I-Cell
.	O

We	O
show	O
mutation	O
of	O
S727	B-Amino_acid
to	O
the	O
phosphomimetic	O
residue	O
Glu	B-Amino_acid
,	O
and	O
inactivation	O
of	O
Y705	B-Amino_acid
(	O
Y705F	B-Amino_acid
/	O
S727E	B-Amino_acid
)	O
resulted	O
in	O
a	O
remarkable	O
growth	O
advantage	O
in	O
low	O
-	O
serum	B-Organism_substance
,	O
enhanced	O
anchorage	O
-	O
independent	O
growth	O
in	O
soft	O
agar	O
,	O
and	O
increased	O
tumorigenicity	O
in	O
nonobese	B-Organism
diabetic	I-Organism
/	I-Organism
severe	I-Organism
combined	I-Organism
immunodeficient	I-Organism
(	I-Organism
NOD	I-Organism
/	I-Organism
SCID	I-Organism
)	I-Organism
mice	I-Organism
,	O
possibly	O
by	O
direct	O
activation	O
of	O
downstream	O
proto	O
-	O
oncogenes	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
,	O
mcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
and	O
survivin	B-Gene_or_gene_product
.	O

Y705F	B-Amino_acid
/	O
S727E	B-Amino_acid
mutant	O
cells	O
were	O
more	O
invasive	O
than	O
Y705F	B-Amino_acid
/	O
S727A	B-Amino_acid
(	O
inactivation	O
of	O
Y705	B-Amino_acid
and	O
S727	B-Amino_acid
)	O
mutant	O
cells	O
,	O
and	O
more	O
Y705F	B-Amino_acid
/	O
S727E	B-Amino_acid
mutant	O
Stat3	B-Gene_or_gene_product
was	O
localized	O
in	O
the	O
nuclei	B-Cellular_component
relative	O
to	O
Y705F	B-Amino_acid
/	O
S727A	B-Amino_acid
mutant	O
Stat3	B-Gene_or_gene_product
at	O
the	O
steady	O
state	O
.	O

Furthermore	O
,	O
the	O
Y705F	B-Amino_acid
/	O
S727E	B-Amino_acid
but	O
not	O
the	O
Y705F	B-Amino_acid
/	O
S727A	B-Amino_acid
mutant	O
induced	O
anchorage	O
-	O
independent	O
growth	O
of	O
noncancerous	B-Cell
prostate	I-Cell
epithelial	I-Cell
cells	I-Cell
(	O
RWPE	B-Cell
-	I-Cell
1	I-Cell
)	O
.	O

We	O
further	O
show	O
that	O
Stat3	B-Gene_or_gene_product
is	O
phosphorylated	O
at	O
S727	B-Amino_acid
in	O
65	O
%	O
of	O
malignant	B-Tissue
prostate	I-Tissue
tissues	I-Tissue
(	O
n	O
=	O
20	O
)	O
relative	O
to	O
25	O
%	O
of	O
normal	B-Tissue
prostate	I-Tissue
tissues	I-Tissue
(	O
n	O
=	O
4	O
)	O
.	O

Moreover	O
,	O
there	O
is	O
a	O
positive	O
correlation	O
between	O
phosphoS727	B-Amino_acid
-	O
Stat3	B-Gene_or_gene_product
expression	O
and	O
Gleason	O
score	O
in	O
these	O
prostate	B-Tissue
cancer	I-Tissue
tissues	I-Tissue
(	O
P	O
=	O
0	O
.	O
05	O
)	O
.	O

Our	O
data	O
suggest	O
for	O
the	O
first	O
time	O
that	O
S727	B-Amino_acid
phosphorylation	O
is	O
sufficient	O
to	O
activate	O
Stat3	B-Gene_or_gene_product
,	O
thereby	O
driving	O
prostate	B-Organ
tumorigenesis	O
independent	O
of	O
Y705	B-Amino_acid
phosphorylation	O
.	O

Allergic	O
pulmonary	B-Organ
inflammation	O
promotes	O
the	O
recruitment	O
of	O
circulating	O
tumor	B-Cell
cells	I-Cell
to	O
the	O
lung	B-Organ
.	O

Allergen	O
-	O
induced	O
respiratory	B-Anatomical_system
inflammation	O
facilitates	O
and	O
/	O
or	O
elicits	O
the	O
extravasation	O
of	O
proinflammatory	B-Cell
leukocytes	I-Cell
by	O
well	O
-	O
understood	O
mechanisms	O
that	O
mediate	O
the	O
movement	O
of	O
multiple	O
cell	B-Cell
types	O
.	O

The	O
nonspecific	O
character	O
of	O
these	O
pathways	O
led	O
us	O
to	O
hypothesize	O
that	O
circulating	O
cancer	B-Cell
cells	I-Cell
use	O
similar	O
mechanisms	O
,	O
promoting	O
secondary	B-Cancer
tumor	I-Cancer
formation	O
at	O
distal	O
sites	O
.	O

To	O
test	O
this	O
hypothesis	O
,	O
the	O
frequency	O
of	O
metastasis	O
to	O
the	O
lung	B-Organ
as	O
a	O
function	O
of	O
allergic	O
pulmonary	B-Organ
inflammation	O
was	O
assessed	O
following	O
the	O
i	B-Immaterial_anatomical_entity
.	I-Immaterial_anatomical_entity
v	I-Immaterial_anatomical_entity
.	I-Immaterial_anatomical_entity
injection	O
of	O
B16	B-Cell
-	I-Cell
F10	I-Cell
melanoma	I-Cell
cells	I-Cell
in	O
mice	B-Organism
.	O

These	O
studies	O
showed	O
that	O
allergen	O
-	O
induced	O
pulmonary	B-Organ
inflammation	O
resulted	O
in	O
a	O
>	O
3	O
-	O
fold	O
increase	O
in	O
lung	B-Organ
metastases	O
.	O

This	O
increase	O
was	O
dependent	O
on	O
CD4	B-Gene_or_gene_product
(	O
+	O
)	O
T	O
-	O
cell	O
activities	O
;	O
however	O
,	O
it	O
occurred	O
independent	O
of	O
the	O
induced	O
eosinophilia	O
associated	O
with	O
allergen	O
provocation	O
.	O

Interventional	O
strategies	O
showed	O
that	O
existing	O
therapeutic	O
modalities	O
for	O
asthma	O
,	O
such	O
as	O
inhaled	O
corticosteroids	B-Simple_chemical
,	O
were	O
sufficient	O
to	O
block	O
the	O
enhanced	O
pulmonary	B-Organ
recruitment	O
of	O
cancer	B-Cell
cells	I-Cell
from	O
circulation	O
.	O

Additional	O
mechanistic	O
studies	O
further	O
suggested	O
that	O
the	O
ability	O
of	O
circulating	O
cancer	B-Cell
cells	I-Cell
to	O
extravasate	O
to	O
surrounding	O
lung	B-Tissue
tissues	I-Tissue
was	O
linked	O
to	O
the	O
activation	O
of	O
the	O
vascular	B-Tissue
endothelium	I-Tissue
via	O
one	O
or	O
more	O
Galpha	B-Gene_or_gene_product
(	I-Gene_or_gene_product
i	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	O
coupled	O
receptors	O
.	O

Interestingly	O
,	O
a	O
survey	O
of	O
a	O
clinical	O
breast	B-Cancer
cancer	I-Cancer
surgical	O
database	O
showed	O
that	O
the	O
incidence	O
of	O
asthma	O
was	O
higher	O
among	O
patients	B-Organism
with	O
lung	B-Cancer
metastases	I-Cancer
.	O

Thus	O
,	O
our	O
data	O
show	O
that	O
allergic	O
respiratory	B-Anatomical_system
inflammation	O
may	O
represent	O
a	O
risk	O
factor	O
for	O
the	O
development	O
of	O
lung	B-Cancer
metastases	I-Cancer
and	O
suggest	O
that	O
amelioration	O
of	O
the	O
pulmonary	B-Organ
inflammation	O
associated	O
with	O
asthma	O
will	O
have	O
a	O
direct	O
and	O
immediate	O
benefit	O
to	O
the	O
7	O
%	O
to	O
8	O
%	O
of	O
breast	B-Cancer
cancer	I-Cancer
patients	B-Organism
with	O
this	O
lung	B-Organ
disease	O
.	O

Inhibition	O
of	O
energy	O
-	O
producing	O
pathways	O
of	O
HepG2	B-Cell
cells	I-Cell
by	O
3	B-Simple_chemical
-	I-Simple_chemical
bromopyruvate	I-Simple_chemical
.	O

3	B-Simple_chemical
-	I-Simple_chemical
BrPA	I-Simple_chemical
(	O
3	B-Simple_chemical
-	I-Simple_chemical
bromopyruvate	I-Simple_chemical
)	O
is	O
an	O
alkylating	O
agent	O
with	O
anti	O
-	O
tumoral	B-Cancer
activity	O
on	O
hepatocellular	B-Cancer
carcinoma	I-Cancer
.	O

This	O
compound	O
inhibits	O
cellular	B-Cell
ATP	B-Simple_chemical
production	O
owing	O
to	O
its	O
action	O
on	O
glycolysis	O
and	O
oxidative	O
phosphorylation	O
;	O
however	O
,	O
the	O
specific	O
metabolic	O
steps	O
and	O
mechanisms	O
of	O
3	B-Simple_chemical
-	I-Simple_chemical
BrPA	I-Simple_chemical
action	O
in	O
human	B-Organism
hepatocellular	B-Cancer
carcinomas	I-Cancer
,	O
particularly	O
its	O
effects	O
on	O
mitochondrial	B-Cellular_component
energetics	O
,	O
are	O
poorly	O
understood	O
.	O

In	O
the	O
present	O
study	O
it	O
was	O
found	O
that	O
incubation	O
of	O
HepG2	B-Cell
cells	I-Cell
with	O
a	O
low	O
concentration	O
of	O
3	B-Simple_chemical
-	I-Simple_chemical
BrPA	I-Simple_chemical
for	O
a	O
short	O
period	O
(	O
150	O
microM	O
for	O
30	O
min	O
)	O
significantly	O
affected	O
both	O
glycolysis	O
and	O
mitochondrial	B-Cellular_component
respiratory	O
functions	O
.	O

The	O
activity	O
of	O
mitochondrial	B-Cellular_component
hexokinase	B-Gene_or_gene_product
was	O
not	O
inhibited	O
by	O
150	O
microM	O
3	B-Simple_chemical
-	I-Simple_chemical
BrPA	I-Simple_chemical
,	O
but	O
this	O
concentration	O
caused	O
more	O
than	O
70	O
%	O
inhibition	O
of	O
GAPDH	B-Gene_or_gene_product
(	O
glyceraldehyde	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
-	I-Gene_or_gene_product
phosphate	I-Gene_or_gene_product
dehydrogenase	I-Gene_or_gene_product
)	O
and	O
3	B-Gene_or_gene_product
-	I-Gene_or_gene_product
phosphoglycerate	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
activities	O
.	O

Additionally	O
,	O
3	B-Simple_chemical
-	I-Simple_chemical
BrPA	I-Simple_chemical
treatment	O
significantly	O
impaired	O
lactate	B-Simple_chemical
production	O
by	O
HepG2	B-Cell
cells	I-Cell
,	O
even	O
when	O
glucose	B-Simple_chemical
was	O
withdrawn	O
from	O
the	O
incubation	O
medium	O
.	O

Oxygen	B-Simple_chemical
consumption	O
of	O
HepG2	B-Cell
cells	I-Cell
supported	O
by	O
either	O
pyruvate	B-Simple_chemical
/	O
malate	B-Simple_chemical
or	O
succinate	B-Simple_chemical
was	O
inhibited	O
when	O
cells	B-Cell
were	O
pre	O
-	O
incubated	O
with	O
3	B-Simple_chemical
-	I-Simple_chemical
BrPA	I-Simple_chemical
in	O
glucose	B-Simple_chemical
-	O
free	O
medium	O
.	O

On	O
the	O
other	O
hand	O
,	O
when	O
cells	B-Cell
were	O
pre	O
-	O
incubated	O
in	O
glucose	B-Simple_chemical
-	O
supplemented	O
medium	O
,	O
oxygen	B-Simple_chemical
consumption	O
was	O
affected	O
only	O
when	O
succinate	B-Simple_chemical
was	O
used	O
as	O
the	O
oxidizable	O
substrate	O
.	O

An	O
increase	O
in	O
oligomycin	B-Simple_chemical
-	O
independent	O
respiration	O
was	O
observed	O
in	O
HepG2	B-Cell
cells	I-Cell
treated	O
with	O
3	B-Simple_chemical
-	I-Simple_chemical
BrPA	I-Simple_chemical
only	O
when	O
incubated	O
in	O
glucose	B-Simple_chemical
-	O
supplemented	O
medium	O
,	O
indicating	O
that	O
3	B-Simple_chemical
-	I-Simple_chemical
BrPA	I-Simple_chemical
induces	O
mitochondrial	B-Cellular_component
proton	B-Simple_chemical
leakage	O
as	O
well	O
as	O
blocking	O
the	O
electron	B-Simple_chemical
transport	O
system	O
.	O

The	O
activity	O
of	O
succinate	B-Gene_or_gene_product
dehydrogenase	I-Gene_or_gene_product
was	O
inhibited	O
by	O
70	O
%	O
by	O
3	B-Simple_chemical
-	I-Simple_chemical
BrPA	I-Simple_chemical
treatment	O
.	O

These	O
results	O
suggest	O
that	O
the	O
combined	O
action	O
of	O
3	B-Simple_chemical
-	I-Simple_chemical
BrPA	I-Simple_chemical
on	O
succinate	B-Gene_or_gene_product
dehydrogenase	I-Gene_or_gene_product
and	O
on	O
glycolysis	O
,	O
inhibiting	O
steps	O
downstream	O
of	O
the	O
phosphorylation	O
of	O
glucose	B-Simple_chemical
,	O
play	O
an	O
important	O
role	O
in	O
HepG2	B-Cell
cell	I-Cell
death	O
.	O

[	O
Retinochoroidopathy	O
after	O
intravitreal	O
anti	O
-	O
VEGF	B-Gene_or_gene_product
treatment	O
]	O

A	O
74	O
-	O
year	O
-	O
old	O
man	B-Organism
presented	O
with	O
persistent	O
metamorphopsias	O
of	O
the	O
right	O
eye	B-Organ
2	O
weeks	O
after	O
intravitreal	O
injection	O
of	O
bevacizumab	B-Simple_chemical
to	O
treat	O
choroidal	B-Multi-tissue_structure
neovascularization	O
due	O
to	O
exudative	O
age	O
-	O
related	O
macular	B-Tissue
degeneration	O
.	O

The	O
diagnosis	O
reached	O
was	O
retinochoroiditis	O
as	O
an	O
occult	O
manifestation	O
of	O
sarcoidosis	O
,	O
possibly	O
resulting	O
from	O
an	O
intravitreal	O
injection	O
of	O
bevacizumab	B-Simple_chemical
.	O

The	O
patient	B-Organism
received	O
a	O
prescription	O
for	O
100	O
mg	O
Ultralan	B-Simple_chemical
to	O
be	O
taken	O
daily	O
for	O
3	O
days	O
and	O
then	O
tapered	O
in	O
3	O
day	O
steps	O
.	O

During	O
the	O
further	O
course	O
no	O
deterioration	O
of	O
the	O
condition	O
was	O
observed	O
.	O

Silencing	O
of	O
directional	O
migration	O
in	O
roundabout4	B-Gene_or_gene_product
knockdown	O
endothelial	O
cells	O
.	O

BACKGROUND	O
:	O
Roundabouts	B-Gene_or_gene_product
are	O
axon	O
guidance	O
molecules	O
that	O
have	O
recently	O
been	O
identified	O
to	O
play	O
a	O
role	O
in	O
vascular	B-Multi-tissue_structure
guidance	O
as	O
well	O
.	O

In	O
this	O
study	O
,	O
we	O
have	O
investigated	O
gene	O
knockdown	O
analysis	O
of	O
endothelial	B-Cell
Robos	B-Gene_or_gene_product
,	O
in	O
particular	O
roundabout	B-Gene_or_gene_product
4	I-Gene_or_gene_product
(	O
robo4	B-Gene_or_gene_product
)	O
,	O
the	O
predominant	O
Robo	B-Gene_or_gene_product
in	O
endothelial	B-Cell
cells	I-Cell
using	O
small	O
interfering	O
RNA	O
technology	O
in	O
vitro	O
.	O

RESULTS	O
:	O
Robo1	B-Gene_or_gene_product
and	O
Robo4	B-Gene_or_gene_product
knockdown	O
cells	O
display	O
distinct	O
activity	O
in	O
endothelial	B-Cell
cell	I-Cell
migration	O
assay	O
.	O

The	O
knockdown	O
of	O
robo4	B-Gene_or_gene_product
abrogated	O
the	O
chemotactic	O
response	O
of	O
endothelial	B-Cell
cells	I-Cell
to	O
serum	B-Organism_substance
but	O
enhanced	O
a	O
chemokinetic	O
response	O
to	O
Slit2	B-Gene_or_gene_product
,	O
while	O
robo1	B-Gene_or_gene_product
knockdown	O
cells	O
do	O
not	O
display	O
chemotactic	O
response	O
to	O
serum	B-Organism_substance
or	O
VEGF	B-Gene_or_gene_product
.	O

Robo4	B-Gene_or_gene_product
knockdown	O
endothelial	O
cells	O
unexpectedly	O
show	O
up	O
regulation	O
of	O
Rho	B-Gene_or_gene_product
GTPases	I-Gene_or_gene_product
.	O

Zebrafish	B-Organism
Robo4	B-Gene_or_gene_product
rescues	O
both	O
Rho	B-Gene_or_gene_product
GTPase	I-Gene_or_gene_product
homeostasis	O
and	O
serum	B-Organism_substance
reduced	O
chemotaxis	O
in	O
robo4	B-Gene_or_gene_product
knockdown	O
cells	O
.	O

Robo1	B-Gene_or_gene_product
and	O
Robo4	B-Gene_or_gene_product
interact	O
and	O
share	O
molecules	O
such	O
as	O
Slit2	B-Gene_or_gene_product
,	O
Mena	B-Gene_or_gene_product
and	O
Vilse	B-Gene_or_gene_product
,	O
a	O
Cdc42	B-Gene_or_gene_product
-	I-Gene_or_gene_product
GAP	I-Gene_or_gene_product
.	O

In	O
addition	O
,	O
this	O
study	O
mechanistically	O
implicates	O
IRSp53	B-Gene_or_gene_product
in	O
the	O
signaling	O
nexus	O
between	O
activated	O
Cdc42	B-Gene_or_gene_product
and	O
Mena	B-Gene_or_gene_product
,	O
both	O
of	O
which	O
have	O
previously	O
been	O
shown	O
to	O
be	O
involved	O
with	O
Robo4	B-Gene_or_gene_product
signaling	O
in	O
endothelial	B-Cell
cells	I-Cell
.	O

CONCLUSION	O
:	O
This	O
study	O
identifies	O
specific	O
components	O
of	O
the	O
Robo	B-Gene_or_gene_product
signaling	O
apparatus	O
that	O
work	O
together	O
to	O
guide	O
directional	O
migration	O
of	O
endothelial	B-Cell
cells	I-Cell
.	O

p53	B-Gene_or_gene_product
hot	O
-	O
spot	O
mutants	O
increase	O
tumor	B-Cancer
vascularization	O
via	O
ROS	B-Simple_chemical
-	O
mediated	O
activation	O
of	O
the	O
HIF1	B-Gene_or_gene_product
/	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
A	I-Gene_or_gene_product
pathway	O
.	O

The	O
function	O
of	O
p53	B-Gene_or_gene_product
tumor	B-Cancer
suppressor	O
is	O
often	O
altered	O
in	O
various	O
human	B-Organism
tumors	B-Cancer
predominantly	O
through	O
missense	O
-	O
mutations	O
resulting	O
in	O
accumulation	O
of	O
mutant	O
proteins	O
.	O

We	O
revealed	O
that	O
expression	O
of	O
p53	B-Gene_or_gene_product
proteins	O
with	O
amino	O
-	O
acid	O
substitutions	O
at	O
codons	O
175	O
(	O
R175H	O
)	O
,	O
248	O
(	O
R248W	O
)	O
,	O
and	O
273	O
(	O
R273H	O
)	O
,	O
representing	O
the	O
hot	O
-	O
spots	O
of	O
mutations	O
in	O
various	O
human	B-Organism
tumors	B-Cancer
,	O
increased	O
the	O
number	O
of	O
vessels	B-Multi-tissue_structure
in	O
HCT116	B-Cancer
human	I-Cancer
colon	I-Cancer
carcinoma	I-Cancer
xenografts	I-Cancer
and	O
,	O
as	O
a	O
result	O
,	O
accelerated	O
their	O
growth	O
.	O

Stimulation	O
of	O
tumor	B-Cancer
angiogenesis	O
was	O
connected	O
with	O
about	O
2	O
-	O
fold	O
increase	O
in	O
intracellular	B-Immaterial_anatomical_entity
level	O
of	O
reactive	B-Simple_chemical
oxygen	I-Simple_chemical
species	I-Simple_chemical
(	O
ROS	B-Simple_chemical
)	O
.	O

Antioxidant	O
N	B-Simple_chemical
-	I-Simple_chemical
acetyl	I-Simple_chemical
-	I-Simple_chemical
l	I-Simple_chemical
-	I-Simple_chemical
aspartate	I-Simple_chemical
(	O
NAC	B-Simple_chemical
)	O
decreased	O
vessels	B-Multi-tissue_structure
number	O
in	O
tumors	B-Cancer
formed	O
by	O
cells	B-Cell
with	O
inactivated	O
p53	B-Gene_or_gene_product
and	O
inhibited	O
their	O
growth	O
.	O

Effect	O
of	O
ROS	B-Simple_chemical
on	O
angiogenesis	O
in	O
tumors	B-Cancer
expressing	O
hot	O
-	O
spot	O
p53	B-Gene_or_gene_product
mutants	O
was	O
correlated	O
with	O
their	O
ability	O
to	O
increase	O
a	O
content	O
of	O
HIF1	B-Gene_or_gene_product
transcriptional	O
factor	O
responsible	O
for	O
up	O
-	O
regulation	O
of	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
A	I-Gene_or_gene_product
mRNAs	O
.	O

The	O
Akt	B-Gene_or_gene_product
/	O
mTOR	B-Gene_or_gene_product
pathway	O
assures	O
the	O
synthesis	O
of	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
protein	O
in	O
a	O
glucose	B-Simple_chemical
-	O
and	O
reoxygenation	O
-	O
dependent	O
manner	O
in	O
irradiated	O
tumors	B-Cancer
.	O

Transcriptional	O
activity	O
of	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-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
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
has	O
been	O
reported	O
to	O
be	O
up	O
-	O
regulated	O
in	O
solid	B-Cancer
tumors	I-Cancer
after	O
ionizing	O
radiation	O
;	O
however	O
,	O
the	O
molecular	O
mechanism	O
underlying	O
the	O
response	O
remains	O
to	O
be	O
elucidated	O
.	O

In	O
the	O
present	O
study	O
,	O
we	O
performed	O
a	O
series	O
of	O
molecular	O
imaging	O
experiments	O
using	O
a	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
dependent	O
reporter	O
gene	O
,	O
5HREp	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ODD	I-Gene_or_gene_product
-	I-Gene_or_gene_product
luc	I-Gene_or_gene_product
,	O
and	O
found	O
an	O
essential	O
role	O
of	O
the	O
Akt	B-Gene_or_gene_product
/	O
mTOR	B-Gene_or_gene_product
pathway	O
.	O

Hypoxic	O
tumor	B-Cell
cells	I-Cell
distant	O
from	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
were	O
dramatically	O
reoxygenated	O
at	O
24	O
h	O
postirradiation	O
,	O
and	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
activity	O
increased	O
as	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
accumulated	O
in	O
the	O
reoxygenated	O
regions	O
.	O

The	O
accumulation	O
was	O
inhibited	O
with	O
a	O
nonmetabolizable	O
glucose	B-Simple_chemical
analog	O
,	O
2	B-Simple_chemical
-	I-Simple_chemical
deoxy	I-Simple_chemical
-	I-Simple_chemical
d	I-Simple_chemical
-	I-Simple_chemical
glucose	I-Simple_chemical
,	O
through	O
the	O
suppression	O
of	O
radiation	O
-	O
induced	O
phosphorylation	O
of	O
Akt	B-Gene_or_gene_product
in	O
the	O
reoxygenated	O
regions	O
.	O

Akt	B-Gene_or_gene_product
knockdown	O
and	O
an	O
mTOR	B-Gene_or_gene_product
inhibitor	O
revealed	O
the	O
importance	O
of	O
the	O
Akt	B-Gene_or_gene_product
/	O
mTOR	B-Gene_or_gene_product
pathway	O
in	O
the	O
postirradiation	O
accumulation	O
of	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
.	O

In	O
vitro	O
experiments	O
confirmed	O
that	O
an	O
increase	O
in	O
glucose	B-Simple_chemical
availability	O
induced	O
Akt	B-Gene_or_gene_product
phosphorylation	O
under	O
reoxygenated	O
conditions	O
and	O
consequently	O
up	O
-	O
regulated	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
translation	O
.	O

Moreover	O
,	O
both	O
the	O
accelerated	O
translation	O
and	O
the	O
previously	O
reported	O
reactive	B-Simple_chemical
oxygen	I-Simple_chemical
species	I-Simple_chemical
-	O
mediated	O
stabilization	O
of	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
protein	O
were	O
essential	O
to	O
the	O
activation	O
of	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

All	O
of	O
these	O
results	O
indicate	O
that	O
Akt	B-Gene_or_gene_product
/	O
mTOR	B-Gene_or_gene_product
-	O
dependent	O
translation	O
of	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
plays	O
a	O
critical	O
role	O
in	O
the	O
postirradiation	O
up	O
-	O
regulation	O
of	O
intratumoral	B-Cancer
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
activity	O
in	O
response	O
to	O
radiation	O
-	O
induced	O
alterations	O
of	O
glucose	B-Simple_chemical
and	O
oxygen	B-Simple_chemical
availability	O
in	O
a	O
solid	B-Cancer
tumor	I-Cancer
.	O

Estradiol	B-Simple_chemical
increases	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
secretion	O
of	O
normal	O
human	B-Organism
breast	B-Tissue
tissue	I-Tissue
and	O
breast	B-Cancer
cancer	I-Cancer
in	O
vivo	O
.	O

IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
or	O
CXCL8	B-Gene_or_gene_product
has	O
been	O
associated	O
with	O
tumor	B-Cancer
angiogenesis	O
,	O
metastasis	O
,	O
and	O
poor	O
prognosis	O
in	O
breast	B-Cancer
cancer	I-Cancer
.	O

Estrogen	B-Simple_chemical
is	O
crucial	O
in	O
breast	B-Organism_subdivision
carcinogenesis	O
and	O
tumor	B-Cancer
progression	O
.	O

Whether	O
sex	O
steroids	O
affect	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
secretion	O
of	O
normal	O
breast	B-Tissue
tissue	I-Tissue
or	O
breast	B-Cancer
cancer	I-Cancer
is	O
not	O
known	O
.	O

Several	O
cell	B-Cell
types	O
in	O
a	O
tissue	B-Tissue
secrete	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
.	O

Hence	O
,	O
regulatory	O
mechanisms	O
of	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
need	O
to	O
be	O
investigated	O
in	O
whole	O
tissue	B-Tissue
.	O

We	O
used	O
microdialysis	O
to	O
sample	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
in	O
normal	O
human	B-Organism
breast	B-Tissue
tissue	I-Tissue
in	O
situ	O
in	O
pre	O
-	O
and	O
postmenopausal	O
women	B-Organism
,	O
preoperatively	O
in	O
breast	B-Cancer
cancers	I-Cancer
of	O
women	B-Organism
,	O
and	O
in	O
experimental	O
breast	B-Cancer
cancer	I-Cancer
in	O
mice	B-Organism
.	O

We	O
found	O
a	O
significant	O
positive	O
correlation	O
between	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
and	O
estradiol	B-Simple_chemical
in	O
normal	O
breast	B-Tissue
tissue	I-Tissue
and	O
hormone	B-Cancer
-	I-Cancer
dependent	I-Cancer
breast	I-Cancer
cancer	I-Cancer
in	O
vivo	O
.	O

Ex	O
vivo	O
,	O
estradiol	B-Simple_chemical
exposure	O
increased	O
the	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
secretion	O
of	O
normal	O
whole	O
breast	B-Tissue
tissue	I-Tissue
in	O
culture	O
.	O

In	O
experimental	O
breast	B-Cancer
cancer	I-Cancer
,	O
estradiol	B-Simple_chemical
increased	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
whereas	O
the	O
anti	O
-	O
estrogen	B-Simple_chemical
tamoxifen	B-Simple_chemical
inhibited	O
the	O
secretion	O
of	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
both	O
in	O
vitro	O
and	O
extracellularly	O
in	O
vivo	O
in	O
tumors	B-Cancer
of	O
nude	B-Organism
mice	I-Organism
.	O

An	O
anti	B-Simple_chemical
-	I-Simple_chemical
IL	I-Simple_chemical
-	I-Simple_chemical
8	I-Simple_chemical
Ab	I-Simple_chemical
inhibited	O
endothelial	B-Cell
cell	I-Cell
proliferation	O
induced	O
by	O
cancer	B-Cell
cell	I-Cell
produced	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
and	O
tumors	B-Cancer
with	O
low	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
levels	O
exhibited	O
decreased	O
angiogenesis	O
.	O

Our	O
results	O
strongly	O
suggest	O
that	O
estradiol	B-Simple_chemical
has	O
a	O
critical	O
role	O
in	O
the	O
regulation	O
of	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
in	O
normal	O
human	B-Organism
breast	B-Tissue
tissue	I-Tissue
and	O
human	B-Organism
breast	B-Cancer
cancer	I-Cancer
.	O

IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
may	O
present	O
a	O
novel	O
therapeutic	O
target	O
for	O
estrogen	B-Simple_chemical
driven	O
breast	B-Organism_subdivision
carcinogenesis	O
and	O
tumor	B-Cancer
progression	O
.	O

Chemokine	O
receptor	O
CXCR4	B-Gene_or_gene_product
expression	O
,	O
function	O
,	O
and	O
clinical	O
implications	O
in	O
gastric	B-Cancer
cancer	I-Cancer
.	O

The	O
chemokine	O
receptor	O
CXCR4	B-Gene_or_gene_product
is	O
associated	O
with	O
the	O
biological	O
behavior	O
of	O
cancer	B-Cancer
,	O
but	O
few	O
studies	O
have	O
addressed	O
the	O
expression	O
and	O
function	O
of	O
CXCR4	B-Gene_or_gene_product
in	O
human	B-Organism
gastric	B-Cancer
cancer	I-Cancer
and	O
its	O
impact	O
on	O
disease	O
prognosis	O
.	O

We	O
studied	O
the	O
expression	O
of	O
CXCR4	B-Gene_or_gene_product
using	O
RT	O
-	O
PCR	O
,	O
Western	O
blotting	O
,	O
flow	O
cytometry	O
,	O
and	O
confocal	O
microscopy	O
in	O
five	O
gastric	B-Cell
cancer	I-Cell
cell	I-Cell
lines	I-Cell
.	O

We	O
also	O
examined	O
cell	B-Cell
proliferation	O
,	O
migration	O
,	O
and	O
anti	O
-	O
apoptotic	O
activity	O
in	O
response	O
to	O
stromal	B-Gene_or_gene_product
cell	I-Gene_or_gene_product
-	I-Gene_or_gene_product
derived	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	I-Gene_or_gene_product
SDF	I-Gene_or_gene_product
)	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
and	O
evaluated	O
SDF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
/	O
CXCR4	B-Gene_or_gene_product
signaling	O
pathways	O
.	O

Furthermore	O
,	O
we	O
investigated	O
the	O
correlation	O
between	O
CXCR4	B-Gene_or_gene_product
expression	O
and	O
the	O
clinical	O
features	O
of	O
221	O
gastric	B-Tissue
cancer	I-Tissue
tissue	I-Tissue
samples	I-Tissue
.	O

CXCR4	B-Gene_or_gene_product
transcripts	O
and	O
proteins	O
were	O
detectable	O
in	O
all	O
five	O
gastric	B-Cell
cancer	I-Cell
cell	I-Cell
lines	I-Cell
.	O

However	O
,	O
MKN	B-Cell
-	I-Cell
28	I-Cell
,	O
MKN	B-Cell
-	I-Cell
45	I-Cell
,	O
MKN	B-Cell
-	I-Cell
74	I-Cell
,	O
and	O
SNU16	B-Cell
cells	I-Cell
did	O
not	O
express	O
membrane	B-Cellular_component
CXCR4	B-Gene_or_gene_product
.	O

In	O
contrast	O
,	O
KATO	B-Cell
III	I-Cell
cells	I-Cell
expressed	O
membrane	B-Cellular_component
CXCR4	B-Gene_or_gene_product
.	O

In	O
these	O
cells	B-Cell
,	O
SDF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
-	O
induced	O
migration	O
was	O
observed	O
and	O
was	O
blocked	O
by	O
AMD3100	B-Simple_chemical
,	O
a	O
specific	O
inhibitor	O
of	O
CXCR4	B-Gene_or_gene_product
.	O

SDF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
induced	O
rapid	O
phosphorylation	O
of	O
Erk1	B-Gene_or_gene_product
/	I-Gene_or_gene_product
2	I-Gene_or_gene_product
MAPK	I-Gene_or_gene_product
but	O
did	O
not	O
promote	O
phosphorylation	O
of	O
Stat3	B-Gene_or_gene_product
or	O
Akt	B-Gene_or_gene_product
.	O

Gastric	B-Tissue
cancer	I-Tissue
tissue	I-Tissue
samples	I-Tissue
expressed	O
CXCR4	B-Gene_or_gene_product
with	O
variable	O
intensities	O
.	O

Strong	O
CXCR4	B-Gene_or_gene_product
expression	O
was	O
significantly	O
associated	O
with	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastases	O
(	O
P	O
=	O
0	O
.	O
028	O
)	O
and	O
higher	O
stages	O
III	O
/	O
IV	O
(	O
P	O
=	O
0	O
.	O
047	O
)	O
,	O
and	O
further	O
tended	O
to	O
be	O
correlated	O
with	O
a	O
reduced	O
5	O
-	O
year	O
survival	O
rate	O
(	O
42	O
.	O
6	O
%	O
vs	O
.	O
53	O
.	O
9	O
%	O
;	O
P	O
=	O
0	O
.	O
1	O
)	O
.	O

In	O
conclusion	O
,	O
CXCR4	B-Gene_or_gene_product
expression	O
is	O
associated	O
with	O
gastric	B-Cell
cancer	I-Cell
cell	I-Cell
migration	O
in	O
vitro	O
,	O
and	O
strong	O
expression	O
of	O
CXCR4	B-Gene_or_gene_product
by	O
gastric	B-Cell
cancer	I-Cell
cells	I-Cell
is	O
significantly	O
associated	O
with	O
lymphatic	B-Organ
metastasis	O
in	O
patients	B-Organism
with	O
gastric	B-Cancer
cancer	I-Cancer
,	O
suggesting	O
that	O
CXCR4	B-Gene_or_gene_product
plays	O
an	O
important	O
role	O
during	O
gastric	B-Cancer
cancer	I-Cancer
progression	O
.	O

Effect	O
of	O
estrogen	B-Simple_chemical
and	O
progesterone	B-Simple_chemical
on	O
macrophage	B-Cell
activation	O
during	O
wound	B-Pathological_formation
healing	O
.	O

Age	O
-	O
related	O
impaired	O
wound	B-Pathological_formation
healing	O
leads	O
to	O
substantial	O
morbidity	O
and	O
mortality	O
along	O
with	O
a	O
large	O
financial	O
burden	O
to	O
health	O
services	O
.	O

There	O
is	O
accumulating	O
evidence	O
that	O
the	O
tissue	B-Tissue
damage	O
associated	O
with	O
chronic	O
wounds	B-Pathological_formation
is	O
initiated	O
and	O
propagated	O
by	O
an	O
inappropriately	O
excessive	O
inflammatory	O
response	O
.	O

Research	O
on	O
age	O
-	O
related	O
impaired	O
wound	B-Pathological_formation
healing	O
suggests	O
that	O
the	O
decline	O
in	O
sex	B-Simple_chemical
steroid	I-Simple_chemical
hormones	I-Simple_chemical
with	O
age	O
may	O
have	O
a	O
substantial	O
influence	O
on	O
the	O
inflammatory	O
response	O
in	O
vivo	O
.	O

Topical	O
and	O
systemic	O
estrogen	B-Simple_chemical
treatments	O
have	O
shown	O
an	O
increased	O
rate	O
of	O
healing	O
by	O
reducing	O
inflammation	O
,	O
however	O
the	O
underlying	O
mechanisms	O
are	O
little	O
understood	O
.	O

In	O
vitro	O
studies	O
also	O
suggest	O
progesterone	B-Simple_chemical
may	O
play	O
a	O
role	O
in	O
modulating	O
inflammation	O
.	O

Macrophages	B-Cell
are	O
essential	O
mediators	O
of	O
inflammation	O
and	O
wound	B-Pathological_formation
healing	O
.	O

Macrophages	B-Cell
can	O
be	O
activated	O
in	O
a	O
classical	O
or	O
alternative	O
manner	O
in	O
parallel	O
with	O
the	O
T	B-Cell
(	I-Cell
H	I-Cell
)	I-Cell
1	I-Cell
/	O
T	B-Cell
(	I-Cell
H	I-Cell
)	I-Cell
2	I-Cell
dichotomy	O
,	O
respectively	O
.	O

Using	O
a	O
murine	B-Organism
incisional	O
wound	B-Pathological_formation
healing	O
model	O
this	O
study	O
was	O
carried	O
out	O
to	O
investigate	O
the	O
roles	O
of	O
estrogen	B-Simple_chemical
and	O
progesterone	B-Simple_chemical
on	O
macrophage	B-Cell
activation	O
during	O
the	O
wound	B-Pathological_formation
healing	O
response	O
.	O

Our	O
findings	O
suggest	O
with	O
a	O
reduction	O
of	O
steroid	B-Simple_chemical
hormones	I-Simple_chemical
following	O
ovariectomy	O
,	O
alternatively	O
activated	O
macrophage	B-Cell
markers	O
(	O
Fizz1	B-Gene_or_gene_product
and	O
Ym1	B-Gene_or_gene_product
)	O
were	O
reduced	O
,	O
with	O
this	O
effect	O
being	O
reversed	O
with	O
the	O
administration	O
of	O
estrogen	B-Simple_chemical
or	O
progesterone	B-Simple_chemical
;	O
suggesting	O
that	O
with	O
the	O
reduction	O
of	O
steroid	B-Simple_chemical
hormones	I-Simple_chemical
macrophages	B-Cell
are	O
activated	O
in	O
a	O
classical	O
manner	O
,	O
promoting	O
inflammation	O
,	O
whereas	O
estrogen	B-Simple_chemical
or	O
progesterone	B-Simple_chemical
are	O
contributing	O
toward	O
macrophage	B-Cell
activation	O
in	O
an	O
alternative	O
manner	O
,	O
driving	O
wound	B-Pathological_formation
repair	O
,	O
angiogenesis	O
,	O
and	O
remodeling	O
.	O

Effect	O
of	O
platelet	B-Organism_substance
-	I-Organism_substance
rich	I-Organism_substance
plasma	I-Organism_substance
and	O
fibrin	B-Gene_or_gene_product
glue	O
on	O
healing	O
of	O
critical	O
-	O
size	O
calvarial	B-Organ
bone	I-Organ
defects	O
.	O

Despite	O
the	O
insufficient	O
number	O
of	O
experimental	O
studies	O
,	O
platelet	B-Organism_substance
-	I-Organism_substance
rich	I-Organism_substance
plasma	I-Organism_substance
(	O
PRP	B-Organism_substance
)	O
including	O
high	O
amounts	O
of	O
growth	O
factors	O
is	O
introduced	O
to	O
clinical	O
use	O
rapidly	O
.	O

The	O
aim	O
of	O
this	O
study	O
was	O
to	O
compare	O
the	O
effects	O
of	O
PRP	B-Organism_substance
and	O
platelet	B-Organism_substance
-	I-Organism_substance
poor	I-Organism_substance
plasma	I-Organism_substance
(	O
PPP	B-Organism_substance
)	O
on	O
healing	O
of	O
critical	O
-	O
size	O
bone	B-Organ
defects	O
.	O
Bilateral	O
full	O
-	O
thickness	O
,	O
critical	O
-	O
size	O
bone	B-Organ
defects	O
were	O
created	O
in	O
the	O
parietal	B-Organ
bones	I-Organ
of	O
32	O
rabbits	B-Organism
,	O
which	O
had	O
been	O
studied	O
in	O
4	O
groups	O
.	O

Saline	B-Simple_chemical
,	O
thrombin	B-Gene_or_gene_product
solution	O
,	O
PPP	B-Organism_substance
,	O
and	O
PRP	B-Organism_substance
were	O
applied	O
to	O
the	O
created	O
defects	O
before	O
closure	O
.	O

Radiologic	O
defect	O
area	O
measurement	O
results	O
at	O
0	O
,	O
4	O
,	O
and	O
16	O
weeks	O
were	O
compared	O
between	O
the	O
groups	O
.	O

In	O
addition	O
,	O
densities	O
of	O
the	O
newly	O
formed	O
bones	B-Organ
at	O
16th	O
week	O
were	O
studied	O
.	O

Histologic	O
parameters	O
(	O
primary	O
and	O
secondary	O
bone	B-Tissue
trabecula	I-Tissue
,	O
neovascularization	O
,	O
and	O
bone	B-Multi-tissue_structure
marrow	I-Multi-tissue_structure
and	O
connective	B-Tissue
tissue	I-Tissue
formation	O
)	O
were	O
compared	O
between	O
4	O
-	O
and	O
16	O
-	O
week	O
groups	O
.	O
More	O
rapid	O
decrease	O
in	O
defect	O
size	O
was	O
observed	O
in	O
groups	O
3	O
and	O
4	O
than	O
in	O
groups	O
1	O
and	O
2	O
,	O
both	O
in	O
the	O
4th	O
and	O
16th	O
weeks	O
.	O

Newly	O
formed	O
bone	B-Organ
densities	O
were	O
also	O
found	O
to	O
be	O
higher	O
in	O
these	O
2	O
groups	O
.	O

New	O
bone	B-Organ
formation	O
was	O
detected	O
to	O
be	O
more	O
rapid	O
considering	O
histologic	O
parameters	O
,	O
in	O
groups	O
3	O
and	O
4	O
at	O
4th	O
and	O
16th	O
weeks	O
.	O
Study	O
demonstrates	O
that	O
PRP	B-Organism_substance
and	O
PPP	B-Organism_substance
might	O
have	O
favorable	O
effects	O
on	O
bone	B-Organ
healing	O
.	O

Although	O
we	O
cannot	O
reveal	O
any	O
statistical	O
difference	O
between	O
these	O
2	O
substances	O
considering	O
osteoinductive	O
potential	O
,	O
PRP	B-Organism_substance
group	O
has	O
demonstrated	O
superior	O
results	O
compared	O
with	O
fibrin	B-Gene_or_gene_product
glue	O
group	O
.	O

Higher	O
platelet	B-Cell
concentrations	O
may	O
expose	O
beneficial	O
effects	O
of	O
PRP	B-Organism_substance
.	O

Host	O
-	O
derived	O
angiopoietin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
affects	O
early	O
stages	O
of	O
tumor	B-Cancer
development	O
and	O
vessel	B-Multi-tissue_structure
maturation	O
but	O
is	O
dispensable	O
for	O
later	O
stages	O
of	O
tumor	B-Cancer
growth	O
.	O

The	O
angiopoietin	B-Gene_or_gene_product
/	O
Tie2	B-Gene_or_gene_product
system	O
has	O
been	O
identified	O
as	O
the	O
second	O
vascular	B-Multi-tissue_structure
-	O
specific	O
receptor	O
tyrosine	O
kinase	O
system	O
controlling	O
vessel	B-Multi-tissue_structure
assembly	O
,	O
maturation	O
,	O
and	O
quiescence	O
.	O

Angiopoietin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
(	O
Ang	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
)	O
is	O
prominently	O
up	O
-	O
regulated	O
in	O
the	O
host	O
-	O
derived	O
vasculature	B-Multi-tissue_structure
of	O
most	O
tumors	B-Cancer
,	O
making	O
it	O
an	O
attractive	O
candidate	O
for	O
antiangiogenic	O
intervention	O
.	O

Yet	O
,	O
the	O
net	O
outcome	O
of	O
Ang	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
functions	O
on	O
tumor	B-Cancer
angiogenesis	O
is	O
believed	O
to	O
be	O
contextual	O
depending	O
on	O
the	O
local	O
cytokine	O
milieu	O
.	O

Correspondingly	O
,	O
Ang	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
manipulatory	O
therapies	O
have	O
been	O
shown	O
to	O
exert	O
protumorigenic	O
as	O
well	O
as	O
antitumorigenic	O
effects	O
.	O

To	O
clarify	O
the	O
role	O
of	O
Ang	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
for	O
angiogenesis	O
and	O
tumor	B-Cancer
growth	O
in	O
a	O
definite	O
genetic	O
experimental	O
setting	O
,	O
the	O
present	O
study	O
was	O
aimed	O
at	O
comparatively	O
studying	O
the	O
growth	O
of	O
different	O
tumors	B-Cancer
in	O
wild	O
-	O
type	O
and	O
Ang	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
-	O
deficient	O
mice	O
.	O

Lewis	B-Cancer
lung	I-Cancer
carcinomas	I-Cancer
,	O
MT	B-Cancer
-	I-Cancer
ret	I-Cancer
melanomas	I-Cancer
,	O
and	O
B16F10	B-Cancer
melanomas	I-Cancer
all	O
grew	O
slower	O
in	O
Ang	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
-	O
deficient	O
mice	O
.	O

Yet	O
,	O
tumor	B-Cancer
growth	O
in	O
wild	O
-	O
type	O
and	O
Ang	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
-	O
deficient	O
mice	O
dissociated	O
during	O
early	O
stages	O
of	O
tumor	B-Cancer
development	O
,	O
whereas	O
tumor	B-Cancer
growth	O
rates	O
during	O
later	O
stages	O
of	O
primary	O
tumor	B-Cancer
progression	O
were	O
similar	O
.	O

Analysis	O
of	O
the	O
intratumoral	B-Multi-tissue_structure
vascular	I-Multi-tissue_structure
architecture	I-Multi-tissue_structure
revealed	O
no	O
major	O
differences	O
in	O
microvessel	B-Tissue
density	O
and	O
perfusion	O
characteristics	O
.	O

However	O
,	O
diameters	O
of	O
intratumoral	B-Tissue
microvessels	I-Tissue
were	O
smaller	O
in	O
tumors	B-Cancer
grown	O
in	O
Ang	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
-	O
deficient	O
mice	O
,	O
and	O
the	O
vasculature	B-Multi-tissue_structure
had	O
an	O
altered	O
pattern	O
of	O
pericyte	B-Cell
recruitment	O
and	O
maturation	O
.	O

Ang	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
-	O
deficient	O
tumor	O
vessels	O
had	O
higher	O
pericyte	B-Cell
coverage	O
indices	O
.	O

Recruited	O
pericytes	B-Cell
were	O
desmin	B-Gene_or_gene_product
and	O
NG2	B-Gene_or_gene_product
positive	O
and	O
predominately	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
smooth	I-Gene_or_gene_product
muscle	I-Gene_or_gene_product
actin	I-Gene_or_gene_product
negative	O
,	O
indicative	O
of	O
a	O
more	O
mature	O
pericyte	B-Cell
phenotype	O
.	O

Collectively	O
,	O
the	O
experiments	O
define	O
the	O
role	O
of	O
Ang	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
during	O
tumor	B-Cancer
angiogenesis	O
and	O
establish	O
a	O
better	O
rationale	O
for	O
combination	O
therapies	O
involving	O
Ang	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
manipulatory	O
therapies	O
.	O

Liposomal	B-Cellular_component
honokiol	B-Simple_chemical
inhibits	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
D	I-Gene_or_gene_product
-	O
induced	O
lymphangiogenesis	O
and	O
metastasis	O
in	O
xenograft	B-Cancer
tumor	I-Cancer
model	O
.	O

Lymph	B-Multi-tissue_structure
nodes	I-Multi-tissue_structure
metastasis	O
of	O
tumor	B-Cancer
could	O
be	O
a	O
crucial	O
early	O
step	O
in	O
the	O
metastatic	O
process	O
.	O

Induction	O
of	O
tumor	B-Cancer
lymphangiogenesis	O
by	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
D	I-Gene_or_gene_product
may	O
play	O
an	O
important	O
role	O
in	O
promoting	O
tumor	B-Cancer
metastasis	O
to	O
regional	O
lymph	B-Multi-tissue_structure
nodes	I-Multi-tissue_structure
and	O
these	O
processes	O
can	O
be	O
inhibited	O
by	O
inactivation	O
of	O
the	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
signaling	O
pathway	O
.	O

Honokiol	B-Simple_chemical
has	O
been	O
reported	O
to	O
possess	O
potent	O
antiangiogenesis	O
and	O
antitumor	B-Cancer
properties	O
in	O
several	O
cell	B-Cell
lines	I-Cell
and	O
xenograft	B-Cancer
tumor	I-Cancer
models	O
.	O

However	O
,	O
its	O
role	O
in	O
tumor	B-Cancer
-	O
associated	O
lymphangiogenesis	O
and	O
lymphatic	B-Organ
metastasis	O
remains	O
unclear	O
.	O

Here	O
,	O
we	O
established	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
models	O
by	O
injecting	O
overexpressing	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
D	I-Gene_or_gene_product
Lewis	B-Cell
lung	I-Cell
carcinoma	I-Cell
cells	I-Cell
into	O
C57BL	B-Organism
/	I-Organism
6	I-Organism
mice	I-Organism
to	O
explore	O
the	O
effect	O
of	O
honokiol	B-Simple_chemical
on	O
tumor	B-Cancer
-	O
associated	O
lymphangiogenesis	O
and	O
related	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
.	O

The	O
underlying	O
mechanisms	O
were	O
systematically	O
investigated	O
in	O
vitro	O
and	O
in	O
vivo	O
.	O

In	O
in	O
vivo	O
study	O
,	O
liposomal	B-Cellular_component
honokiol	B-Simple_chemical
significantly	O
inhibited	O
the	O
tumor	B-Cancer
-	O
associated	O
lymphangiogenesis	O
and	O
metastasis	O
in	O
Lewis	B-Cancer
lung	I-Cancer
carcinoma	I-Cancer
model	O
.	O

A	O
remarkable	O
delay	O
of	O
tumor	B-Cancer
growth	O
and	O
prolonged	O
life	O
span	O
were	O
also	O
observed	O
.	O

In	O
in	O
vitro	O
study	O
,	O
honokiol	B-Simple_chemical
inhibited	O
VEGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
D	I-Gene_or_gene_product
-	O
induced	O
survival	O
,	O
proliferation	O
and	O
tube	B-Tissue
-	I-Tissue
formation	O
of	O
both	O
human	B-Cell
umbilical	I-Cell
vein	I-Cell
endothelial	I-Cell
cells	I-Cell
(	O
HUVECs	B-Cell
)	O
and	O
lymphatic	B-Cell
vascular	I-Cell
endothelial	I-Cell
cells	I-Cell
(	O
HLECs	B-Cell
)	O
.	O

Western	O
blotting	O
analysis	O
showed	O
that	O
liposomal	B-Cellular_component
honokiol	B-Simple_chemical
-	O
inhibited	O
Akt	B-Gene_or_gene_product
and	O
MAPK	B-Gene_or_gene_product
phosphorylation	O
in	O
2	O
endothelial	B-Cell
cells	I-Cell
,	O
and	O
downregulated	O
expressions	O
of	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
of	O
human	B-Organism
vascular	B-Cell
endothelial	I-Cell
cells	I-Cell
and	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
of	O
lymphatic	B-Cell
endothelial	I-Cell
cells	I-Cell
.	O

Thus	O
,	O
we	O
identified	O
for	O
the	O
first	O
time	O
that	O
honokiol	B-Simple_chemical
provided	O
therapeutic	O
benefit	O
not	O
only	O
by	O
direct	O
effects	O
on	O
tumor	B-Cell
cells	I-Cell
and	O
antiangiogenesis	O
but	O
also	O
by	O
inhibiting	O
lymphangiogenesis	O
and	O
metastasis	O
via	O
the	O
VEGFR	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
pathway	O
.	O

The	O
present	O
findings	O
may	O
be	O
of	O
importance	O
to	O
investigate	O
the	O
molecular	O
mechanisms	O
underlying	O
the	O
spread	O
of	O
cancer	B-Cancer
via	O
the	O
lymphatics	B-Multi-tissue_structure
and	O
explore	O
the	O
therapeutical	O
strategy	O
of	O
honokiol	B-Simple_chemical
on	O
antilymphangiogenesis	O
and	O
antimetastasis	O
.	O

Short	B-Gene_or_gene_product
pigment	I-Gene_or_gene_product
epithelial	I-Gene_or_gene_product
-	I-Gene_or_gene_product
derived	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
derived	I-Gene_or_gene_product
peptide	I-Gene_or_gene_product
inhibits	O
angiogenesis	O
and	O
tumor	B-Cancer
growth	O
.	O

PURPOSE	O
:	O
Pigment	B-Gene_or_gene_product
epithelial	I-Gene_or_gene_product
-	I-Gene_or_gene_product
derived	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
PEDF	B-Gene_or_gene_product
)	O
is	O
a	O
potent	O
angiogenesis	O
inhibitor	O
with	O
multiple	O
other	O
functions	O
,	O
some	O
of	O
which	O
enhance	O
tumor	B-Cancer
growth	O
.	O

Our	O
previous	O
studies	O
mapped	O
PEDF	B-Gene_or_gene_product
antiangiogenic	O
and	O
prosurvival	O
activities	O
to	O
distinct	O
epitopes	O
.	O

This	O
study	O
was	O
aimed	O
to	O
determine	O
the	O
minimal	O
fragment	O
of	O
PEDF	B-Gene_or_gene_product
,	O
which	O
maintains	O
antiangiogenic	O
and	O
antitumor	B-Cancer
efficacy	O
.	O

EXPERIMENTAL	O
DESIGN	O
:	O
We	O
analyzed	O
antigenicity	O
,	O
hydrophilicity	O
,	O
and	O
charge	O
distribution	O
of	O
the	O
angioinhibitory	O
epitope	O
(	O
the	O
34	O
-	O
mer	O
)	O
and	O
designed	O
three	O
peptides	O
covering	O
its	O
COOH	O
terminus	O
,	O
P14	B-Gene_or_gene_product
,	O
P18	B-Gene_or_gene_product
,	O
and	O
P23	B-Gene_or_gene_product
.	O

We	O
analyzed	O
their	O
ability	O
to	O
block	O
endothelial	B-Cell
cell	I-Cell
chemotaxis	O
and	O
induce	O
apoptosis	O
in	O
vitro	O
and	O
their	O
antiangiogenic	O
activity	O
in	O
vivo	O
.	O

The	O
selected	O
peptide	O
was	O
tested	O
for	O
the	O
antitumor	B-Cancer
activity	O
against	O
mildly	O
aggressive	O
xenografted	O
prostate	B-Cancer
carcinoma	I-Cancer
and	O
highly	O
aggressive	O
renal	B-Cancer
cell	I-Cancer
carcinoma	I-Cancer
.	O

To	O
verify	O
that	O
P18	B-Gene_or_gene_product
acts	O
in	O
the	O
same	O
manner	O
as	O
PEDF	B-Gene_or_gene_product
,	O
we	O
used	O
immunohistochemistry	O
to	O
measure	O
PEDF	B-Gene_or_gene_product
targets	I-Gene_or_gene_product
,	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
and	O
CD95	B-Gene_or_gene_product
ligand	I-Gene_or_gene_product
expression	O
in	O
P18	B-Gene_or_gene_product
-	O
treated	O
vasculature	B-Multi-tissue_structure
.	O

RESULTS	O
:	O
P14	B-Gene_or_gene_product
and	O
P18	B-Gene_or_gene_product
blocked	O
endothelial	B-Cell
cell	I-Cell
chemotaxis	O
;	O
P18	B-Gene_or_gene_product
and	O
P23	B-Gene_or_gene_product
induced	O
apoptosis	O
.	O

P18	B-Gene_or_gene_product
showed	O
the	O
highest	O
IC50	O
and	O
blocked	O
angiogenesis	O
in	O
vivo	O
:	O
P23	B-Gene_or_gene_product
was	O
inactive	O
and	O
P14	B-Gene_or_gene_product
was	O
proangiogenic	O
.	O

P18	B-Gene_or_gene_product
increased	O
the	O
production	O
of	O
CD95	B-Gene_or_gene_product
ligand	I-Gene_or_gene_product
and	O
reduced	O
the	O
expression	O
of	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
receptor	I-Gene_or_gene_product
2	I-Gene_or_gene_product
by	O
the	O
endothelial	B-Cell
cells	I-Cell
in	O
vivo	O
.	O

In	O
tumor	B-Cancer
studies	O
,	O
P18	B-Gene_or_gene_product
was	O
more	O
effective	O
in	O
blocking	O
the	O
angiogenesis	O
and	O
growth	O
of	O
the	O
prostate	B-Cancer
cancer	I-Cancer
than	O
parental	B-Gene_or_gene_product
34	I-Gene_or_gene_product
-	I-Gene_or_gene_product
mer	I-Gene_or_gene_product
;	O
in	O
the	O
renal	B-Cancer
cell	I-Cancer
carcinoma	I-Cancer
,	O
P18	B-Gene_or_gene_product
strongly	O
decreased	O
angiogenesis	O
and	O
halted	O
the	O
progression	O
of	O
established	O
tumors	B-Cancer
.	O

CONCLUSIONS	O
:	O
P18	B-Gene_or_gene_product
is	O
a	O
novel	O
and	O
potent	O
antiangiogenic	O
biotherapeutic	O
agent	O
that	O
has	O
potential	O
to	O
be	O
developed	O
for	O
the	O
treatment	O
of	O
prostate	B-Cancer
and	O
renal	B-Cancer
cancer	I-Cancer
.	O

Distinct	O
role	O
of	O
PLCbeta3	B-Gene_or_gene_product
in	O
VEGF	B-Gene_or_gene_product
-	O
mediated	O
directional	O
migration	O
and	O
vascular	B-Multi-tissue_structure
sprouting	O
.	O

Endothelial	B-Cell
cell	I-Cell
proliferation	O
and	O
migration	O
is	O
essential	O
to	O
angiogenesis	O
.	O

Typically	O
,	O
proliferation	O
and	O
chemotaxis	O
of	O
endothelial	B-Cell
cells	I-Cell
is	O
driven	O
by	O
growth	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
and	O
basic	B-Gene_or_gene_product
fibroblast	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
bFGF	B-Gene_or_gene_product
)	O
.	O

VEGF	B-Gene_or_gene_product
activates	O
phospholipases	B-Gene_or_gene_product
(	O
PLCs	B-Gene_or_gene_product
)	O
-	O
specifically	O
PLCgamma1	B-Gene_or_gene_product
-	O
that	O
are	O
important	O
for	O
tubulogenesis	O
,	O
differentiation	O
and	O
DNA	B-Cellular_component
synthesis	O
.	O

However	O
,	O
we	O
show	O
here	O
that	O
VEGF	B-Gene_or_gene_product
,	O
specifically	O
through	O
VEGFR2	B-Gene_or_gene_product
,	O
induces	O
phosphorylation	O
of	O
two	O
serine	O
residues	O
on	O
PLCbeta3	B-Gene_or_gene_product
,	O
and	O
this	O
was	O
confirmed	O
in	O
an	O
ex	O
vivo	O
embryoid	B-Developing_anatomical_structure
body	I-Developing_anatomical_structure
model	O
.	O

Knockdown	O
of	O
PLCbeta3	B-Gene_or_gene_product
in	O
HUVEC	B-Cell
cells	I-Cell
affects	O
IP3	B-Gene_or_gene_product
production	O
,	O
actin	B-Gene_or_gene_product
reorganization	O
,	O
migration	O
and	O
proliferation	O
;	O
whereas	O
migration	O
is	O
inhibited	O
,	O
proliferation	O
is	O
enhanced	O
.	O

Our	O
data	O
suggest	O
that	O
enhanced	O
proliferation	O
is	O
precipitated	O
by	O
an	O
accelerated	O
cell	B-Cell
cycle	O
,	O
and	O
decreased	O
migration	O
by	O
an	O
inability	O
to	O
activate	O
CDC42	B-Gene_or_gene_product
.	O

Given	O
that	O
PLCbeta3	B-Gene_or_gene_product
is	O
typically	O
known	O
as	O
an	O
effector	O
of	O
heterotrimeric	B-Gene_or_gene_product
G	I-Gene_or_gene_product
-	I-Gene_or_gene_product
proteins	I-Gene_or_gene_product
,	O
our	O
data	O
demonstrate	O
a	O
unique	O
crosstalk	O
between	O
the	O
G	B-Gene_or_gene_product
-	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
and	O
receptor	O
tyrosine	O
kinase	O
(	O
RTK	O
)	O
axes	O
and	O
reveal	O
a	O
novel	O
molecular	O
mechanism	O
of	O
VEGF	B-Gene_or_gene_product
signaling	O
and	O
,	O
thus	O
,	O
angiogenesis	O
.	O

Antivascular	B-Multi-tissue_structure
actions	O
of	O
microtubule	B-Simple_chemical
-	I-Simple_chemical
binding	I-Simple_chemical
drugs	I-Simple_chemical
.	O

Microtubule	B-Simple_chemical
-	I-Simple_chemical
binding	I-Simple_chemical
drugs	I-Simple_chemical
(	O
MBD	B-Simple_chemical
)	O
are	O
widely	O
used	O
in	O
cancer	B-Cancer
chemotherapy	O
and	O
also	O
have	O
clinically	O
relevant	O
antiangiogenic	O
and	O
vascular	B-Multi-tissue_structure
-	O
disrupting	O
properties	O
.	O

These	O
antivascular	B-Multi-tissue_structure
actions	O
are	O
due	O
in	O
part	O
to	O
direct	O
effects	O
on	O
endothelial	B-Cell
cells	I-Cell
,	O
and	O
all	O
MBDs	B-Simple_chemical
(	O
both	O
microtubule	B-Cellular_component
-	O
stabilizing	O
and	O
microtubule	B-Cellular_component
-	O
destabilizing	O
)	O
inhibit	O
endothelial	B-Cell
cell	I-Cell
proliferation	O
,	O
migration	O
,	O
and	O
tube	B-Tissue
formation	O
in	O
vitro	O
,	O
actions	O
that	O
are	O
thought	O
to	O
correspond	O
to	O
therapeutic	O
antiangiogenic	O
actions	O
.	O

In	O
addition	O
,	O
the	O
microtubule	B-Simple_chemical
-	I-Simple_chemical
destabilizing	I-Simple_chemical
agents	I-Simple_chemical
cause	O
prominent	O
changes	O
in	O
endothelial	B-Cell
cell	I-Cell
morphology	O
,	O
an	O
action	O
associated	O
with	O
rapid	O
vascular	B-Multi-tissue_structure
collapse	O
in	O
vivo	O
.	O

The	O
effects	O
on	O
endothelial	B-Cell
cells	I-Cell
occur	O
in	O
vitro	O
at	O
low	O
drug	O
concentrations	O
,	O
which	O
do	O
not	O
affect	O
microtubule	B-Cellular_component
gross	O
morphology	O
,	O
do	O
not	O
cause	O
microtubule	B-Cellular_component
bundling	O
or	O
microtubule	B-Cellular_component
loss	O
and	O
do	O
not	O
induce	O
cell	B-Cell
cycle	O
arrest	O
,	O
apoptosis	O
,	O
or	O
cell	B-Cell
death	O
.	O

Rather	O
,	O
it	O
has	O
been	O
hypothesized	O
that	O
,	O
at	O
low	O
concentrations	O
,	O
MBDs	B-Simple_chemical
produce	O
more	O
subtle	O
effects	O
on	O
microtubule	B-Cellular_component
dynamics	O
,	O
block	O
critical	O
cell	B-Cell
signaling	O
pathways	O
,	O
and	O
prevent	O
the	O
microtubules	B-Cellular_component
from	O
properly	O
interacting	O
with	O
transient	O
subcellular	B-Cellular_component
assemblies	I-Cellular_component
(	O
focal	B-Cellular_component
adhesions	I-Cellular_component
and	O
adherens	B-Cellular_component
junctions	I-Cellular_component
)	O
whose	O
subsequent	O
stabilization	O
and	O
/	O
or	O
maturation	O
are	O
required	O
for	O
cell	B-Cell
motility	O
and	O
cell	B-Cell
-	O
cell	B-Cell
interactions	O
.	O

This	O
review	O
will	O
focus	O
on	O
recent	O
studies	O
to	O
define	O
the	O
molecular	O
mechanisms	O
for	O
the	O
antivascular	B-Multi-tissue_structure
actions	O
of	O
the	O
MBDs	B-Simple_chemical
,	O
information	O
that	O
could	O
be	O
useful	O
in	O
the	O
identification	O
or	O
design	O
of	O
agents	O
whose	O
actions	O
more	O
selectively	O
target	O
the	O
tumor	B-Multi-tissue_structure
vasculature	I-Multi-tissue_structure
.	O

Neurovascular	B-Multi-tissue_structure
effects	O
of	O
CD47	B-Gene_or_gene_product
signaling	O
:	O
promotion	O
of	O
cell	B-Cell
death	O
,	O
inflammation	O
,	O
and	O
suppression	O
of	O
angiogenesis	O
in	O
brain	B-Cell
endothelial	I-Cell
cells	I-Cell
in	O
vitro	O
.	O

The	O
concept	O
of	O
the	O
neurovascular	B-Multi-tissue_structure
unit	O
emphasizes	O
that	O
common	O
signals	O
and	O
substrates	O
underlie	O
the	O
physiology	O
and	O
pathophysiology	O
of	O
neuronal	B-Multi-tissue_structure
and	O
endothelial	B-Multi-tissue_structure
compartments	I-Multi-tissue_structure
in	O
brain	B-Organ
.	O

Recent	O
data	O
suggest	O
that	O
activation	O
of	O
the	O
integrin	B-Gene_or_gene_product
-	O
associated	O
protein	O
CD47	B-Gene_or_gene_product
promotes	O
neuronal	B-Cell
cell	I-Cell
death	O
.	O

Is	O
it	O
possible	O
that	O
CD47	B-Gene_or_gene_product
may	O
also	O
negatively	O
affect	O
cerebral	B-Cell
endothelial	I-Cell
cells	I-Cell
?	O

Exposure	O
of	O
wild	O
-	O
type	O
primary	O
mouse	B-Organism
cerebral	B-Cell
endothelial	I-Cell
cells	I-Cell
to	O
the	O
CD47	B-Gene_or_gene_product
ligand	O
thrombospondin	B-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
induced	O
an	O
increasing	O
amount	O
of	O
cell	B-Cell
death	O
,	O
whereas	O
cytotoxicity	O
was	O
significantly	O
decreased	O
in	O
cerebral	B-Cell
endothelial	I-Cell
cells	I-Cell
derived	O
from	O
CD47	B-Gene_or_gene_product
knockout	O
mice	O
.	O

The	O
specific	O
CD47	B-Gene_or_gene_product
-	O
activating	O
peptide	O
,	O
4N1K	B-Gene_or_gene_product
,	O
similarly	O
induced	O
cell	B-Cell
death	O
in	O
human	B-Organism
brain	B-Cell
microvascular	I-Cell
endothelial	I-Cell
cells	I-Cell
.	O

Promotion	O
of	O
inflammation	O
was	O
also	O
involved	O
because	O
lower	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
was	O
able	O
to	O
up	O
-	O
regulate	O
the	O
adhesion	O
molecules	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
and	O
vascular	B-Gene_or_gene_product
cell	I-Gene_or_gene_product
adhesion	I-Gene_or_gene_product
molecule	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

Finally	O
,	O
CD47	B-Gene_or_gene_product
signaling	O
may	O
suppress	O
angiogenesis	O
because	O
4N1K	B-Gene_or_gene_product
significantly	O
inhibited	O
endothelial	B-Cell
cell	I-Cell
migration	O
and	O
tube	B-Tissue
formation	O
in	O
vitro	O
.	O

We	O
conclude	O
that	O
CD47	B-Gene_or_gene_product
signaling	O
can	O
negatively	O
affect	O
the	O
viability	O
and	O
function	O
of	O
cerebral	B-Cell
endothelial	I-Cell
cells	I-Cell
,	O
further	O
supporting	O
the	O
notion	O
that	O
CD47	B-Gene_or_gene_product
may	O
be	O
a	O
potential	O
neurovascular	B-Multi-tissue_structure
target	O
for	O
stroke	O
and	O
brain	B-Organ
injury	O
.	O

Cisplatin	B-Simple_chemical
reduces	O
endothelial	B-Cell
cell	I-Cell
migration	O
via	O
regulation	O
of	O
type	B-Gene_or_gene_product
2	I-Gene_or_gene_product
-	I-Gene_or_gene_product
matrix	I-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
activity	O
.	O

AIMS	O
:	O
In	O
this	O
study	O
we	O
investigated	O
the	O
effect	O
of	O
cisplatin	B-Simple_chemical
on	O
endothelial	B-Cell
cell	I-Cell
migration	O
,	O
an	O
essential	O
process	O
for	O
vascular	B-Multi-tissue_structure
remodeling	O
and	O
regeneration	O
in	O
several	O
physiological	O
and	O
pathological	O
situations	O
.	O

MATERIAL	O
AND	O
METHODS	O
:	O
Human	B-Cell
umbilical	I-Cell
vein	I-Cell
endothelial	I-Cell
cells	I-Cell
(	O
HUVEC	B-Cell
)	O
were	O
treated	O
with	O
cisplatin	B-Simple_chemical
and	O
endothelial	B-Cell
cell	I-Cell
migration	O
analyzed	O
by	O
fluorescence	O
and	O
scratch	B-Pathological_formation
-	I-Pathological_formation
wound	I-Pathological_formation
migration	O
assay	O
.	O

MMP2	B-Gene_or_gene_product
and	O
MMP9	B-Gene_or_gene_product
activity	O
were	O
determined	O
by	O
zymographic	O
assay	O
,	O
and	O
MAPK	B-Gene_or_gene_product
activation	O
by	O
Western	O
blotting	O
analysis	O
.	O

RESULTS	O
:	O
We	O
demonstrated	O
that	O
cisplatin	B-Simple_chemical
provoked	O
a	O
time	O
-	O
and	O
dose	O
-	O
dependent	O
decrease	O
of	O
HUVEC	B-Cell
migration	O
;	O
this	O
effect	O
was	O
clearly	O
independent	O
from	O
its	O
well	O
known	O
cytotoxic	O
activity	O
.	O

In	O
addition	O
,	O
cisplatin	B-Simple_chemical
markedly	O
reduced	O
MMP2	B-Gene_or_gene_product
activity	O
in	O
both	O
conditioned	O
media	O
and	O
cell	B-Organism_substance
lysates	I-Organism_substance
,	O
increased	O
p38	B-Gene_or_gene_product
MAPK	I-Gene_or_gene_product
and	O
JNK	B-Gene_or_gene_product
phosphorylation	O
,	O
but	O
did	O
not	O
affect	O
ERK	B-Gene_or_gene_product
phosphorylation	O
.	O

Endothelial	B-Cell
cell	I-Cell
migration	O
was	O
attenuated	O
by	O
treatment	O
of	O
cells	B-Cell
with	O
GM6001	B-Simple_chemical
,	O
a	O
non	O
-	O
specific	O
inhibitor	O
of	O
MMPs	B-Gene_or_gene_product
,	O
or	O
by	O
a	O
selective	O
anti	O
-	O
MMP2	B-Gene_or_gene_product
antibody	O
.	O

However	O
,	O
treatment	O
of	O
cells	B-Cell
with	O
SB202190	B-Simple_chemical
or	O
SP600125	B-Simple_chemical
,	O
inhibitors	O
of	O
p38	B-Gene_or_gene_product
MAPK	I-Gene_or_gene_product
and	O
JNK	B-Gene_or_gene_product
respectively	O
,	O
did	O
not	O
affect	O
HUVEC	B-Cell
migration	O
.	O

CONCLUSION	O
:	O
These	O
results	O
suggested	O
that	O
cisplatin	B-Simple_chemical
induced	O
a	O
reduction	O
of	O
endothelial	B-Cell
cell	I-Cell
migration	O
through	O
an	O
inhibition	O
of	O
MMP2	B-Gene_or_gene_product
activity	O
by	O
downstream	O
signal	O
transduction	O
pathways	O
independent	O
of	O
JNK	B-Gene_or_gene_product
and	O
p38	B-Gene_or_gene_product
MAPK	I-Gene_or_gene_product
activation	O
.	O

The	O
modulation	O
of	O
platelet	B-Cell
and	O
endothelial	B-Cell
cell	I-Cell
adhesion	O
to	O
vascular	B-Multi-tissue_structure
graft	I-Multi-tissue_structure
materials	O
by	O
perlecan	B-Gene_or_gene_product
.	O

Controlled	O
neo	O
-	O
endothelialisation	O
is	O
critical	O
to	O
the	O
patency	O
of	O
small	O
diameter	O
vascular	B-Multi-tissue_structure
grafts	I-Multi-tissue_structure
.	O

Endothelialisation	O
and	O
platelet	B-Cell
adhesion	O
to	O
purified	O
endothelial	B-Cell
cell	I-Cell
-	O
derived	O
perlecan	B-Gene_or_gene_product
,	O
the	O
major	O
heparan	B-Gene_or_gene_product
sulfate	I-Gene_or_gene_product
(	I-Gene_or_gene_product
HS	I-Gene_or_gene_product
)	I-Gene_or_gene_product
proteoglycan	I-Gene_or_gene_product
in	O
basement	B-Cellular_component
membranes	I-Cellular_component
,	O
were	O
investigated	O
using	O
in	O
vivo	O
and	O
in	O
vitro	O
assays	O
.	O

Expanded	O
polytetrafluoroethylene	B-Simple_chemical
(	O
ePTFE	B-Simple_chemical
)	O
vascular	B-Multi-tissue_structure
grafts	I-Multi-tissue_structure
were	O
coated	O
with	O
perlecan	B-Gene_or_gene_product
and	O
tested	O
in	O
an	O
ovine	B-Organism
carotid	B-Multi-tissue_structure
interposition	O
model	O
for	O
a	O
period	O
of	O
6	O
weeks	O
and	O
assessed	O
using	O
light	O
and	O
scanning	O
microscopy	O
.	O

Enhanced	O
endothelial	B-Cell
cell	I-Cell
growth	O
and	O
reduced	O
platelet	B-Cell
adhesion	O
were	O
observed	O
on	O
the	O
perlecan	B-Gene_or_gene_product
coated	O
grafts	B-Multi-tissue_structure
when	O
compared	O
to	O
uncoated	O
controls	O
implanted	O
in	O
the	O
same	O
sheep	B-Organism
(	O
n	O
=	O
5	O
)	O
.	O

Perlecan	B-Gene_or_gene_product
was	O
also	O
found	O
to	O
stimulate	O
endothelial	B-Cell
cell	I-Cell
proliferation	O
in	O
vitro	O
over	O
a	O
period	O
of	O
6	O
days	O
in	O
the	O
presence	O
of	O
plasma	B-Organism_substance
proteins	O
and	O
fibroblastic	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
2	I-Gene_or_gene_product
(	O
FGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
)	O
,	O
however	O
in	O
the	O
absence	O
of	O
FGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
endothelial	B-Cell
cell	I-Cell
growth	O
could	O
not	O
be	O
maintained	O
during	O
this	O
period	O
.	O

Perlecan	B-Gene_or_gene_product
was	O
found	O
to	O
be	O
anti	O
-	O
adhesive	O
for	O
platelets	B-Cell
,	O
however	O
after	O
removal	O
of	O
the	O
HS	O
chains	O
attached	O
to	O
perlecan	B-Gene_or_gene_product
,	O
platelet	B-Cell
adhesion	O
and	O
aggregation	O
were	O
supported	O
.	O

These	O
results	O
suggest	O
a	O
role	O
for	O
HS	O
chains	O
of	O
perlecan	B-Gene_or_gene_product
in	O
improving	O
graft	B-Multi-tissue_structure
patency	O
by	O
selectively	O
promoting	O
endothelial	B-Cell
cell	I-Cell
proliferation	O
while	O
modulating	O
platelet	B-Cell
adhesion	O
.	O

Tumour	B-Cancer
angiogenesis	O
:	O
its	O
mechanism	O
and	O
therapeutic	O
implications	O
in	O
malignant	B-Cancer
gliomas	I-Cancer
.	O

Angiogenesis	O
is	O
a	O
key	O
event	O
in	O
the	O
progression	O
of	O
malignant	B-Cancer
gliomas	I-Cancer
.	O

The	O
presence	O
of	O
microvascular	B-Tissue
proliferation	O
leads	O
to	O
the	O
histological	O
diagnosis	O
of	O
glioblastoma	B-Cancer
multiforme	I-Cancer
.	O

Tumour	B-Cancer
angiogenesis	O
involves	O
multiple	O
cellular	B-Cell
processes	O
including	O
endothelial	B-Cell
cell	I-Cell
proliferation	O
,	O
migration	O
,	O
reorganisation	O
of	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
and	O
tube	B-Tissue
formation	O
.	O

These	O
processes	O
are	O
regulated	O
by	O
numerous	O
pro	O
-	O
angiogenic	O
and	O
anti	O
-	O
angiogenic	O
growth	O
factors	O
.	O

Angiogenesis	O
inhibitors	O
have	O
been	O
developed	O
to	O
interrupt	O
the	O
angiogenic	O
process	O
at	O
the	O
growth	O
factor	O
,	O
receptor	O
tyrosine	O
kinase	O
and	O
intracellular	B-Immaterial_anatomical_entity
kinase	O
levels	O
.	O

Other	O
anti	O
-	O
angiogenic	O
therapies	O
alter	O
the	O
immune	O
response	O
and	O
endogeneous	O
angiogenesis	O
inhibitor	O
levels	O
.	O

Most	O
anti	O
-	O
angiogenic	O
therapies	O
for	O
malignant	B-Cancer
gliomas	I-Cancer
are	O
in	O
Phase	O
I	O
/	O
II	O
trials	O
and	O
only	O
modest	O
efficacies	O
are	O
reported	O
for	O
monotherapies	O
.	O

The	O
greatest	O
potential	O
for	O
angiogenesis	O
inhibitors	O
may	O
lie	O
in	O
their	O
ability	O
to	O
combine	O
safely	O
with	O
chemotherapy	O
and	O
radiotherapy	O
.	O

Regulation	O
of	O
thrombospondin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
by	O
natural	O
and	O
synthetic	O
progestins	B-Simple_chemical
in	O
human	B-Organism
breast	B-Cell
cancer	I-Cell
cells	I-Cell
.	O

Our	O
recent	O
studies	O
show	O
that	O
progestins	B-Simple_chemical
induce	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
breast	B-Cell
cancer	I-Cell
cells	I-Cell
that	O
express	O
mutant	O
p53	B-Gene_or_gene_product
protein	O
.	O

Here	O
,	O
we	O
show	O
that	O
natural	O
and	O
synthetic	O
progestins	B-Simple_chemical
also	O
induce	O
thrombospondin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
mRNA	O
and	O
protein	O
in	O
T47	B-Cell
-	I-Cell
D	I-Cell
and	O
BT	B-Cell
-	I-Cell
474	I-Cell
breast	I-Cell
cancer	I-Cell
cells	I-Cell
.	O

Antiprogestin	B-Simple_chemical
RU	I-Simple_chemical
-	I-Simple_chemical
486	I-Simple_chemical
inhibits	O
the	O
induction	O
of	O
VEGF	B-Gene_or_gene_product
and	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
by	O
progestins	B-Simple_chemical
,	O
suggesting	O
that	O
this	O
effect	O
of	O
progestin	B-Simple_chemical
is	O
mediated	O
by	O
the	O
progesterone	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
(	O
PR	B-Gene_or_gene_product
)	O
.	O

Actinomycin	B-Simple_chemical
-	I-Simple_chemical
D	I-Simple_chemical
,	O
but	O
not	O
puromycin	B-Simple_chemical
,	O
also	O
blocks	O
progestin	B-Simple_chemical
-	O
dependent	O
induction	O
of	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

A	O
putative	O
progestin	B-Simple_chemical
-	O
response	O
element	O
was	O
identified	O
in	O
the	O
human	B-Organism
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
promoter	O
,	O
which	O
is	O
consistent	O
with	O
the	O
hypothesis	O
that	O
a	O
progestin	B-Simple_chemical
-	O
PR	B-Gene_or_gene_product
complex	O
might	O
directly	O
regulate	O
transcription	O
of	O
the	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
gene	O
in	O
human	B-Organism
cells	B-Cell
.	O

Conditioned	O
medium	O
from	O
progestin	B-Simple_chemical
-	O
treated	O
breast	B-Cell
cancer	I-Cell
cells	I-Cell
stimulates	O
endothelial	B-Cell
cell	I-Cell
proliferation	O
in	O
the	O
absence	O
though	O
not	O
in	O
the	O
presence	O
of	O
antibody	B-Gene_or_gene_product
to	I-Gene_or_gene_product
TSP	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
indicating	O
that	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
secreted	O
by	O
breast	B-Cell
cancer	I-Cell
cells	I-Cell
could	O
be	O
pro	O
-	O
angiogenic	O
.	O

Since	O
tumor	B-Cell
cell	I-Cell
-	O
derived	O
TSP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
has	O
the	O
potential	O
to	O
promote	O
angiogenesis	O
in	O
the	O
tumor	B-Cancer
microenvironment	O
,	O
it	O
could	O
be	O
a	O
potential	O
target	O
for	O
breast	B-Cancer
cancer	I-Cancer
therapy	O
.	O

WNT	B-Gene_or_gene_product
/	O
TCF	B-Gene_or_gene_product
signaling	O
through	O
LEF1	B-Gene_or_gene_product
and	O
HOXB9	B-Gene_or_gene_product
mediates	O
lung	B-Cancer
adenocarcinoma	I-Cancer
metastasis	O
.	O

Metastasis	O
from	O
lung	B-Cancer
adenocarcinoma	I-Cancer
can	O
occur	O
swiftly	O
to	O
multiple	O
organs	B-Organ
within	O
months	O
of	O
diagnosis	O
.	O

The	O
mechanisms	O
that	O
confer	O
this	O
rapid	O
metastatic	O
capacity	O
to	O
lung	B-Cancer
tumors	I-Cancer
are	O
unknown	O
.	O

Activation	O
of	O
the	O
canonical	O
WNT	B-Gene_or_gene_product
/	O
TCF	B-Gene_or_gene_product
pathway	O
is	O
identified	O
here	O
as	O
a	O
determinant	O
of	O
metastasis	O
to	O
brain	B-Organ
and	O
bone	B-Tissue
during	O
lung	B-Cancer
adenocarcinoma	I-Cancer
progression	O
.	O

Gene	O
expression	O
signatures	O
denoting	O
WNT	B-Gene_or_gene_product
/	O
TCF	B-Gene_or_gene_product
activation	O
are	O
associated	O
with	O
relapse	O
to	O
multiple	O
organs	B-Organ
in	O
primary	O
lung	B-Cancer
adenocarcinoma	I-Cancer
.	O

Metastatic	O
subpopulations	O
isolated	O
from	O
independent	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
-	O
derived	O
lung	B-Cell
adenocarcinoma	I-Cell
cell	I-Cell
lines	I-Cell
harbor	O
a	O
hyperactive	O
WNT	B-Gene_or_gene_product
/	O
TCF	B-Gene_or_gene_product
pathway	O
.	O

Reduction	O
of	O
TCF	B-Gene_or_gene_product
activity	O
in	O
these	O
cells	B-Cell
attenuates	O
their	O
ability	O
to	O
form	O
brain	B-Organ
and	O
bone	B-Organ
metastases	O
in	O
mice	B-Organism
,	O
independently	O
of	O
effects	O
on	O
tumor	B-Cancer
growth	O
in	O
the	O
lungs	B-Organ
.	O

The	O
WNT	B-Gene_or_gene_product
/	O
TCF	B-Gene_or_gene_product
target	O
genes	O
HOXB9	B-Gene_or_gene_product
and	O
LEF1	B-Gene_or_gene_product
are	O
identified	O
as	O
mediators	O
of	O
chemotactic	O
invasion	O
and	O
colony	B-Cancer
outgrowth	O
.	O

Thus	O
,	O
a	O
distinct	O
WNT	B-Gene_or_gene_product
/	O
TCF	B-Gene_or_gene_product
signaling	O
program	O
through	O
LEF1	B-Gene_or_gene_product
and	O
HOXB9	B-Gene_or_gene_product
enhances	O
the	O
competence	O
of	O
lung	B-Cell
adenocarcinoma	I-Cell
cells	I-Cell
to	O
colonize	O
the	O
bones	B-Organ
and	O
the	O
brain	B-Organ
.	O

For	O
a	O
video	O
summary	O
of	O
this	O
article	O
,	O
see	O
the	O
PaperFlick	O
file	O
available	O
with	O
the	O
online	O
Supplemental	O
Data	O
.	O

Angiogenesis	O
-	O
a	O
novel	O
therapeutic	O
approach	O
for	O
ischemic	O
heart	B-Organ
disease	O
.	O

Angiogenesis	O
is	O
the	O
biologic	O
process	O
of	O
forming	O
new	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
.	O

Undoubtedly	O
,	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
growth	O
regulation	O
is	O
a	O
vital	O
aspect	O
in	O
health	O
and	O
disease	O
.	O

Under	O
physiological	O
conditions	O
,	O
angiogenesis	O
is	O
regulated	O
by	O
local	O
balance	O
between	O
endogenous	O
stimulators	O
and	O
inhibitors	O
of	O
this	O
process	O
.	O

In	O
many	O
diseases	O
state	O
body	B-Organism_subdivision
loses	O
control	O
over	O
angiogenesis	O
.	O

Angiogenesis	O
-	O
dependent	O
diseases	O
result	O
when	O
new	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
either	O
grow	O
excessively	O
or	O
insufficiently	O
.	O

Insufficient	O
angiogenesis	O
occurs	O
in	O
diseases	O
such	O
as	O
coronary	B-Multi-tissue_structure
artery	I-Multi-tissue_structure
disease	O
,	O
stroke	O
and	O
chronic	O
wounds	B-Pathological_formation
.	O

Myocardial	B-Multi-tissue_structure
ischemia	O
both	O
acute	O
and	O
chronic	O
has	O
been	O
clearly	O
shown	O
to	O
stimulate	O
angiogenesis	O
in	O
many	O
experimental	O
models	O
.	O

Therapeutic	O
angiogenesis	O
is	O
the	O
biological	O
agents	O
or	O
bioactive	O
material	O
to	O
stimulate	O
the	O
growth	O
of	O
new	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
.	O

Traditional	O
coronary	B-Multi-tissue_structure
revascularization	O
therapies	O
such	O
as	O
coronary	B-Multi-tissue_structure
angioplasty	O
or	O
bypass	B-Multi-tissue_structure
graft	I-Multi-tissue_structure
surgery	O
,	O
act	O
by	O
restoring	O
blood	B-Organism_substance
flow	O
through	O
the	O
preexisting	O
coronary	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
.	O

One	O
limitation	O
of	O
these	O
approaches	O
,	O
however	O
,	O
may	O
be	O
the	O
failure	O
to	O
normalize	O
myocardial	B-Multi-tissue_structure
perfusion	O
,	O
due	O
to	O
the	O
concomitant	O
presence	O
or	O
small	O
of	O
resistance	O
vessel	B-Multi-tissue_structure
disease	O
.	O

In	O
contrast	O
,	O
therapeutic	O
angiogenesis	O
is	O
based	O
on	O
the	O
concept	O
that	O
coronary	B-Multi-tissue_structure
collateral	I-Multi-tissue_structure
development	O
may	O
be	O
stimulated	O
by	O
pharmacological	O
or	O
molecular	O
means	O
and	O
can	O
limit	O
myocardial	B-Multi-tissue_structure
ischemia	O
.	O

Studies	O
,	O
both	O
in	O
human	B-Organism
and	O
animal	O
models	O
support	O
the	O
notion	O
that	O
,	O
various	O
angiogenic	O
growth	O
factors	O
and	O
progenitor	B-Cell
cells	I-Cell
can	O
enhance	O
new	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
.	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
fibroblast	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
FGF	B-Gene_or_gene_product
)	O
,	O
recombinant	O
proteins	O
and	O
bone	B-Cell
marrow	I-Cell
stem	I-Cell
cells	I-Cell
are	O
currently	O
used	O
therapeutic	O
stimulators	O
for	O
angiogenesis	O
.	O

As	O
coronary	B-Multi-tissue_structure
artery	I-Multi-tissue_structure
disease	O
is	O
the	O
major	O
cause	O
of	O
death	O
in	O
the	O
developed	O
societies	O
and	O
also	O
an	O
emerging	O
health	O
problem	O
in	O
developing	O
countries	O
like	O
Bangladesh	O
therapeutic	O
angiogenesis	O
may	O
provide	O
hope	O
as	O
a	O
new	O
treatment	O
modality	O
for	O
ischemic	O
heart	B-Organ
disease	O
with	O
or	O
in	O
place	O
of	O
current	O
therapies	O
.	O

Use	O
of	O
uteroglobin	B-Gene_or_gene_product
for	O
the	O
engineering	O
of	O
polyvalent	O
,	O
polyspecific	O
fusion	O
proteins	O
.	O

We	O
report	O
a	O
novel	O
strategy	O
to	O
engineer	O
and	O
express	O
stable	O
and	O
soluble	O
human	B-Organism
recombinant	O
polyvalent	O
/	O
polyspecific	O
fusion	O
proteins	O
.	O

The	O
procedure	O
is	O
based	O
on	O
the	O
use	O
of	O
a	O
central	O
skeleton	O
of	O
uteroglobin	B-Gene_or_gene_product
,	O
a	O
small	O
and	O
very	O
soluble	O
covalently	O
linked	O
homodimeric	O
protein	O
that	O
is	O
very	O
resistant	O
to	O
proteolytic	O
enzymes	O
and	O
to	O
pH	O
variations	O
.	O

Using	O
a	O
human	B-Organism
recombinant	O
antibody	O
(	O
scFv	O
)	O
specific	O
for	O
the	O
angiogenesis	O
marker	O
domain	O
B	O
of	O
fibronectin	B-Gene_or_gene_product
,	O
interleukin	B-Gene_or_gene_product
2	I-Gene_or_gene_product
,	O
and	O
an	O
scFv	O
able	O
to	O
neutralize	O
tumor	B-Gene_or_gene_product
necrosis	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
,	O
we	O
expressed	O
various	O
biologically	O
active	O
uteroglobin	B-Gene_or_gene_product
fusion	O
proteins	O
.	O

The	O
results	O
demonstrate	O
the	O
possibility	O
to	O
generate	O
monospecific	O
divalent	O
and	O
tetravalent	O
antibodies	O
,	O
immunocytokines	O
,	O
and	O
dual	O
specificity	O
tetravalent	O
antibodies	O
.	O

Furthermore	O
,	O
compared	O
with	O
similar	O
fusion	O
proteins	O
in	O
which	O
uteroglobin	B-Gene_or_gene_product
was	O
not	O
used	O
,	O
the	O
use	O
of	O
uteroglobin	B-Gene_or_gene_product
improved	O
properties	O
of	O
solubility	O
and	O
stability	O
.	O

Indeed	O
,	O
in	O
the	O
reported	O
cases	O
it	O
was	O
possible	O
to	O
vacuum	O
dry	O
and	O
reconstitute	O
the	O
proteins	O
without	O
any	O
aggregation	O
or	O
loss	O
in	O
protein	O
and	O
biological	O
activity	O
.	O

Netrin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
up	O
-	O
regulation	O
in	O
inflammatory	O
bowel	B-Organ
diseases	O
is	O
required	O
for	O
colorectal	B-Cancer
cancer	I-Cancer
progression	O
.	O

Chronic	O
inflammation	O
and	O
cancer	B-Cancer
are	O
intimately	O
associated	O
.	O

This	O
is	O
particularly	O
true	O
for	O
inflammatory	O
bowel	B-Organ
diseases	O
(	O
IBD	O
)	O
,	O
such	O
as	O
ulcerative	B-Pathological_formation
colitis	I-Pathological_formation
and	O
Crohn	O
'	O
s	O
disease	O
,	O
which	O
show	O
a	O
major	O
increased	O
risk	O
for	O
colorectal	B-Cancer
cancer	I-Cancer
.	O

While	O
the	O
understanding	O
of	O
the	O
molecular	O
pathogenesis	O
of	O
IBD	O
has	O
recently	O
improved	O
,	O
the	O
mechanisms	O
that	O
link	O
these	O
chronic	O
inflammatory	O
states	O
to	O
colorectal	B-Cancer
cancer	I-Cancer
development	O
are	O
in	O
large	O
part	O
unknown	O
.	O

One	O
of	O
these	O
mechanisms	O
is	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
pathway	O
activation	O
which	O
in	O
turn	O
may	O
contribute	O
to	O
tumor	B-Cancer
formation	O
by	O
providing	O
anti	O
-	O
apoptotic	O
survival	O
signals	O
to	O
the	O
epithelial	B-Cell
cells	I-Cell
.	O

Based	O
on	O
the	O
observation	O
that	O
netrin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
the	O
anti	O
-	O
apoptotic	O
ligand	O
for	O
the	O
dependence	O
receptors	O
DCC	B-Gene_or_gene_product
and	O
UNC5H	B-Gene_or_gene_product
is	O
up	O
-	O
regulated	O
in	O
colonic	B-Organ
crypts	I-Organ
in	O
response	O
to	O
NF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kappaB	I-Gene_or_gene_product
,	O
we	O
show	O
here	O
that	O
colorectal	B-Cancer
cancers	I-Cancer
from	O
inflammatory	O
bowel	B-Organ
diseases	O
patients	B-Organism
have	O
selected	O
up	O
-	O
regulation	O
of	O
netrin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

Moreover	O
,	O
we	O
demonstrate	O
that	O
this	O
inflammation	O
-	O
driven	O
netrin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
up	O
-	O
regulation	O
is	O
causal	O
for	O
colorectal	B-Cancer
cancer	I-Cancer
development	O
as	O
interference	O
with	O
netrin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
autocrine	O
loop	O
in	O
a	O
mouse	B-Organism
model	O
for	O
ulcerative	B-Pathological_formation
colitis	O
-	O
associated	O
colorectal	B-Cancer
cancer	I-Cancer
,	O
while	O
showing	O
no	O
effect	O
on	O
inflammation	O
,	O
inhibits	O
colorectal	B-Cancer
cancer	I-Cancer
progression	O
.	O

[	O
Prognostic	O
significance	O
of	O
cellular	B-Cell
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
in	O
the	O
course	O
of	O
chronic	B-Cancer
myeloid	I-Cancer
leukaemia	I-Cancer
]	O

INTRODUCTION	O
:	O
Increased	O
angiogenesis	O
in	O
bone	B-Multi-tissue_structure
marrow	I-Multi-tissue_structure
is	O
one	O
of	O
the	O
characteristics	O
of	O
chronic	B-Cancer
myeloid	I-Cancer
leukaemia	I-Cancer
(	O
CML	B-Cancer
)	O
,	O
a	O
clonal	O
myeloproliferative	O
disorder	O
that	O
expresses	O
a	O
chimeric	O
bcr	B-Gene_or_gene_product
/	I-Gene_or_gene_product
abl	I-Gene_or_gene_product
protein	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
potent	O
and	O
a	O
specific	O
regulator	O
of	O
angiogenesis	O
which	O
principally	O
targets	O
endothelial	B-Cell
cells	I-Cell
and	O
regulates	O
several	O
of	O
their	O
functions	O
,	O
including	O
mitogenesis	O
,	O
permeability	O
and	O
migration	O
.	O

The	O
impact	O
of	O
elevated	O
VEGF	B-Gene_or_gene_product
expression	O
on	O
the	O
course	O
of	O
chronic	B-Cancer
myeloid	I-Cancer
leukaemia	I-Cancer
is	O
unknown	O
.	O

OBJECTIVE	O
:	O
The	O
aim	O
of	O
this	O
study	O
was	O
the	O
follow	O
-	O
up	O
of	O
VEGF	B-Gene_or_gene_product
expression	O
during	O
the	O
course	O
of	O
CML	B-Cancer
.	O

METHODS	O
:	O
We	O
studied	O
VEGF	B-Gene_or_gene_product
expression	O
of	O
85	O
CML	B-Cancer
patients	B-Organism
(	O
median	O
age	O
50	O
years	O
,	O
range	O
16	O
-	O
75	O
years	O
)	O
.	O

At	O
the	O
commencement	O
of	O
the	O
study	O
,	O
29	O
patients	B-Organism
were	O
in	O
chronic	O
phase	O
(	O
CP	O
)	O
,	O
25	O
in	O
an	O
accelerated	O
phase	O
(	O
AP	O
)	O
,	O
and	O
31	O
in	O
the	O
blast	O
crisis	O
(	O
BC	O
)	O
.	O

The	O
temporal	O
expression	O
(	O
percentage	O
positivity	O
per	O
1000	O
analysed	O
cells	B-Cell
)	O
VEGF	B-Gene_or_gene_product
proteins	O
over	O
the	O
course	O
of	O
CML	B-Cancer
were	O
studied	O
using	O
the	O
immunohistochemical	O
technique	O
utilizing	O
relevant	O
monoclonal	O
antibodies	O
.	O

It	O
was	O
correlated	O
with	O
the	O
laboratory	O
(	O
Hb	B-Gene_or_gene_product
,	O
WBC	B-Cell
and	O
platelet	B-Cell
counts	O
,	O
and	O
the	O
percentage	O
of	O
blasts	O
)	O
and	O
clinical	O
parameters	O
(	O
organomegaly	O
,	O
duration	O
of	O
CP	O
,	O
AP	O
,	O
and	O
BC	O
)	O
of	O
disease	O
progression	O
.	O

RESULTS	O
:	O
The	O
expression	O
ofVEGF	B-Gene_or_gene_product
protein	O
was	O
most	O
pronounced	O
in	O
AP	O
(	O
ANOVA	O
,	O
p	O
=	O
0	O
.	O
033	O
)	O
.	O

The	O
level	O
of	O
VEGF	B-Gene_or_gene_product
expression	O
correlated	O
inversely	O
with	O
the	O
degree	O
of	O
splenomegaly	B-Pathological_formation
(	O
Pearson	O
,	O
r	O
=	O
-	O
0	O
.	O
400	O
,	O
p	O
=	O
0	O
.	O
011	O
)	O
.	O

High	O
expression	O
of	O
VEGF	B-Gene_or_gene_product
correlated	O
with	O
a	O
shorter	O
overall	O
survival	O
(	O
log	O
rank	O
,	O
p	O
=	O
0	O
.	O
042	O
)	O
.	O

CONCLUSION	O
:	O
Immunohistochemically	O
confirmed	O
significance	O
of	O
the	O
expression	O
of	O
VEGF	B-Gene_or_gene_product
in	O
dependence	O
of	O
the	O
CML	B-Cancer
stage	O
could	O
be	O
of	O
clinical	O
importance	O
in	O
deciding	O
on	O
the	O
timing	O
therapy	O
.	O

These	O
data	O
suggest	O
that	O
VEGF	B-Gene_or_gene_product
plays	O
a	O
role	O
in	O
the	O
biology	O
of	O
CML	B-Cancer
and	O
that	O
VEGF	B-Gene_or_gene_product
inhibitors	O
should	O
be	O
investigated	O
in	O
CML	B-Cancer
.	O

Pbx1	B-Gene_or_gene_product
is	O
a	O
downstream	O
target	O
of	O
Evi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
in	O
hematopoietic	B-Cell
stem	I-Cell
/	O
progenitors	B-Cell
and	O
leukemic	B-Cell
cells	I-Cell
.	O

Ecotropic	B-Gene_or_gene_product
viral	I-Gene_or_gene_product
integration	I-Gene_or_gene_product
site	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
Evi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
is	O
a	O
nuclear	B-Cellular_component
transcription	O
factor	O
,	O
which	O
is	O
essential	O
for	O
the	O
proliferation	O
/	O
maintenance	O
of	O
hematopoietic	B-Cell
stem	I-Cell
cells	I-Cell
(	O
HSCs	B-Cell
)	O
.	O

Aberrant	O
expression	O
of	O
Evi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
has	O
been	O
frequently	O
found	O
in	O
myeloid	B-Cancer
leukemia	I-Cancer
,	O
and	O
is	O
associated	O
with	O
a	O
poor	O
patient	B-Organism
survival	O
.	O

Recently	O
,	O
we	O
reported	O
candidate	O
target	O
genes	O
of	O
Evi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
shared	O
in	O
HSCs	B-Cell
and	O
leukemic	B-Cell
cells	I-Cell
using	O
gene	O
expression	O
profiling	O
analysis	O
.	O

In	O
this	O
study	O
,	O
we	O
identified	O
Pbx1	B-Gene_or_gene_product
,	O
a	O
proto	O
-	O
oncogene	O
in	O
hematopoietic	B-Pathological_formation
malignancy	I-Pathological_formation
,	O
as	O
a	O
target	O
gene	O
of	O
Evi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

Overexpression	O
of	O
Evi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
increased	O
Pbx1	B-Gene_or_gene_product
expression	O
in	O
hematopoietic	B-Cell
stem	I-Cell
/	O
progenitor	B-Cell
cells	I-Cell
.	O

An	O
analysis	O
of	O
the	O
Pbx1	B-Gene_or_gene_product
promoter	O
region	O
revealed	O
that	O
Evi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
upregulates	O
Pbx1	B-Gene_or_gene_product
transcription	O
.	O

Furthermore	O
,	O
reduction	O
of	O
Pbx1	B-Gene_or_gene_product
levels	O
through	O
RNAi	O
-	O
mediated	O
knockdown	O
significantly	O
inhibited	O
Evi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
induced	O
transformation	O
.	O

In	O
contrast	O
,	O
knockdown	O
of	O
Pbx1	B-Gene_or_gene_product
did	O
not	O
impair	O
bone	B-Multi-tissue_structure
marrow	I-Multi-tissue_structure
transformation	O
by	O
E2A	B-Gene_or_gene_product
/	O
HLF	B-Gene_or_gene_product
or	O
AML1	B-Gene_or_gene_product
/	O
ETO	B-Gene_or_gene_product
,	O
suggesting	O
that	O
Pbx1	B-Gene_or_gene_product
is	O
specifically	O
required	O
for	O
the	O
maintenance	O
of	O
bone	B-Multi-tissue_structure
marrow	I-Multi-tissue_structure
transformation	O
mediated	O
by	O
Evi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
.	O

These	O
results	O
indicate	O
that	O
Pbx1	B-Gene_or_gene_product
is	O
a	O
target	O
gene	O
of	O
Evi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
involved	O
in	O
Evi	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
mediated	O
leukemogenesis	O
.	O

Tumor	B-Cancer
Angiogenesis	O
:	O
Initiation	O
and	O
Targeting	O
-	O
Therapeutic	O
Targeting	O
of	O
an	O
FGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Binding	I-Gene_or_gene_product
Protein	I-Gene_or_gene_product
,	O
an	O
Angiogenic	O
Switch	O
Molecule	O
,	O
and	O
Indicator	O
of	O
Early	O
Stages	O
of	O
Gastrointestinal	B-Cancer
Adenocarcinomas	I-Cancer
-	O
.	O

Tumor	B-Cancer
angiogenesis	O
has	O
been	O
related	O
to	O
the	O
initiation	O
as	O
well	O
as	O
progression	O
toward	O
more	O
aggressive	O
behavior	O
of	O
human	B-Organism
tumors	B-Cancer
.	O

In	O
particular	O
,	O
the	O
activity	O
of	O
angiogenic	O
factors	O
is	O
crucial	O
for	O
tumor	B-Cancer
progression	O
.	O

We	O
previously	O
characterized	O
a	O
secreted	O
fibroblast	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
binding	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
(	O
FGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
BP	I-Gene_or_gene_product
)	O
as	O
a	O
chaperone	O
molecule	O
,	O
which	O
binds	O
to	O
various	O
FGFs	B-Gene_or_gene_product
,	O
enhances	O
FGF	B-Gene_or_gene_product
-	O
mediated	O
biochemical	O
and	O
biologic	O
events	O
and	O
importantly	O
is	O
a	O
crucial	O
rate	O
-	O
limiting	O
factor	O
for	O
tumor	B-Cancer
-	O
dependent	O
angiogenesis	O
.	O

We	O
generated	O
monoclonal	O
antibodies	O
that	O
target	O
FGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
BP	I-Gene_or_gene_product
protein	O
and	O
used	O
them	O
as	O
a	O
tool	O
to	O
evaluate	O
frequency	O
and	O
pattern	O
of	O
FGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
BP	I-Gene_or_gene_product
expression	O
during	O
the	O
malignant	O
progression	O
of	O
pancreas	B-Cancer
and	O
colorectal	B-Cancer
carcinoma	I-Cancer
in	O
archival	O
tissue	B-Tissue
samples	I-Tissue
.	O

We	O
found	O
that	O
FGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
BP	I-Gene_or_gene_product
is	O
dramatically	O
upregulated	O
during	O
the	O
initiation	O
of	O
colorectal	B-Cancer
and	O
pancreatic	B-Cancer
adenocarcinoma	I-Cancer
.	O

Crucial	O
genetic	O
events	O
underlying	O
the	O
initiation	O
and	O
progression	O
of	O
colorectal	B-Cancer
and	O
pancreatic	B-Cancer
adenocarcinoma	I-Cancer
with	O
a	O
particular	O
focus	O
on	O
the	O
modulation	O
of	O
angiogenesis	O
and	O
antiangiogenic	O
therapies	O
are	O
discussed	O
.	O

We	O
propose	O
that	O
the	O
upregulation	O
of	O
the	O
secreted	O
FGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
BP	I-Gene_or_gene_product
protein	O
during	O
early	O
phases	O
of	O
pancreas	B-Cancer
and	O
colon	B-Cancer
cancer	I-Cancer
could	O
make	O
this	O
protein	O
a	O
possible	O
serum	B-Organism_substance
marker	O
indicating	O
the	O
presence	O
of	O
high	O
-	O
risk	O
premalignant	B-Pathological_formation
lesions	I-Pathological_formation
.	O

Furthermore	O
,	O
the	O
biological	O
activity	O
of	O
FGF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
BP	I-Gene_or_gene_product
is	O
neutralized	O
by	O
monoclonal	O
antibodies	O
suggesting	O
the	O
potential	O
for	O
antibody	O
-	O
based	O
therapeutic	O
targeting	O
.	O

The	O
role	O
of	O
hypoxia	O
in	O
2	B-Simple_chemical
-	I-Simple_chemical
butoxyethanol	I-Simple_chemical
-	O
induced	O
hemangiosarcoma	B-Cancer
.	O

To	O
understand	O
the	O
molecular	O
mechanisms	O
underlying	O
compound	O
-	O
induced	O
hemangiosarcomas	B-Cancer
in	O
mice	B-Organism
,	O
and	O
therefore	O
,	O
their	O
human	B-Organism
relevance	O
,	O
a	O
systems	O
biology	O
approach	O
was	O
undertaken	O
using	O
transcriptomics	O
and	O
Causal	O
Network	O
Modeling	O
from	O
mice	B-Organism
treated	O
with	O
2	B-Simple_chemical
-	I-Simple_chemical
butoxyethanol	I-Simple_chemical
(	O
2	B-Simple_chemical
-	I-Simple_chemical
BE	I-Simple_chemical
)	O
.	O

2	B-Simple_chemical
-	I-Simple_chemical
BE	I-Simple_chemical
is	O
a	O
hemolytic	O
agent	O
that	O
induces	O
hemangiosarcomas	B-Cancer
in	O
mice	B-Organism
.	O

We	O
hypothesized	O
that	O
the	O
hemolysis	O
induced	O
by	O
2	B-Simple_chemical
-	I-Simple_chemical
BE	I-Simple_chemical
would	O
result	O
in	O
local	O
tissue	B-Tissue
hypoxia	O
,	O
a	O
well	O
-	O
documented	O
trigger	O
for	O
endothelial	B-Cell
cell	I-Cell
proliferation	O
leading	O
to	O
hemangiosarcoma	B-Cancer
.	O

Gene	O
expression	O
data	O
from	O
bone	B-Multi-tissue_structure
marrow	I-Multi-tissue_structure
(	O
BM	B-Multi-tissue_structure
)	O
,	O
liver	B-Organ
,	O
and	O
spleen	B-Organ
of	O
mice	B-Organism
exposed	O
to	O
a	O
single	O
dose	O
(	O
4	O
h	O
)	O
or	O
seven	O
daily	O
doses	O
of	O
2	B-Simple_chemical
-	I-Simple_chemical
BE	I-Simple_chemical
were	O
used	O
to	O
develop	O
a	O
mechanistic	O
model	O
of	O
hemangiosarcoma	B-Cancer
.	O

The	O
resulting	O
mechanistic	O
model	O
confirms	O
previous	O
work	O
proposing	O
that	O
2	B-Simple_chemical
-	I-Simple_chemical
BE	I-Simple_chemical
induces	O
macrophage	B-Cell
activation	O
and	O
inflammation	O
in	O
the	O
liver	B-Organ
.	O

In	O
addition	O
,	O
the	O
model	O
supports	O
local	O
tissue	B-Tissue
hypoxia	O
in	O
the	O
liver	B-Organ
and	O
spleen	B-Organ
,	O
coupled	O
with	O
increased	O
erythropoeitin	B-Gene_or_gene_product
signaling	O
and	O
erythropoiesis	O
in	O
the	O
spleen	B-Organ
and	O
BM	B-Multi-tissue_structure
,	O
and	O
suppression	O
of	O
mechanisms	O
that	O
contribute	O
to	O
genomic	O
stability	O
,	O
events	O
that	O
could	O
be	O
contributing	O
factors	O
to	O
hemangiosarcoma	B-Cancer
formation	O
.	O

Finally	O
,	O
an	O
immunohistochemistry	O
method	O
(	O
Hypoxyprobe	O
)	O
demonstrated	O
that	O
tissue	B-Tissue
hypoxia	O
was	O
present	O
in	O
the	O
spleen	B-Organ
and	O
BM	B-Multi-tissue_structure
.	O

Together	O
,	O
the	O
results	O
of	O
this	O
study	O
identify	O
molecular	O
mechanisms	O
that	O
initiate	O
hemangiosarcoma	B-Cancer
,	O
a	O
key	O
step	O
in	O
understanding	O
safety	O
concerns	O
that	O
can	O
impact	O
drug	O
decision	O
processes	O
,	O
and	O
identified	O
hypoxia	O
as	O
a	O
possible	O
contributing	O
factor	O
for	O
2	B-Simple_chemical
-	I-Simple_chemical
BE	I-Simple_chemical
-	O
induced	O
hemangiosarcoma	B-Cancer
in	O
mice	B-Organism
.	O

Pancreatic	B-Cancer
endocrine	I-Cancer
tumors	I-Cancer
:	O
expression	O
profiling	O
evidences	O
a	O
role	O
for	O
AKT	B-Gene_or_gene_product
-	O
mTOR	B-Gene_or_gene_product
pathway	O
.	O

PURPOSE	O
:	O
We	O
investigated	O
the	O
global	O
gene	O
expression	O
in	O
a	O
large	O
panel	O
of	O
pancreatic	B-Cancer
endocrine	I-Cancer
tumors	I-Cancer
(	O
PETs	B-Cancer
)	O
aimed	O
at	O
identifying	O
new	O
potential	O
targets	O
for	O
therapy	O
and	O
biomarkers	O
to	O
predict	O
patient	B-Organism
outcome	O
.	O

PATIENTS	O
AND	O
METHODS	O
:	O
Using	O
a	O
custom	O
microarray	O
,	O
we	O
analyzed	O
72	O
primary	O
PETs	B-Cancer
,	O
seven	O
matched	O
metastases	O
,	O
and	O
10	O
normal	O
pancreatic	B-Multi-tissue_structure
samples	I-Multi-tissue_structure
.	O

Relevant	O
differentially	O
expressed	O
genes	O
were	O
validated	O
by	O
either	O
quantitative	O
real	O
-	O
time	O
polymerase	O
chain	O
reaction	O
or	O
immunohistochemistry	O
on	O
tissue	B-Tissue
microarrays	O
.	O

RESULTS	O
:	O
Our	O
data	O
showed	O
that	O
:	O
tuberous	B-Gene_or_gene_product
sclerosis	I-Gene_or_gene_product
2	I-Gene_or_gene_product
(	O
TSC2	B-Gene_or_gene_product
)	O
and	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
(	O
PTEN	B-Gene_or_gene_product
)	O
were	O
downregulated	O
in	O
most	O
of	O
the	O
primary	O
tumors	B-Cancer
,	O
and	O
their	O
low	O
expression	O
was	O
significantly	O
associated	O
with	O
shorter	O
disease	O
-	O
free	O
and	O
overall	O
survival	O
;	O
somatostatin	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
2	I-Gene_or_gene_product
(	O
SSTR2	B-Gene_or_gene_product
)	O
was	O
absent	O
or	O
very	O
low	O
in	O
insulinomas	B-Cancer
compared	O
with	O
nonfunctioning	O
tumors	B-Cancer
;	O
and	O
expression	O
of	O
fibroblast	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
13	I-Gene_or_gene_product
(	O
FGF13	B-Gene_or_gene_product
)	O
gene	O
was	O
significantly	O
associated	O
with	O
the	O
occurrence	O
of	O
liver	B-Organ
metastasis	O
and	O
shorter	O
disease	O
-	O
free	O
survival	O
.	O

TSC2	B-Gene_or_gene_product
and	O
PTEN	B-Gene_or_gene_product
are	O
two	O
key	O
inhibitors	O
of	O
the	O
Akt	B-Gene_or_gene_product
/	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
pathway	O
and	O
the	O
specific	O
inhibition	O
of	O
mTOR	B-Gene_or_gene_product
with	O
rapamycin	B-Simple_chemical
or	O
RAD001	B-Simple_chemical
inhibited	O
cell	B-Cell
proliferation	O
of	O
PET	B-Cell
cell	I-Cell
lines	I-Cell
.	O

CONCLUSION	O
:	O
Our	O
results	O
strongly	O
support	O
a	O
role	O
for	O
PI3K	B-Gene_or_gene_product
/	O
Akt	B-Gene_or_gene_product
/	O
mTOR	B-Gene_or_gene_product
pathway	O
in	O
PET	B-Cancer
,	O
which	O
ties	O
in	O
with	O
the	O
fact	O
that	O
mTOR	B-Gene_or_gene_product
inhibitors	O
have	O
reached	O
phase	O
III	O
trials	O
in	O
neuroendocrine	B-Cancer
tumors	I-Cancer
.	O

The	O
finding	O
of	O
differential	O
SSTR	B-Gene_or_gene_product
expression	O
raises	O
the	O
potential	O
for	O
SSTR	B-Gene_or_gene_product
expression	O
to	O
be	O
evaluated	O
as	O
a	O
marker	O
of	O
response	O
to	O
somatostatin	B-Gene_or_gene_product
analogs	O
.	O

Finally	O
,	O
we	O
identified	O
FGF13	B-Gene_or_gene_product
as	O
a	O
new	O
prognostic	O
marker	O
that	O
predicted	O
poorer	O
outcome	O
in	O
patients	B-Organism
who	O
were	O
clinically	O
considered	O
free	O
from	O
disease	O
.	O

ICAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
enhances	O
the	O
migratory	O
and	O
invasive	O
potential	O
of	O
human	B-Organism
non	B-Cell
-	I-Cell
small	I-Cell
cell	I-Cell
lung	I-Cell
cancer	I-Cell
cells	I-Cell
by	O
inducing	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
via	O
Akt	B-Gene_or_gene_product
and	O
CREB	B-Gene_or_gene_product
.	O

We	O
have	O
previously	O
reported	O
that	O
intercellular	B-Gene_or_gene_product
adhesion	I-Gene_or_gene_product
molecule	I-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
(	O
ICAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
)	O
is	O
associated	O
with	O
an	O
increase	O
of	O
cellular	B-Cell
radio	O
-	O
resistance	O
and	O
cancer	B-Cell
cell	I-Cell
proliferation	O
.	O

In	O
this	O
study	O
,	O
we	O
hypothesized	O
that	O
ICAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
has	O
an	O
additional	O
effect	O
on	O
cancer	B-Cell
cell	I-Cell
migration	O
and	O
invasion	O
because	O
molecules	O
induced	O
by	O
ICAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
are	O
known	O
as	O
regulators	O
of	O
cell	B-Cell
migration	O
and	O
invasion	O
.	O

To	O
examine	O
this	O
hypothesis	O
,	O
we	O
used	O
NCI	B-Cell
-	I-Cell
H1299	I-Cell
non	I-Cell
-	I-Cell
small	I-Cell
cell	I-Cell
lung	I-Cell
cancer	I-Cell
(	I-Cell
NSCLC	I-Cell
)	I-Cell
cell	I-Cell
line	I-Cell
(	O
p53	B-Gene_or_gene_product
and	O
PTEN	B-Gene_or_gene_product
null	O
cell	O
)	O
and	O
constructed	O
an	O
ICAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
-	O
over	O
-	O
expressing	O
stable	O
transfectant	B-Cell
,	O
which	O
exhibited	O
increased	O
cell	B-Cell
migration	O
and	O
invasion	O
.	O

The	O
increased	O
migration	O
and	O
invasion	O
resulted	O
from	O
up	O
-	O
regulation	O
of	O
expression	O
and	O
activities	O
of	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
.	O

ICAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
also	O
increased	O
Akt	B-Gene_or_gene_product
phosphorylation	O
,	O
which	O
caused	O
an	O
increase	O
in	O
cellular	B-Cell
migration	O
/	O
invasion	O
and	O
MMP	B-Gene_or_gene_product
activities	O
.	O

Activity	O
of	O
several	O
transcriptional	O
factors	O
located	O
downstream	O
in	O
the	O
Akt	B-Gene_or_gene_product
pathway	O
was	O
also	O
tested	O
,	O
and	O
constitutive	O
activation	O
of	O
adenosine	B-Gene_or_gene_product
3	I-Gene_or_gene_product
'	I-Gene_or_gene_product
,	I-Gene_or_gene_product
5	I-Gene_or_gene_product
'	I-Gene_or_gene_product
-	I-Gene_or_gene_product
monophosphate	I-Gene_or_gene_product
response	I-Gene_or_gene_product
element	I-Gene_or_gene_product
-	I-Gene_or_gene_product
binding	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
(	O
CREB	B-Gene_or_gene_product
)	O
by	O
ICAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
was	O
detected	O
.	O

Blockage	O
of	O
the	O
Akt	B-Gene_or_gene_product
pathway	O
attenuated	O
CREB	B-Gene_or_gene_product
activation	O
,	O
and	O
a	O
decrease	O
in	O
CREB	B-Gene_or_gene_product
expression	O
reduced	O
cellular	B-Cell
migration	O
/	O
invasion	O
and	O
activity	O
of	O
MMPs	B-Gene_or_gene_product
.	O

This	O
result	O
indicates	O
that	O
CREB	B-Gene_or_gene_product
functions	O
in	O
the	O
signaling	O
pathway	O
between	O
Akt	B-Gene_or_gene_product
and	O
MMP	B-Gene_or_gene_product
.	O

We	O
also	O
showed	O
ICAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
-	O
induced	O
cell	B-Cell
migration	O
and	O
invasion	O
in	O
NCI	B-Cell
-	I-Cell
H460	I-Cell
NSCLC	I-Cell
cells	I-Cell
(	O
wild	O
-	O
type	O
p53	B-Gene_or_gene_product
and	O
PTEN	B-Gene_or_gene_product
cell	O
)	O
through	O
the	O
same	O
signaling	O
pathway	O
.	O

Taken	O
together	O
,	O
our	O
findings	O
suggest	O
that	O
ICAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
stimulates	O
cancer	B-Cell
cell	I-Cell
migration	O
/	O
invasion	O
via	O
ICAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
/	O
Akt	B-Gene_or_gene_product
/	O
CREB	B-Gene_or_gene_product
/	O
MMP	B-Gene_or_gene_product
pathway	O
regardless	O
of	O
p53	B-Gene_or_gene_product
and	O
PTEN	B-Gene_or_gene_product
status	O
,	O
and	O
this	O
reflects	O
the	O
possibility	O
that	O
ICAM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
could	O
be	O
considered	O
as	O
a	O
candidate	O
for	O
anti	O
-	O
cancer	B-Cancer
drug	O
development	O
and	O
as	O
a	O
cancer	B-Cancer
diagnostic	O
marker	O
.	O

The	O
role	O
of	O
p53	B-Gene_or_gene_product
in	O
glucose	B-Simple_chemical
metabolism	O
.	O

The	O
p53	B-Gene_or_gene_product
protein	O
functions	O
to	O
prevent	O
tumour	B-Cancer
development	O
by	O
inhibiting	O
the	O
outgrowth	O
of	O
stressed	O
or	O
damaged	O
cells	B-Cell
.	O

In	O
addition	O
to	O
well	O
established	O
functions	O
to	O
block	O
cell	B-Cell
proliferation	O
,	O
recent	O
studies	O
have	O
revealed	O
a	O
role	O
for	O
p53	B-Gene_or_gene_product
in	O
the	O
regulation	O
of	O
pathways	O
involved	O
in	O
glucose	B-Simple_chemical
metabolism	O
.	O

The	O
metabolic	O
functions	O
of	O
p53	B-Gene_or_gene_product
resist	O
the	O
shift	O
to	O
glycolysis	O
that	O
is	O
characteristically	O
seen	O
in	O
cancers	B-Cancer
,	O
and	O
also	O
help	O
cells	B-Cell
adapt	O
to	O
and	O
survive	O
limited	O
periods	O
of	O
metabolic	O
stress	O
.	O

Such	O
activities	O
of	O
p53	B-Gene_or_gene_product
would	O
not	O
only	O
help	O
to	O
prevent	O
cancer	B-Cancer
development	O
,	O
but	O
might	O
also	O
contribute	O
to	O
non	O
-	O
tumour	B-Cancer
related	O
roles	O
for	O
p53	B-Gene_or_gene_product
,	O
such	O
as	O
in	O
the	O
regulation	O
of	O
longevity	O
.	O

These	O
new	O
functions	O
of	O
p53	B-Gene_or_gene_product
are	O
providing	O
interesting	O
possibilities	O
for	O
the	O
development	O
of	O
novel	O
therapies	O
.	O

14	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
-	I-Gene_or_gene_product
3zeta	I-Gene_or_gene_product
Overexpression	O
and	O
abnormal	O
beta	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
expression	O
are	O
associated	O
with	O
poor	O
differentiation	O
and	O
progression	O
in	O
stage	B-Cancer
I	I-Cancer
non	I-Cancer
-	I-Cancer
small	I-Cancer
cell	I-Cancer
lung	I-Cancer
cancer	I-Cancer
.	O

Recent	O
studies	O
have	O
shown	O
that	O
14	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
-	I-Gene_or_gene_product
3zeta	I-Gene_or_gene_product
interacted	O
with	O
other	O
key	O
cellular	B-Cell
proteins	O
involved	O
in	O
the	O
tumor	B-Cancer
development	O
and	O
progression	O
.	O

Knowledge	O
of	O
14	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
-	I-Gene_or_gene_product
3zeta	I-Gene_or_gene_product
and	O
beta	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
expression	O
and	O
clinical	O
significance	O
in	O
the	O
same	B-Tissue
tumor	I-Tissue
tissues	I-Tissue
is	O
limited	O
.	O

The	O
purpose	O
of	O
this	O
study	O
was	O
to	O
investigate	O
the	O
expression	O
and	O
significance	O
of	O
14	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
-	I-Gene_or_gene_product
3zeta	I-Gene_or_gene_product
and	O
beta	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
in	O
stage	O
I	O
non	B-Cancer
-	I-Cancer
small	I-Cancer
-	I-Cancer
cell	I-Cancer
lung	I-Cancer
cancer	I-Cancer
(	O
NSCLC	B-Cancer
)	O
.	O

Specimens	O
of	O
NSCLC	B-Cancer
and	O
adjacent	O
normal	B-Tissue
lung	I-Tissue
tissues	I-Tissue
were	O
collected	O
from	O
110	O
patients	B-Organism
.	O

The	O
expressions	O
of	O
14	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
-	I-Gene_or_gene_product
3zeta	I-Gene_or_gene_product
and	O
beta	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
were	O
detected	O
by	O
western	O
blotting	O
,	O
double	O
labeling	O
immunofluorescence	O
,	O
confocal	O
laser	O
scanning	O
microscopy	O
and	O
immunohistochemistry	O
.	O

The	O
expression	O
of	O
14	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
-	I-Gene_or_gene_product
3zeta	I-Gene_or_gene_product
was	O
upregulated	O
in	O
stage	B-Cancer
I	I-Cancer
NSCLC	I-Cancer
.	O

Further	O
,	O
the	O
overexpression	O
of	O
14	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
-	I-Gene_or_gene_product
3zeta	I-Gene_or_gene_product
correlated	O
with	O
histological	O
grades	O
,	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
and	O
poor	O
clinical	O
outcome	O
.	O

Abnormal	O
expression	O
of	O
beta	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
was	O
significantly	O
correlated	O
with	O
poor	O
differentiation	O
and	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
.	O

Abnormal	O
beta	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
expression	O
was	O
associated	O
significantly	O
with	O
positive	O
14	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
-	I-Gene_or_gene_product
3zeta	I-Gene_or_gene_product
expression	O
.	O

In	O
conclusion	O
,	O
14	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
-	I-Gene_or_gene_product
3zeta	I-Gene_or_gene_product
and	O
beta	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
might	O
have	O
an	O
important	O
role	O
in	O
development	O
,	O
progression	O
and	O
metastatic	O
process	O
of	O
NSCLC	B-Cancer
.	O

14	B-Gene_or_gene_product
-	I-Gene_or_gene_product
3	I-Gene_or_gene_product
-	I-Gene_or_gene_product
3zeta	I-Gene_or_gene_product
might	O
be	O
used	O
as	O
prognostic	O
biomarkers	O
for	O
NSCLC	B-Cancer
.	O

p24	B-Gene_or_gene_product
family	I-Gene_or_gene_product
type	I-Gene_or_gene_product
1	I-Gene_or_gene_product
transmembrane	I-Gene_or_gene_product
proteins	I-Gene_or_gene_product
are	O
required	O
for	O
insulin	B-Gene_or_gene_product
biosynthesis	O
and	O
secretion	O
in	O
pancreatic	B-Cell
beta	I-Cell
-	I-Cell
cells	I-Cell
.	O

The	O
p24	B-Gene_or_gene_product
protein	I-Gene_or_gene_product
family	I-Gene_or_gene_product
have	O
multiple	O
functions	O
in	O
protein	O
transport	O
in	O
the	O
early	O
secretory	O
pathway	O
.	O

In	O
this	O
study	O
we	O
examined	O
the	O
role	O
of	O
p24	B-Gene_or_gene_product
proteins	I-Gene_or_gene_product
in	O
insulin	B-Gene_or_gene_product
transport	O
.	O

Several	O
members	O
were	O
detected	O
in	O
insulinoma	B-Cell
cell	I-Cell
lines	I-Cell
and	O
rat	B-Organism
islets	B-Multi-tissue_structure
and	O
expression	O
levels	O
positively	O
correlated	O
with	O
insulin	B-Gene_or_gene_product
abundance	O
,	O
particularly	O
for	O
p24delta1	B-Gene_or_gene_product
and	O
p24beta1	B-Gene_or_gene_product
.	O

Knocking	O
down	O
p24delta1	B-Gene_or_gene_product
in	O
insulinoma	B-Cell
cell	I-Cell
lines	I-Cell
,	O
which	O
also	O
resulted	O
in	O
the	O
concomitant	O
knock	O
-	O
down	O
of	O
other	O
family	O
members	O
,	O
impaired	O
glucose	B-Simple_chemical
-	O
stimulated	O
insulin	B-Gene_or_gene_product
secretion	O
,	O
decreased	O
total	O
cellular	B-Cell
insulin	B-Gene_or_gene_product
content	O
and	O
reduced	O
proinsulin	B-Gene_or_gene_product
biosynthesis	O
.	O

There	O
was	O
no	O
effect	O
on	O
overall	O
protein	O
biosynthesis	O
or	O
ER	B-Gene_or_gene_product
stress	O
.	O

These	O
results	O
suggest	O
that	O
p24delta1	B-Gene_or_gene_product
and	O
possibly	O
other	O
p24	B-Gene_or_gene_product
family	I-Gene_or_gene_product
proteins	I-Gene_or_gene_product
are	O
required	O
for	O
normal	O
insulin	B-Gene_or_gene_product
biosynthesis	O
and	O
subsequent	O
secretion	O
in	O
pancreatic	B-Cell
beta	I-Cell
-	I-Cell
cells	I-Cell
.	O

Expression	O
of	O
KISS1	B-Gene_or_gene_product
and	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
in	O
non	B-Cancer
-	I-Cancer
small	I-Cancer
cell	I-Cancer
lung	I-Cancer
cancer	I-Cancer
and	O
their	O
relations	O
to	O
metastasis	O
and	O
survival	O
.	O

KISS1	B-Gene_or_gene_product
and	O
matrix	B-Gene_or_gene_product
metalloproteinase	I-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
(	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
)	O
may	O
play	O
important	O
roles	O
as	O
metastasis	O
suppressor	O
and	O
metastasis	O
promoter	O
genes	O
,	O
respectively	O
,	O
in	O
a	O
variety	O
of	O
malignancies	B-Cancer
.	O

However	O
,	O
there	O
is	O
little	O
information	O
about	O
their	O
possible	O
roles	O
in	O
non	B-Cancer
-	I-Cancer
small	I-Cancer
cell	I-Cancer
lung	I-Cancer
cancer	I-Cancer
(	O
NSCLC	B-Cancer
)	O
.	O

The	O
goals	O
of	O
this	O
study	O
were	O
to	O
determine	O
the	O
mRNA	O
and	O
protein	O
expressions	O
of	O
KISS1	B-Gene_or_gene_product
and	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
in	O
NSCLC	B-Cancer
and	O
their	O
relations	O
to	O
metastasis	O
and	O
prognosis	O
.	O

The	O
mRNA	O
and	O
protein	O
expressions	O
of	O
KISS1	B-Gene_or_gene_product
and	O
of	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
protein	O
were	O
detected	O
by	O
in	O
situ	O
hybridization	O
and	O
immunohistochemistry	O
respectively	O
in	O
85	O
cases	O
of	O
NSCLC	B-Cancer
,	O
and	O
their	O
matched	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastases	O
.	O

Expressions	O
of	O
KISS1	B-Gene_or_gene_product
mRNA	O
and	O
protein	O
were	O
significantly	O
higher	O
in	O
low	O
TNM	O
stages	O
of	O
NSCLC	B-Cancer
(	O
I	O
-	O
II	O
)	O
compared	O
to	O
more	O
advanced	O
stages	O
(	O
III	O
-	O
IV	O
)	O
(	O
p	O
<	O
0	O
.	O
05	O
)	O
.	O

Moreover	O
,	O
in	O
advanced	O
TNM	O
stages	O
,	O
cases	O
without	O
metastasis	O
had	O
higher	O
KISS1	B-Gene_or_gene_product
gene	O
expression	O
compared	O
to	O
those	O
with	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
(	O
p	O
<	O
0	O
.	O
05	O
)	O
.	O

In	O
contrast	O
,	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
expression	O
was	O
higher	O
in	O
stage	O
III	O
-	O
IV	O
NSCLC	B-Cancer
cases	O
compared	O
to	O
stage	O
I	O
-	O
II	O
tumors	B-Cancer
(	O
p	O
<	O
0	O
.	O
05	O
)	O
and	O
higher	O
in	O
NSCLC	B-Cancer
cases	O
with	O
metastasis	O
than	O
those	O
without	O
metastasis	O
(	O
p	O
<	O
0	O
.	O
05	O
)	O
.	O

There	O
was	O
negative	O
correction	O
between	O
KISS1	B-Gene_or_gene_product
and	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
protein	O
expression	O
(	O
p	O
<	O
0	O
.	O
01	O
)	O
.	O

The	O
5	O
-	O
year	O
survival	O
rate	O
in	O
cases	O
with	O
higher	O
KISS1	B-Gene_or_gene_product
protein	O
expression	O
was	O
significantly	O
higher	O
than	O
in	O
those	O
with	O
low	O
expression	O
(	O
20	O
.	O
9	O
vs	O
.	O
2	O
.	O
4	O
%	O
,	O
p	O
<	O
0	O
.	O
01	O
)	O
.	O

However	O
,	O
the	O
5	O
-	O
year	O
survival	O
rate	O
of	O
patients	B-Organism
with	O
high	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
protein	O
expression	O
were	O
lower	O
than	O
those	O
with	O
low	O
expression	O
(	O
19	O
vs	O
.	O
4	O
.	O
7	O
%	O
,	O
p	O
<	O
0	O
.	O
05	O
)	O
.	O

Our	O
data	O
suggest	O
that	O
KISS1	B-Gene_or_gene_product
and	O
MMP	B-Gene_or_gene_product
-	I-Gene_or_gene_product
9	I-Gene_or_gene_product
may	O
serve	O
as	O
potential	O
prognostic	O
and	O
therapeutic	O
markers	O
in	O
lung	B-Cancer
cancer	I-Cancer
.	O

Relationship	O
and	O
prognostic	O
significance	O
of	O
SPARC	B-Gene_or_gene_product
and	O
VEGF	B-Gene_or_gene_product
protein	O
expression	O
in	O
colon	B-Cancer
cancer	I-Cancer
.	O

BACKGROUND	O
:	O
SPARC	B-Gene_or_gene_product
(	O
secreted	B-Gene_or_gene_product
protein	I-Gene_or_gene_product
,	I-Gene_or_gene_product
acidic	I-Gene_or_gene_product
and	I-Gene_or_gene_product
rich	I-Gene_or_gene_product
in	I-Gene_or_gene_product
cysteine	I-Gene_or_gene_product
)	O
is	O
closely	O
related	O
with	O
the	O
progress	O
,	O
invasion	O
and	O
metastasis	O
of	O
malignant	B-Cancer
tumor	I-Cancer
and	O
angiogenesis	O
.	O

METHODS	O
:	O
Using	O
human	B-Organism
colon	B-Tissue
adenocarcinoma	I-Tissue
tissues	I-Tissue
(	O
hereinafter	O
referred	O
to	O
as	O
colon	B-Cancer
cancer	I-Cancer
)	O
and	O
their	O
corresponding	O
non	B-Tissue
-	I-Tissue
diseased	I-Tissue
colon	I-Tissue
from	O
114	O
patients	B-Organism
'	O
biopsies	B-Tissue
,	O
the	O
expression	O
of	O
SPARC	B-Gene_or_gene_product
and	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
VEGF	B-Gene_or_gene_product
)	O
were	O
investigated	O
by	O
immunohistochemistry	O
staining	O
to	O
assessment	O
the	O
relationship	O
between	O
SPARC	B-Gene_or_gene_product
and	O
VEGF	B-Gene_or_gene_product
,	O
as	O
well	O
as	O
their	O
prognostic	O
significance	O
in	O
patients	B-Organism
.	O

Evaluation	O
of	O
VEGF	B-Gene_or_gene_product
expression	O
level	O
with	O
the	O
same	O
tissues	B-Tissue
was	O
used	O
to	O
establish	O
the	O
antigenic	O
profiles	O
,	O
and	O
the	O
marker	O
of	O
CD34	B-Gene_or_gene_product
staining	O
was	O
used	O
as	O
an	O
indicator	O
of	O
microvessel	B-Tissue
density	O
(	O
MVD	O
)	O
.	O

RESULTS	O
:	O
SPARC	B-Gene_or_gene_product
expression	O
was	O
mainly	O
in	O
the	O
stromal	B-Cell
cells	I-Cell
surrounding	O
the	O
colon	B-Cancer
cancer	I-Cancer
,	O
and	O
was	O
significant	O
difference	O
in	O
those	O
tissues	B-Tissue
with	O
the	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
and	O
differentiation	O
degree	O
of	O
tumor	B-Cancer
.	O

Expression	O
of	O
SPARC	B-Gene_or_gene_product
was	O
significantly	O
correlated	O
with	O
the	O
expression	O
of	O
VEGF	B-Gene_or_gene_product
and	O
MVD	B-Gene_or_gene_product
in	O
colon	B-Tissue
cancer	I-Tissue
tissues	I-Tissue
.	O

Patients	B-Organism
with	O
low	O
or	O
absence	O
expressing	O
SPARC	B-Gene_or_gene_product
had	O
significantly	O
worse	O
overall	O
survival	O
and	O
disease	O
-	O
free	O
survival	O
in	O
a	O
Single	O
Factor	O
Analysis	O
;	O
Cox	O
Regression	O
Analysis	O
,	O
SPARC	B-Gene_or_gene_product
emerged	O
as	O
an	O
overall	O
survival	O
and	O
disease	O
-	O
free	O
survival	O
independent	O
prognostic	O
factor	O
for	O
colon	B-Cancer
cancer	I-Cancer
.	O

CONCLUSION	O
:	O
The	O
low	O
expression	O
or	O
absence	O
of	O
stromal	B-Cell
SPARC	B-Gene_or_gene_product
was	O
an	O
independent	O
prognostic	O
factor	O
for	O
poor	O
prognosis	O
of	O
colon	B-Cancer
cancer	I-Cancer
.	O

SPARC	B-Gene_or_gene_product
maybe	O
involved	O
in	O
the	O
regulation	O
of	O
anti	O
-	O
angiogenesis	O
by	O
which	O
it	O
may	O
serve	O
as	O
a	O
novel	O
target	O
for	O
colon	B-Cancer
cancer	I-Cancer
treatment	O
as	O
well	O
as	O
a	O
novel	O
distinctive	O
marker	O
.	O

The	O
neural	B-Cell
stem	I-Cell
cell	I-Cell
fate	O
determinant	O
TLX	B-Gene_or_gene_product
promotes	O
tumorigenesis	O
and	O
genesis	O
of	O
cells	B-Cell
resembling	O
glioma	B-Cell
stem	I-Cell
cells	I-Cell
.	O

A	O
growing	O
body	B-Organism_subdivision
of	O
evidence	O
indicates	O
that	O
deregulation	O
of	O
stem	B-Cell
cell	I-Cell
fate	O
determinants	O
is	O
a	O
hallmark	O
of	O
many	O
types	O
of	O
malignancies	B-Cancer
.	O

The	O
neural	B-Cell
stem	I-Cell
cell	I-Cell
fate	O
determinant	O
TLX	B-Gene_or_gene_product
plays	O
a	O
pivotal	O
role	O
in	O
neurogenesis	O
in	O
the	O
adult	O
brain	B-Organ
by	O
maintaining	O
neural	B-Cell
stem	I-Cell
cells	I-Cell
.	O

Here	O
,	O
we	O
report	O
a	O
tumorigenic	O
role	O
of	O
TLX	B-Gene_or_gene_product
in	O
brain	B-Cancer
tumor	I-Cancer
initiation	O
and	O
progression	O
.	O

Increased	O
TLX	B-Gene_or_gene_product
expression	O
was	O
observed	O
in	O
a	O
number	O
of	O
glioma	B-Cell
cells	I-Cell
and	O
glioma	B-Cell
stem	I-Cell
cells	I-Cell
,	O
and	O
correlated	O
with	O
poor	O
survival	O
of	O
patients	B-Organism
with	O
gliomas	B-Cancer
.	O

Ectopic	O
expression	O
of	O
TLX	B-Gene_or_gene_product
in	O
the	O
U87MG	B-Cell
glioma	I-Cell
cell	I-Cell
line	I-Cell
and	O
Ink4a	B-Gene_or_gene_product
/	O
Arf	B-Gene_or_gene_product
-	O
deficient	O
mouse	O
astrocytes	O
(	O
Ink4a	B-Gene_or_gene_product
/	O
Arf	B-Gene_or_gene_product
(	O
-	O
/	O
-	O
)	O
astrocytes	O
)	O
induced	O
cell	B-Cell
proliferation	O
with	O
a	O
concomitant	O
increase	O
in	O
cyclin	B-Gene_or_gene_product
D	I-Gene_or_gene_product
expression	O
,	O
and	O
accelerated	O
foci	B-Cell
formation	O
in	O
soft	O
agar	O
and	O
tumor	B-Cancer
formation	O
in	O
in	O
vivo	O
transplantation	O
assays	O
.	O

Furthermore	O
,	O
overexpression	O
of	O
TLX	B-Gene_or_gene_product
in	O
Ink4a	B-Gene_or_gene_product
/	O
Arf	B-Gene_or_gene_product
(	O
-	O
/	O
-	O
)	O
astrocytes	O
inhibited	O
cell	B-Cell
migration	O
and	O
invasion	O
and	O
promoted	O
neurosphere	B-Cell
formation	O
and	O
Nestin	B-Gene_or_gene_product
expression	O
,	O
which	O
are	O
hallmark	O
characteristics	O
of	O
glioma	B-Cell
stem	I-Cell
cells	I-Cell
,	O
under	O
stem	B-Cell
cell	I-Cell
culture	O
conditions	O
.	O

Our	O
results	O
indicate	O
that	O
TLX	B-Gene_or_gene_product
is	O
involved	O
in	O
glioma	B-Cell
stem	I-Cell
cell	I-Cell
genesis	O
and	O
represents	O
a	O
potential	O
therapeutic	O
target	O
for	O
this	O
type	O
of	O
malignancy	B-Cancer
.	O

Expression	O
profiling	O
in	O
progressive	O
stages	O
of	O
fumarate	B-Gene_or_gene_product
-	I-Gene_or_gene_product
hydratase	I-Gene_or_gene_product
deficiency	O
:	O
the	O
contribution	O
of	O
metabolic	O
changes	O
to	O
tumorigenesis	O
.	O

Hereditary	B-Cancer
leiomyomatosis	I-Cancer
and	I-Cancer
renal	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
(	O
HLRCC	B-Cancer
)	O
is	O
caused	O
by	O
mutations	O
in	O
the	O
Krebs	O
cycle	O
enzyme	O
fumarate	B-Gene_or_gene_product
hydratase	I-Gene_or_gene_product
(	O
FH	B-Gene_or_gene_product
)	O
.	O

It	O
has	O
been	O
proposed	O
that	O
"	O
pseudohypoxic	O
"	O
stabilization	O
of	O
hypoxia	B-Gene_or_gene_product
-	I-Gene_or_gene_product
inducible	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
(	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
)	O
by	O
fumarate	B-Simple_chemical
accumulation	O
contributes	O
to	O
tumorigenesis	O
in	O
HLRCC	B-Cancer
.	O

We	O
hypothesized	O
that	O
an	O
additional	O
direct	O
consequence	O
of	O
FH	B-Gene_or_gene_product
deficiency	O
is	O
the	O
establishment	O
of	O
a	O
biosynthetic	O
milieu	O
.	O

To	O
investigate	O
this	O
hypothesis	O
,	O
we	O
isolated	O
primary	B-Cell
mouse	I-Cell
embryonic	I-Cell
fibroblast	I-Cell
(	I-Cell
MEF	I-Cell
)	I-Cell
lines	I-Cell
from	O
Fh1	B-Gene_or_gene_product
-	O
deficient	O
mice	O
.	O

As	O
predicted	O
,	O
these	O
MEFs	B-Cell
upregulated	O
Hif	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
and	O
HIF	B-Gene_or_gene_product
target	O
genes	O
directly	O
as	O
a	O
result	O
of	O
FH	B-Gene_or_gene_product
deficiency	O
.	O

In	O
addition	O
,	O
detailed	O
metabolic	O
assessment	O
of	O
these	O
MEFs	B-Cell
confirmed	O
their	O
dependence	O
on	O
glycolysis	O
,	O
and	O
an	O
elevated	O
rate	O
of	O
lactate	B-Simple_chemical
efflux	O
,	O
associated	O
with	O
the	O
upregulation	O
of	O
glycolytic	B-Gene_or_gene_product
enzymes	I-Gene_or_gene_product
known	O
to	O
be	O
associated	O
with	O
tumorigenesis	O
.	O

Correspondingly	O
,	O
Fh1	B-Gene_or_gene_product
-	O
deficient	O
benign	O
murine	O
renal	O
cysts	O
and	O
an	O
advanced	O
human	B-Organism
HLRCC	B-Cancer
-	I-Cancer
related	I-Cancer
renal	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
manifested	O
a	O
prominent	O
and	O
progressive	O
increase	O
in	O
the	O
expression	O
of	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
target	O
genes	O
and	O
in	O
genes	O
known	O
to	O
be	O
relevant	O
to	O
tumorigenesis	O
and	O
metastasis	O
.	O

In	O
accord	O
with	O
our	O
hypothesis	O
,	O
in	O
a	O
variety	O
of	O
different	O
FH	B-Gene_or_gene_product
-	O
deficient	O
tissues	O
,	O
including	O
a	O
novel	O
murine	B-Organism
model	O
of	O
Fh1	B-Gene_or_gene_product
-	O
deficient	O
smooth	O
muscle	O
,	O
we	O
show	O
a	O
striking	O
and	O
progressive	O
upregulation	O
of	O
a	O
tumorigenic	O
metabolic	O
profile	O
,	O
as	O
manifested	O
by	O
increased	O
PKM2	B-Gene_or_gene_product
and	O
LDHA	B-Gene_or_gene_product
protein	O
.	O

Based	O
on	O
the	O
models	O
assessed	O
herein	B-Simple_chemical
,	O
we	O
infer	O
that	O
that	O
FH	B-Gene_or_gene_product
deficiency	O
compels	O
cells	B-Cell
to	O
adopt	O
an	O
early	O
,	O
reversible	O
,	O
and	O
progressive	O
protumorigenic	O
metabolic	O
milieu	O
that	O
is	O
reminiscent	O
of	O
that	O
driving	O
the	O
Warburg	O
effect	O
.	O

Targets	O
identified	O
in	O
these	O
novel	O
and	O
diverse	O
FH	B-Gene_or_gene_product
-	O
deficient	O
models	O
represent	O
excellent	O
potential	O
candidates	O
for	O
further	O
mechanistic	O
investigation	O
and	O
therapeutic	O
metabolic	O
manipulation	O
in	O
tumors	B-Cancer
.	O

A	O
STAT3	B-Gene_or_gene_product
-	O
mediated	O
metabolic	O
switch	O
is	O
involved	O
in	O
tumour	B-Cancer
transformation	O
and	O
STAT3	B-Gene_or_gene_product
addiction	O
.	O

The	O
pro	O
-	O
oncogenic	O
transcription	O
factor	O
STAT3	B-Gene_or_gene_product
is	O
constitutively	O
activated	O
in	O
a	O
wide	O
variety	O
of	O
tumours	B-Cancer
that	O
often	O
become	O
addicted	O
to	O
its	O
activity	O
,	O
but	O
no	O
unifying	O
view	O
of	O
a	O
core	O
function	O
determining	O
this	O
widespread	O
STAT3	B-Gene_or_gene_product
-	O
dependence	O
has	O
yet	O
emerged	O
.	O

We	O
show	O
here	O
that	O
constitutively	O
active	O
STAT3	B-Gene_or_gene_product
acts	O
as	O
a	O
master	O
regulator	O
of	O
cell	B-Cell
metabolism	O
,	O
inducing	O
aerobic	O
glycolysis	O
and	O
down	O
-	O
regulating	O
mitochondrial	B-Cellular_component
activity	O
both	O
in	O
primary	O
fibroblasts	B-Cell
and	O
in	O
STAT3	B-Gene_or_gene_product
-	O
dependent	O
tumour	B-Cell
cell	I-Cell
lines	I-Cell
.	O

As	O
a	O
result	O
,	O
cells	B-Cell
are	O
protected	O
from	O
apoptosis	O
and	O
senescence	O
while	O
becoming	O
highly	O
sensitive	O
to	O
glucose	B-Simple_chemical
deprivation	O
.	O

We	O
show	O
that	O
enhanced	O
glycolysis	O
is	O
dependent	O
on	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
up	O
-	O
regulation	O
,	O
while	O
reduced	O
mitochondrial	B-Cellular_component
activity	O
is	O
HIF	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1alpha	I-Gene_or_gene_product
-	O
independent	O
and	O
likely	O
caused	O
by	O
STAT3	B-Gene_or_gene_product
-	O
mediated	O
down	O
-	O
regulation	O
of	O
mitochondrial	B-Cellular_component
proteins	O
.	O

The	O
induction	O
of	O
aerobic	O
glycolysis	O
is	O
an	O
important	O
component	O
of	O
STAT3	B-Gene_or_gene_product
pro	O
-	O
oncogenic	O
activities	O
,	O
since	O
inhibition	O
of	O
STAT3	B-Gene_or_gene_product
tyrosine	B-Amino_acid
phosphorylation	O
in	O
the	O
tumour	B-Cell
cell	I-Cell
lines	I-Cell
down	O
-	O
regulates	O
glycolysis	O
prior	O
to	O
leading	O
to	O
growth	O
arrest	O
and	O
cell	B-Cell
death	O
,	O
both	O
in	O
vitro	O
and	O
in	O
vivo	O
.	O

We	O
propose	O
that	O
this	O
novel	O
,	O
central	O
metabolic	O
role	O
is	O
at	O
the	O
core	O
of	O
the	O
addiction	O
for	O
STAT3	B-Gene_or_gene_product
shown	O
by	O
so	O
many	O
biologically	O
different	O
tumours	B-Cancer
.	O

Roles	O
of	O
brca2	B-Gene_or_gene_product
(	O
fancd1	B-Gene_or_gene_product
)	O
in	O
oocyte	B-Cellular_component
nuclear	I-Cellular_component
architecture	O
,	O
gametogenesis	O
,	O
gonad	B-Cancer
tumors	I-Cancer
,	O
and	O
genome	O
stability	O
in	O
zebrafish	B-Organism
.	O

Mild	O
mutations	O
in	O
BRCA2	B-Gene_or_gene_product
(	O
FANCD1	B-Gene_or_gene_product
)	O
cause	O
Fanconi	O
anemia	O
(	O
FA	O
)	O
when	O
homozygous	O
,	O
while	O
severe	O
mutations	O
cause	O
common	O
cancers	B-Cancer
including	O
breast	B-Cancer
,	O
ovarian	B-Cancer
,	O
and	O
prostate	B-Cancer
cancers	I-Cancer
when	O
heterozygous	O
.	O

Here	O
we	O
report	O
a	O
zebrafish	O
brca2	B-Gene_or_gene_product
insertional	O
mutant	O
that	O
shares	O
phenotypes	O
with	O
human	B-Organism
patients	I-Organism
and	O
identifies	O
a	O
novel	O
brca2	B-Gene_or_gene_product
function	O
in	O
oogenesis	O
.	O

Experiments	O
showed	O
that	O
mutant	B-Developing_anatomical_structure
embryos	I-Developing_anatomical_structure
and	O
mutant	B-Cell
cells	I-Cell
in	O
culture	O
experienced	O
genome	O
instability	O
,	O
as	O
do	O
cells	B-Cell
in	O
FA	O
patients	B-Organism
.	O

In	O
wild	O
-	O
type	O
zebrafish	B-Organism
,	O
meiotic	O
cells	B-Cell
expressed	O
brca2	B-Gene_or_gene_product
;	O
and	O
,	O
unexpectedly	O
,	O
transcripts	O
in	O
oocytes	B-Cell
localized	O
asymmetrically	O
to	O
the	O
animal	B-Cellular_component
pole	I-Cellular_component
.	O

In	O
juvenile	O
brca2	B-Gene_or_gene_product
mutants	O
,	O
oocytes	B-Cell
failed	O
to	O
progress	O
through	O
meiosis	O
,	O
leading	O
to	O
female	O
-	O
to	O
-	O
male	O
sex	O
reversal	O
.	O

Adult	B-Organism
mutants	I-Organism
became	O
sterile	O
males	O
due	O
to	O
the	O
meiotic	O
arrest	O
of	O
spermatocytes	B-Cell
,	O
which	O
then	O
died	O
by	O
apoptosis	O
,	O
followed	O
by	O
neoplastic	B-Cancer
proliferation	O
of	O
gonad	B-Cell
somatic	I-Cell
cells	I-Cell
that	O
was	O
similar	O
to	O
neoplasia	B-Pathological_formation
observed	O
in	O
ageing	O
dead	B-Gene_or_gene_product
end	I-Gene_or_gene_product
(	O
dnd	B-Gene_or_gene_product
)	O
-	O
knockdown	O
males	O
,	O
which	O
lack	O
germ	B-Cell
cells	I-Cell
.	O

The	O
construction	O
of	O
animals	B-Organism
doubly	I-Organism
mutant	I-Organism
for	O
brca2	B-Gene_or_gene_product
and	O
the	O
apoptotic	O
gene	O
tp53	B-Gene_or_gene_product
(	O
p53	B-Gene_or_gene_product
)	O
rescued	O
brca2	B-Gene_or_gene_product
-	O
dependent	O
sex	O
reversal	O
.	O

Double	B-Organism
mutants	I-Organism
developed	O
oocytes	B-Cell
and	O
became	O
sterile	O
females	O
that	O
produced	O
only	O
aberrant	O
embryos	B-Developing_anatomical_structure
and	O
showed	O
elevated	O
risk	O
for	O
invasive	O
ovarian	B-Cancer
tumors	I-Cancer
.	O

Oocytes	B-Cell
in	O
double	B-Organism
-	I-Organism
mutant	I-Organism
females	I-Organism
showed	O
normal	O
localization	O
of	O
brca2	B-Gene_or_gene_product
and	O
pou5f1	B-Gene_or_gene_product
transcripts	O
to	O
the	O
animal	B-Cellular_component
pole	I-Cellular_component
and	O
vasa	O
transcripts	O
to	O
the	O
vegetal	B-Cellular_component
pole	I-Cellular_component
,	O
but	O
had	O
a	O
polarized	O
rather	O
than	O
symmetrical	O
nucleus	B-Cellular_component
with	O
the	O
distribution	O
of	O
nucleoli	B-Cellular_component
and	O
chromosomes	B-Cellular_component
to	O
opposite	O
nuclear	B-Cellular_component
poles	I-Cellular_component
;	O
this	O
result	O
revealed	O
a	O
novel	O
role	O
for	O
Brca2	B-Gene_or_gene_product
in	O
establishing	O
or	O
maintaining	O
oocyte	B-Cell
nuclear	B-Cellular_component
architecture	O
.	O

Mutating	O
tp53	B-Gene_or_gene_product
did	O
not	O
rescue	O
the	O
infertility	O
phenotype	O
in	O
brca2	B-Gene_or_gene_product
mutant	O
males	O
,	O
suggesting	O
that	O
brca2	B-Gene_or_gene_product
plays	O
an	O
essential	O
role	O
in	O
zebrafish	B-Organism
spermatogenesis	O
.	O

Overall	O
,	O
this	O
work	O
verified	O
zebrafish	B-Organism
as	O
a	O
model	O
for	O
the	O
role	O
of	O
Brca2	B-Gene_or_gene_product
in	O
human	B-Organism
disease	O
and	O
uncovered	O
a	O
novel	O
function	O
of	O
Brca2	B-Gene_or_gene_product
in	O
vertebrate	O
oocyte	B-Cellular_component
nuclear	I-Cellular_component
architecture	O
.	O

KISS1	B-Gene_or_gene_product
methylation	O
and	O
expression	O
as	O
tumor	B-Cancer
stratification	O
biomarkers	O
and	O
clinical	O
outcome	O
prognosticators	O
for	O
bladder	B-Cancer
cancer	I-Cancer
patients	B-Organism
.	O

KISS1	B-Gene_or_gene_product
is	O
a	O
metastasis	O
suppressor	O
gene	O
that	O
is	O
lost	O
in	O
several	O
malignancies	B-Cancer
,	O
including	O
bladder	B-Cancer
cancer	I-Cancer
.	O

We	O
tested	O
the	O
epigenetic	O
silencing	O
hypothesis	O
and	O
evaluated	O
the	O
biological	O
influence	O
of	O
KISS1	B-Gene_or_gene_product
methylation	O
on	O
its	O
expression	O
and	O
clinical	O
relevance	O
in	O
bladder	B-Cancer
cancer	I-Cancer
.	O

KISS1	B-Gene_or_gene_product
hypermethylation	O
was	O
frequent	O
in	O
bladder	B-Cell
cancer	I-Cell
cells	I-Cell
analyzed	O
by	O
methylation	O
-	O
specific	O
PCR	O
and	O
bisulfite	B-Simple_chemical
sequencing	O
and	O
was	O
associated	O
with	O
low	O
gene	O
expression	O
,	O
being	O
restored	O
in	O
vitro	O
by	O
demethylating	O
azacytidine	B-Simple_chemical
.	O

Hypermethylation	O
was	O
also	O
frequently	O
observed	O
in	O
a	O
large	O
series	O
of	O
bladder	B-Cancer
tumors	I-Cancer
(	O
83	O
.	O
1	O
%	O
,	O
n	O
=	O
804	O
)	O
.	O

KISS1	B-Gene_or_gene_product
methylation	O
was	O
associated	O
with	O
increasing	O
stage	O
(	O
P	O
=	O
0	O
.	O
001	O
)	O
and	O
tumor	B-Cancer
grade	O
(	O
P	O
=	O
0	O
.	O
010	O
)	O
.	O

KISS1	B-Gene_or_gene_product
methylation	O
was	O
associated	O
with	O
low	O
KISS1	B-Gene_or_gene_product
transcript	O
expression	O
by	O
quantitative	O
RT	O
-	O
PCR	O
(	O
P	O
=	O
0	O
.	O
037	O
)	O
.	O

KISS1	B-Gene_or_gene_product
transcript	O
expression	O
was	O
also	O
associated	O
with	O
histopathological	O
tumor	B-Cancer
stage	O
(	O
P	O
<	O
0	O
.	O
0005	O
)	O
.	O

Low	O
transcript	O
expression	O
alone	O
(	O
P	O
=	O
0	O
.	O
003	O
)	O
or	O
combined	O
with	O
methylation	O
(	O
P	O
=	O
0	O
.	O
019	O
)	O
was	O
associated	O
with	O
poor	O
disease	O
-	O
specific	O
survival	O
(	O
n	O
=	O
205	O
)	O
.	O

KISS1	B-Gene_or_gene_product
transcript	O
expression	O
remained	O
an	O
independent	O
prognosticator	O
in	O
multivariate	O
analyses	O
(	O
P	O
=	O
0	O
.	O
017	O
)	O
.	O

KISS1	B-Gene_or_gene_product
hypermethylation	O
was	O
identified	O
in	O
bladder	B-Cancer
cancer	I-Cancer
,	O
providing	O
a	O
potential	O
mechanistic	O
explanation	O
(	O
epigenetic	O
silencing	O
)	O
for	O
the	O
observed	O
loss	O
of	O
KISS1	B-Gene_or_gene_product
in	O
uroepithelial	B-Cancer
malignancies	I-Cancer
.	O

Associations	O
of	O
KISS1	B-Gene_or_gene_product
methylation	O
and	O
its	O
expression	O
with	O
histopathological	O
variables	O
and	O
poor	O
survival	O
suggest	O
the	O
utility	O
of	O
incorporating	O
KISS1	B-Gene_or_gene_product
measurement	O
using	O
paraffin	B-Simple_chemical
-	O
embedded	O
material	O
for	O
tumor	B-Cancer
stratification	O
and	O
clinical	O
outcome	O
prognosis	O
of	O
patients	B-Organism
with	O
uroepithelial	B-Cancer
neoplasias	I-Cancer
.	O

Correlation	O
of	O
lymphatic	B-Multi-tissue_structure
vessel	I-Multi-tissue_structure
density	O
and	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
with	O
nodal	B-Multi-tissue_structure
metastasis	O
in	O
papillary	B-Cancer
thyroid	I-Cancer
microcarcinoma	I-Cancer
.	O

BACKGROUND	O
:	O
The	O
aim	O
of	O
this	O
study	O
was	O
to	O
investigate	O
not	O
only	O
the	O
intratumoral	B-Cancer
and	O
peritumoral	B-Multi-tissue_structure
lymphatic	I-Multi-tissue_structure
vessel	I-Multi-tissue_structure
density	O
(	O
ILD	O
and	O
PLD	O
)	O
but	O
also	O
the	O
expression	O
of	O
vascular	B-Gene_or_gene_product
endothelial	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factors	I-Gene_or_gene_product
(	O
VEGFs	B-Gene_or_gene_product
)	O
and	O
to	O
test	O
their	O
correlation	O
with	O
regional	O
lymph	B-Multi-tissue_structure
nodal	I-Multi-tissue_structure
metastases	O
in	O
papillary	B-Cancer
thyroid	I-Cancer
microcarcinoma	I-Cancer
(	O
PTMC	B-Cancer
)	O
.	O

METHODS	O
:	O
A	O
clinicopathologic	O
data	O
review	O
was	O
performed	O
in	O
60	O
patients	B-Organism
with	O
PTMC	B-Cancer
treated	O
with	O
total	O
thyroidectomies	O
involving	O
neck	B-Organism_subdivision
dissections	O
.	O

The	O
patterns	O
of	O
lymphatic	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
,	O
the	O
expression	O
of	O
VEGFs	B-Gene_or_gene_product
,	O
and	O
their	O
correlation	O
with	O
neck	B-Organism_subdivision
metastases	O
were	O
assessed	O
.	O

RESULTS	O
:	O
PLD	O
was	O
significantly	O
higher	O
than	O
ILD	O
(	O
p	O
<	O
.	O
001	O
)	O
.	O

Patients	B-Organism
with	O
high	O
PLD	O
(	O
>	O
8	O
.	O
0	O
)	O
showed	O
higher	O
rates	O
of	O
neck	B-Organism_subdivision
metastases	O
than	O
low	O
PLD	O
(	O
<	O
=	O
8	O
.	O
0	O
)	O
(	O
74	O
.	O
0	O
%	O
vs	O
46	O
.	O
8	O
%	O
,	O
p	O
=	O
.	O
03	O
)	O
.	O

Univariate	O
and	O
multivariate	O
analyses	O
revealed	O
high	O
PLD	O
as	O
the	O
only	O
independent	O
variable	O
predictive	O
of	O
neck	B-Organism_subdivision
metastasis	O
.	O

The	O
expression	O
of	O
VEGFs	B-Gene_or_gene_product
did	O
not	O
correlate	O
with	O
either	O
lymphatic	B-Organ
density	O
or	O
neck	B-Organism_subdivision
metastasis	O
.	O

CONCLUSIONS	O
:	O
PLD	O
may	O
be	O
of	O
potential	O
benefit	O
in	O
the	O
prediction	O
of	O
neck	B-Organism_subdivision
metastasis	O
in	O
PTMC	B-Cancer
.	O

Inhibition	O
of	O
autophagy	O
potentiates	O
the	O
antitumor	B-Cancer
effect	O
of	O
the	O
multikinase	O
inhibitor	O
sorafenib	B-Simple_chemical
in	O
hepatocellular	B-Cancer
carcinoma	I-Cancer
.	O

Multikinase	O
inhibitor	O
sorafenib	B-Simple_chemical
inhibits	O
proliferation	O
and	O
angiogenesis	O
of	O
tumors	B-Cancer
by	O
suppressing	O
the	O
Raf	B-Gene_or_gene_product
/	O
MEK	B-Gene_or_gene_product
/	O
ERK	B-Gene_or_gene_product
signaling	O
pathway	O
and	O
VEGF	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
tyrosine	B-Amino_acid
kinase	O
.	O

It	O
significantly	O
prolongs	O
median	O
survival	O
of	O
patients	B-Organism
with	O
advanced	O
hepatocellular	B-Cancer
carcinoma	I-Cancer
(	O
HCC	B-Cancer
)	O
but	O
the	O
response	O
is	O
disease	O
-	O
stabilizing	O
and	O
cytostatic	O
rather	O
than	O
one	O
of	O
tumor	B-Cancer
regression	O
.	O

To	O
examine	O
the	O
mechanisms	O
underlying	O
the	O
relative	O
resistance	O
in	O
HCC	B-Cancer
,	O
we	O
investigated	O
the	O
role	O
of	O
autophagy	O
,	O
an	O
evolutionarily	O
conserved	O
self	O
-	O
digestion	O
pathway	O
,	O
in	O
hepatoma	B-Cell
cells	I-Cell
in	O
vitro	O
and	O
in	O
vivo	O
.	O

Sorafenib	B-Simple_chemical
treatment	O
led	O
to	O
accumulation	O
of	O
autophagosomes	B-Cellular_component
as	O
evidenced	O
by	O
conversion	O
from	O
LC3	B-Gene_or_gene_product
-	I-Gene_or_gene_product
I	I-Gene_or_gene_product
to	O
LC3	B-Gene_or_gene_product
-	I-Gene_or_gene_product
II	I-Gene_or_gene_product
observed	O
by	O
immunoblot	O
in	O
Huh7	B-Cell
,	O
HLF	B-Cell
and	O
PLC	B-Cell
/	I-Cell
PRF	I-Cell
/	I-Cell
5	I-Cell
cells	I-Cell
.	O

This	O
induction	O
was	O
due	O
to	O
activation	O
of	O
autophagic	O
flux	O
,	O
as	O
there	O
was	O
further	O
increase	O
in	O
LC3	B-Gene_or_gene_product
-	I-Gene_or_gene_product
II	I-Gene_or_gene_product
expression	O
upon	O
treatment	O
with	O
lysosomal	O
inhibitors	O
,	O
clear	O
decline	O
of	O
the	O
autophagy	O
substrate	O
p62	B-Gene_or_gene_product
,	O
and	O
an	O
mRFP	B-Gene_or_gene_product
-	O
GFP	B-Gene_or_gene_product
-	O
LC3	B-Gene_or_gene_product
fluorescence	O
change	O
in	O
sorafenib	B-Simple_chemical
-	O
treated	O
hepatoma	B-Cell
cells	I-Cell
.	O

Sorafenib	B-Simple_chemical
inhibited	O
the	O
mammalian	B-Gene_or_gene_product
target	I-Gene_or_gene_product
of	I-Gene_or_gene_product
rapamycin	I-Gene_or_gene_product
complex	I-Gene_or_gene_product
1	I-Gene_or_gene_product
and	O
its	O
inhibition	O
led	O
to	O
accumulation	O
of	O
LC3	B-Gene_or_gene_product
-	I-Gene_or_gene_product
II	I-Gene_or_gene_product
.	O

Pharmacological	O
inhibition	O
of	O
autophagic	O
flux	O
by	O
chloroquine	B-Simple_chemical
increased	O
apoptosis	O
and	O
decreased	O
cell	B-Cell
viability	O
in	O
hepatoma	B-Cell
cells	I-Cell
.	O

siRNA	O
-	O
mediated	O
knockdown	O
of	O
the	O
ATG7	B-Gene_or_gene_product
gene	O
also	O
sensitized	O
hepatoma	B-Cell
cells	I-Cell
to	O
sorafenib	B-Simple_chemical
.	O

Finally	O
,	O
sorafenib	B-Simple_chemical
induced	O
autophagy	O
in	O
Huh7	B-Cancer
xenograft	I-Cancer
tumors	I-Cancer
in	O
nude	B-Organism
mice	I-Organism
and	O
coadministration	O
with	O
chloroquine	B-Simple_chemical
significantly	O
suppressed	O
tumor	B-Cancer
growth	O
compared	O
with	O
sorafenib	B-Simple_chemical
alone	O
.	O

In	O
conclusion	O
,	O
sorafenib	B-Simple_chemical
administration	O
induced	O
autophagosome	B-Cellular_component
formation	O
and	O
enhanced	O
autophagic	O
activity	O
,	O
which	O
conferred	O
a	O
survival	O
advantage	O
to	O
hepatoma	B-Cell
cells	I-Cell
.	O

Concomitant	O
inhibition	O
of	O
autophagy	O
may	O
be	O
an	O
attractive	O
strategy	O
for	O
unlocking	O
the	O
antitumor	B-Cancer
potential	O
of	O
sorafenib	B-Simple_chemical
in	O
HCC	B-Cancer
.	O

Desensitization	O
of	O
prostaglandin	B-Simple_chemical
F2	I-Simple_chemical
alpha	I-Simple_chemical
-	O
stimulated	O
inositol	B-Simple_chemical
phosphate	I-Simple_chemical
generation	O
in	O
NIH	B-Cell
-	I-Cell
3T3	I-Cell
fibroblasts	I-Cell
transformed	O
by	O
overexpression	O
of	O
normal	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Ha	I-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
,	O
c	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
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
N	I-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
genes	O
.	O

The	O
stimulation	O
of	O
inositol	B-Simple_chemical
phosphate	I-Simple_chemical
generation	O
in	O
control	B-Cell
and	O
ras	B-Gene_or_gene_product
-	O
gene	O
-	O
transformed	O
NIH	B-Cell
-	I-Cell
3T3	I-Cell
cells	I-Cell
by	O
prostaglandin	B-Simple_chemical
F2	I-Simple_chemical
alpha	I-Simple_chemical
(	O
PGF2	B-Simple_chemical
alpha	I-Simple_chemical
)	O
was	O
investigated	O
.	O

Compared	O
with	O
the	O
control	B-Cell
cells	I-Cell
,	O
a	O
desensitization	O
of	O
the	O
response	O
was	O
observed	O
in	O
cells	B-Cell
transformed	O
by	O
the	O
overexpression	O
of	O
N	B-Gene_or_gene_product
-	I-Gene_or_gene_product
,	O
Ha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
,	O
or	O
Ki	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
genes	O
.	O

This	O
desensitization	O
was	O
without	O
effect	O
upon	O
the	O
concentration	O
causing	O
half	O
-	O
maximal	O
effect	O
(	O
EC50	O
)	O
,	O
dissociation	O
constant	O
(	O
Kd	O
)	O
or	O
number	O
of	O
PGF2	B-Gene_or_gene_product
alpha	I-Gene_or_gene_product
receptors	I-Gene_or_gene_product
.	O

Inhibition	O
of	O
PG	B-Simple_chemical
synthesis	O
was	O
without	O
effect	O
upon	O
desensitization	O
,	O
demonstrating	O
that	O
the	O
effect	O
was	O
not	O
agonist	O
-	O
induced	O
.	O

Desensitization	O
could	O
be	O
induced	O
in	O
NIH	B-Cell
-	I-Cell
3T3	I-Cell
cells	I-Cell
by	O
culturing	O
under	O
conditions	O
where	O
the	O
cells	B-Cell
were	O
all	O
in	O
the	O
exponential	O
growth	O
phase	O
,	O
or	O
by	O
a	O
12	O
h	O
exposure	O
to	O
a	O
C	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
-	O
activating	O
phorbol	B-Simple_chemical
ester	I-Simple_chemical
.	O

These	O
results	O
suggest	O
that	O
desensitization	O
of	O
certain	O
agonist	O
-	O
induced	O
inositol	B-Simple_chemical
phospholipid	I-Simple_chemical
responses	O
in	O
ras	B-Gene_or_gene_product
-	O
transformed	O
cells	B-Cell
is	O
a	O
consequence	O
of	O
increased	O
cell	B-Cell
proliferation	O
and	O
associated	O
amplification	O
in	O
C	B-Gene_or_gene_product
-	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
activity	O
and	O
is	O
an	O
indirect	O
consequence	O
of	O
transformation	O
by	O
ras	B-Gene_or_gene_product
.	O

Relationship	O
between	O
of	O
blood	B-Organism_substance
flow	O
,	O
glucose	B-Simple_chemical
metabolism	O
,	O
protein	O
synthesis	O
,	O
glucose	B-Simple_chemical
and	O
ATP	B-Simple_chemical
content	O
in	O
experimentally	O
-	O
induced	O
glioma	B-Cancer
(	O
RG1	B-Cancer
2	I-Cancer
.	I-Cancer
2	I-Cancer
)	O
of	O
rat	B-Organism
brain	B-Organ
.	O

In	O
experimental	O
RG1	B-Cancer
2	I-Cancer
.	I-Cancer
2	I-Cancer
glioma	I-Cancer
of	O
rat	B-Organism
brain	B-Organ
,	O
local	O
blood	B-Organism_substance
flow	O
,	O
glucose	B-Simple_chemical
utilization	O
,	O
protein	O
synthesis	O
,	O
glucose	B-Simple_chemical
and	O
ATP	B-Simple_chemical
content	O
were	O
measured	O
by	O
means	O
of	O
triple	O
tracer	O
autoradiography	O
and	O
bioluminescence	O
technique	O
,	O
respectively	O
,	O
to	O
determine	O
hemodynamic	O
and	O
metabolic	O
thresholds	O
for	O
local	O
tumor	B-Cancer
energy	O
failure	O
.	O

Perfusion	O
thresholds	O
were	O
estimated	O
at	O
tumor	B-Cancer
blood	B-Organism_substance
flow	O
values	O
of	O
69	O
.	O
0	O
+	O
/	O
-	O
0	O
.	O
1	O
ml	O
/	O
100	O
g	O
/	O
min	O
(	O
estimate	O
+	O
/	O
-	O
standard	O
error	O
)	O
and	O
of	O
69	O
+	O
/	O
-	O
7	O
.	O
1	O
ml	O
/	O
100	O
g	O
/	O
min	O
for	O
the	O
beginning	O
of	O
the	O
decline	O
in	O
regional	O
ATP	B-Simple_chemical
and	O
glucose	B-Simple_chemical
content	O
,	O
respectively	O
.	O

Metabolic	O
thresholds	O
were	O
derived	O
at	O
tumor	B-Cancer
glucose	B-Simple_chemical
utilization	O
values	O
of	O
70	O
.	O
6	O
+	O
/	O
-	O
8	O
.	O
3	O
mumol	O
/	O
100	O
g	O
/	O
min	O
for	O
reduced	O
protein	O
synthesis	O
,	O
of	O
55	O
.	O
0	O
+	O
/	O
-	O
0	O
.	O
2	O
mumol	O
/	O
100	O
g	O
/	O
min	O
for	O
the	O
decrease	O
in	O
glucose	B-Simple_chemical
content	O
,	O
and	O
34	O
.	O
7	O
+	O
/	O
-	O
4	O
.	O
7	O
mumol	O
/	O
100	O
g	O
/	O
min	O
for	O
decline	O
in	O
ATP	B-Simple_chemical
content	O
.	O

Our	O
results	O
suggest	O
that	O
blood	B-Organism_substance
flow	O
limits	O
glucose	B-Simple_chemical
supply	O
to	O
tumor	B-Tissue
tissue	I-Tissue
at	O
much	O
higher	O
flow	O
rates	O
than	O
in	O
normal	O
brain	B-Organ
which	O
,	O
in	O
turn	O
,	O
is	O
associated	O
with	O
a	O
decrease	O
in	O
tumor	B-Cancer
glucose	B-Simple_chemical
utilization	O
.	O

A	O
reduction	O
and	O
not	O
an	O
increase	O
in	O
tumor	B-Cancer
glucose	B-Simple_chemical
availability	O
could	O
be	O
a	O
more	O
appropriate	O
strategy	O
for	O
the	O
induction	O
of	O
energy	O
failure	O
in	O
tumors	B-Cancer
.	O

Inactivation	O
of	O
androgen	B-Simple_chemical
-	O
induced	O
regulator	O
ARD1	B-Gene_or_gene_product
inhibits	O
androgen	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
acetylation	O
and	O
prostate	B-Organ
tumorigenesis	O
.	O

Androgen	B-Simple_chemical
signaling	O
through	O
androgen	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
(	O
AR	B-Gene_or_gene_product
)	O
is	O
critical	O
for	O
prostate	B-Organ
tumorigenesis	O
.	O

Given	O
that	O
AR	B-Gene_or_gene_product
-	O
mediated	O
gene	O
regulation	O
is	O
enhanced	O
by	O
AR	B-Gene_or_gene_product
coregulators	O
,	O
inactivation	O
of	O
those	O
coregulators	O
is	O
emerging	O
as	O
a	O
promising	O
therapy	O
for	O
prostate	B-Cancer
cancer	I-Cancer
(	O
PCa	B-Cancer
)	O
.	O

Here	O
,	O
we	O
show	O
that	O
the	O
N	B-Gene_or_gene_product
-	I-Gene_or_gene_product
acetyltransferase	I-Gene_or_gene_product
arrest	I-Gene_or_gene_product
-	I-Gene_or_gene_product
defect	I-Gene_or_gene_product
1	I-Gene_or_gene_product
protein	O
(	O
ARD1	B-Gene_or_gene_product
)	O
functions	O
as	O
a	O
unique	O
AR	B-Gene_or_gene_product
regulator	O
in	O
PCa	B-Cell
cells	I-Cell
.	O

ARD1	B-Gene_or_gene_product
is	O
up	O
-	O
regulated	O
in	O
human	B-Organism
PCa	B-Cell
cell	I-Cell
lines	I-Cell
and	O
primary	O
tumor	B-Cancer
biopsies	I-Cancer
.	O

The	O
expression	O
of	O
ARD1	B-Gene_or_gene_product
was	O
augmented	O
by	O
treatment	O
with	O
synthetic	O
androgen	B-Simple_chemical
(	O
R1881	B-Simple_chemical
)	O
unless	O
AR	B-Gene_or_gene_product
is	O
deficient	O
or	O
is	O
inhibited	O
by	O
AR	B-Gene_or_gene_product
-	O
specific	O
siRNA	O
or	O
androgen	B-Simple_chemical
inhibitor	O
bicalutamide	B-Simple_chemical
(	O
Casodex	B-Simple_chemical
)	O
.	O

Depletion	O
of	O
ARD1	B-Gene_or_gene_product
by	O
shRNA	O
suppressed	O
PCa	B-Cell
cell	I-Cell
proliferation	O
,	O
anchorage	O
-	O
independent	O
growth	O
,	O
and	O
xenograft	B-Cancer
tumor	I-Cancer
formation	O
in	O
SCID	B-Organism
mice	I-Organism
,	O
suggesting	O
that	O
AR	B-Gene_or_gene_product
-	O
dependent	O
ARD1	B-Gene_or_gene_product
expression	O
is	O
biologically	O
germane	O
.	O

Notably	O
,	O
ARD1	B-Gene_or_gene_product
was	O
critical	O
for	O
transcriptionally	O
regulating	O
a	O
number	O
of	O
AR	B-Gene_or_gene_product
target	O
genes	O
that	O
are	O
involved	O
in	O
prostate	B-Organ
tumorigenesis	O
.	O

Furthermore	O
,	O
ARD1	B-Gene_or_gene_product
interacted	O
physically	O
with	O
and	O
acetylated	O
the	O
AR	B-Gene_or_gene_product
protein	O
in	O
vivo	O
and	O
in	O
vitro	O
.	O

Because	O
AR	B-Gene_or_gene_product
-	O
ARD1	B-Gene_or_gene_product
interaction	O
facilitated	O
the	O
AR	B-Gene_or_gene_product
binding	O
to	O
its	O
targeted	O
promoters	O
for	O
gene	O
transcription	O
,	O
we	O
propose	O
that	O
ARD1	B-Gene_or_gene_product
functions	O
as	O
a	O
unique	O
AR	B-Gene_or_gene_product
regulator	O
and	O
forms	O
a	O
positive	O
feedback	O
loop	O
for	O
AR	B-Gene_or_gene_product
-	O
dependent	O
prostate	B-Organ
tumorigenesis	O
.	O

Disruption	O
of	O
AR	B-Gene_or_gene_product
-	O
ARD1	B-Gene_or_gene_product
interactions	O
may	O
be	O
a	O
potent	O
intervention	O
for	O
androgen	B-Gene_or_gene_product
-	O
dependent	O
PCa	B-Cancer
therapy	O
.	O

The	O
role	O
of	O
Hes	B-Gene_or_gene_product
genes	O
in	O
intestinal	B-Multi-tissue_structure
development	O
,	O
homeostasis	O
and	O
tumor	B-Cancer
formation	O
.	O

Notch	B-Gene_or_gene_product
signaling	O
regulates	O
intestinal	B-Multi-tissue_structure
development	O
,	O
homeostasis	O
and	O
tumorigenesis	O
,	O
but	O
its	O
precise	O
downstream	O
mechanism	O
remains	O
largely	O
unknown	O
.	O

Here	O
we	O
found	O
that	O
inactivation	O
of	O
the	O
Notch	B-Gene_or_gene_product
effectors	O
Hes1	B-Gene_or_gene_product
,	O
Hes3	B-Gene_or_gene_product
and	O
Hes5	B-Gene_or_gene_product
,	O
but	O
not	O
Hes1	B-Gene_or_gene_product
alone	O
,	O
led	O
to	O
reduced	O
cell	B-Cell
proliferation	O
,	O
increased	O
secretory	B-Cell
cell	I-Cell
formation	O
and	O
altered	O
intestinal	B-Tissue
structures	I-Tissue
in	O
adult	B-Organism
mice	I-Organism
.	O

However	O
,	O
in	O
Apc	B-Gene_or_gene_product
mutation	O
-	O
induced	O
intestinal	B-Cancer
tumors	I-Cancer
,	O
inactivation	O
of	O
Hes1	B-Gene_or_gene_product
alone	O
was	O
sufficient	O
for	O
reducing	O
tumor	B-Cell
cell	I-Cell
proliferation	O
and	O
inducing	O
differentiation	O
of	O
tumor	B-Cell
cells	I-Cell
into	O
all	O
types	O
of	O
intestinal	B-Cell
epithelial	I-Cell
cells	I-Cell
,	O
but	O
without	O
affecting	O
the	O
homeostasis	O
of	O
normal	O
crypts	B-Multi-tissue_structure
owing	O
to	O
genetic	O
redundancy	O
.	O

These	O
results	O
indicated	O
that	O
Hes	B-Gene_or_gene_product
genes	O
cooperatively	O
regulate	O
intestinal	B-Multi-tissue_structure
development	O
and	O
homeostasis	O
and	O
raised	O
the	O
possibility	O
that	O
Hes1	B-Gene_or_gene_product
is	O
a	O
promising	O
target	O
to	O
induce	O
the	O
differentiation	O
of	O
tumor	B-Cell
cells	I-Cell
.	O

Activation	B-Gene_or_gene_product
-	I-Gene_or_gene_product
induced	I-Gene_or_gene_product
cytidine	I-Gene_or_gene_product
deaminase	I-Gene_or_gene_product
in	O
antibody	O
diversification	O
and	O
chromosome	B-Cellular_component
translocation	O
.	O

DNA	B-Cellular_component
damage	O
,	O
rearrangement	O
,	O
and	O
mutation	O
of	O
the	O
human	B-Organism
genome	O
are	O
the	O
basis	O
of	O
carcinogenesis	O
and	O
thought	O
to	O
be	O
avoided	O
at	O
all	O
costs	O
.	O

An	O
exception	O
is	O
the	O
adaptive	O
immune	B-Anatomical_system
system	I-Anatomical_system
where	O
lymphocytes	B-Cell
utilize	O
programmed	O
DNA	B-Cellular_component
damage	O
to	O
effect	O
antigen	O
receptor	O
diversification	O
.	O

Both	O
B	B-Cell
and	O
T	B-Cell
lymphocytes	I-Cell
diversify	O
their	O
antigen	O
receptors	O
through	O
RAG1	B-Gene_or_gene_product
/	O
2	B-Gene_or_gene_product
mediated	O
recombination	O
,	O
but	O
B	B-Cell
cells	I-Cell
undergo	O
two	O
additional	O
processes	O
-	O
-	O
somatic	O
hypermutation	O
(	O
SHM	O
)	O
and	O
class	O
-	O
switch	O
recombination	O
(	O
CSR	O
)	O
,	O
both	O
initiated	O
by	O
activation	B-Gene_or_gene_product
-	I-Gene_or_gene_product
induced	I-Gene_or_gene_product
cytidine	I-Gene_or_gene_product
deaminase	I-Gene_or_gene_product
(	O
AID	B-Gene_or_gene_product
)	O
.	O

AID	B-Gene_or_gene_product
deaminates	O
cytidines	B-Simple_chemical
in	O
DNA	B-Cellular_component
resulting	O
in	O
U	O
:	O
G	O
mismatches	O
that	O
are	O
processed	O
into	O
point	O
mutations	O
in	O
SHM	O
or	O
double	O
-	O
strand	O
breaks	O
in	O
CSR	O
.	O

Although	O
AID	B-Gene_or_gene_product
activity	O
is	O
focused	O
at	O
Immunoglobulin	B-Gene_or_gene_product
(	O
Ig	B-Gene_or_gene_product
)	O
gene	O
loci	O
,	O
it	O
also	O
targets	O
a	O
wide	O
array	O
of	O
non	O
-	O
Ig	B-Gene_or_gene_product
genes	O
including	O
oncogenes	O
associated	O
with	O
lymphomas	B-Cancer
.	O

Here	O
,	O
we	O
review	O
the	O
molecular	O
basis	O
of	O
AID	B-Gene_or_gene_product
regulation	O
,	O
targeting	O
,	O
and	O
initiation	O
of	O
CSR	O
and	O
SHM	O
,	O
as	O
well	O
as	O
AID	B-Gene_or_gene_product
'	O
s	O
role	O
in	O
generating	O
chromosome	B-Cellular_component
translocations	O
that	O
contribute	O
to	O
lymphomagenesis	O
.	O

Incidence	O
of	O
BRAF	B-Gene_or_gene_product
p	O
.	O
Val600Glu	B-Amino_acid
and	O
p	O
.	O
Val600Lys	B-Amino_acid
mutations	O
in	O
a	O
consecutive	O
series	O
of	O
183	O
metastatic	O
melanoma	B-Cancer
patients	B-Organism
from	O
a	O
high	O
incidence	O
region	O
.	O

AIM	O
:	O
Approximately	O
40	O
-	O
60	O
%	O
of	O
melanomas	B-Cancer
from	O
Caucasian	O
populations	O
carry	O
activating	O
mutations	O
in	O
the	O
BRAF	B-Gene_or_gene_product
oncogene	O
,	O
with	O
the	O
most	O
common	O
being	O
the	O
p	O
.	O
Val600Glu	B-Amino_acid
(	O
V600E	B-Amino_acid
)	O
hotspot	O
mutation	O
in	O
exon	O
15	O
.	O

The	O
aim	O
of	O
the	O
present	O
study	O
was	O
to	O
investigate	O
the	O
frequency	O
of	O
the	O
less	O
common	O
p	O
.	O
Val600Lys	B-Amino_acid
(	O
V600K	B-Amino_acid
)	O
mutation	O
in	O
metastatic	O
melanoma	B-Cancer
from	O
a	O
high	O
incidence	O
region	O
.	O

METHOD	O
:	O
Dideoxy	O
sequencing	O
and	O
fluorescent	O
single	O
strand	O
conformation	O
analysis	O
were	O
used	O
to	O
screen	O
for	O
mutations	O
in	O
exon	O
15	O
of	O
BRAF	B-Gene_or_gene_product
in	O
183	O
cases	O
of	O
metastatic	B-Cancer
melanoma	I-Cancer
.	O

RESULTS	O
:	O
The	O
overall	O
incidence	O
of	O
BRAF	B-Gene_or_gene_product
mutation	O
(	O
89	O
/	O
183	O
,	O
49	O
%	O
)	O
was	O
very	O
similar	O
to	O
other	O
large	O
studies	O
of	O
Caucasian	O
populations	O
.	O

However	O
,	O
the	O
frequency	O
of	O
the	O
p	O
.	O
Val600Lys	B-Amino_acid
mutation	O
was	O
higher	O
than	O
in	O
most	O
other	O
studies	O
and	O
comprised	O
almost	O
one	O
-	O
third	O
of	O
all	O
BRAF	B-Gene_or_gene_product
mutations	O
in	O
our	O
cohort	O
(	O
27	O
/	O
89	O
,	O
30	O
%	O
)	O
.	O

CONCLUSION	O
:	O
BRAF	B-Gene_or_gene_product
p	O
.	O
Val600Lys	B-Amino_acid
mutations	O
were	O
present	O
at	O
a	O
relatively	O
high	O
frequency	O
in	O
this	O
cohort	O
of	O
metastatic	O
melanoma	B-Cancer
patients	B-Organism
(	O
27	O
/	O
183	O
,	O
15	O
%	O
)	O
.	O

Assays	O
used	O
to	O
screen	O
for	O
BRAF	B-Gene_or_gene_product
mutations	O
in	O
the	O
clinic	O
should	O
be	O
robust	O
enough	O
to	O
detect	O
the	O
p	O
.	O
Val600Lys	B-Amino_acid
mutation	O
,	O
as	O
this	O
may	O
have	O
therapeutic	O
implications	O
.	O

Alterations	O
in	O
carbohydrate	B-Simple_chemical
metabolism	O
in	O
canine	B-Cancer
lymphoma	I-Cancer
.	O

Following	O
an	O
overnight	O
fast	O
,	O
blood	B-Organism_substance
samples	I-Organism_substance
were	O
obtained	O
from	O
14	O
dogs	B-Organism
with	O
previously	O
untreated	O
lymphoma	B-Cancer
before	O
and	O
5	O
,	O
15	O
,	O
30	O
,	O
45	O
,	O
60	O
,	O
and	O
90	O
minutes	O
following	O
an	O
intravenous	B-Multi-tissue_structure
challenge	O
with	O
500	O
mg	O
/	O
kg	O
dextrose	B-Simple_chemical
.	O

Samples	B-Organism_substance
were	O
assayed	O
for	O
glucose	B-Simple_chemical
,	O
lactate	B-Simple_chemical
,	O
and	O
insulin	B-Gene_or_gene_product
concentrations	O
and	O
compared	O
statistically	O
with	O
ten	O
control	O
dogs	B-Organism
of	O
similar	O
weight	O
and	O
age	O
undergoing	O
an	O
identical	O
dextrose	B-Simple_chemical
challenge	O
.	O

Dogs	B-Organism
with	O
lymphoma	B-Cancer
had	O
similar	O
glucose	B-Simple_chemical
tolerance	O
curves	O
when	O
compared	O
with	O
controls	O
.	O

Lactate	B-Simple_chemical
concentrations	O
were	O
significantly	O
higher	O
(	O
P	O
less	O
than	O
0	O
.	O
001	O
)	O
at	O
baseline	O
and	O
all	O
time	O
periods	O
of	O
the	O
glucose	B-Simple_chemical
tolerance	O
test	O
in	O
dogs	B-Organism
with	O
lymphoma	B-Cancer
when	O
compared	O
with	O
controls	O
.	O

Rise	O
in	O
lactate	B-Simple_chemical
concentrations	O
over	O
baseline	O
levels	O
in	O
the	O
first	O
30	O
minutes	O
of	O
the	O
glucose	B-Simple_chemical
tolerance	O
test	O
were	O
significantly	O
higher	O
in	O
dogs	B-Organism
with	O
lymphoma	B-Cancer
(	O
P	O
=	O
0	O
.	O
011	O
)	O
.	O

Insulin	B-Gene_or_gene_product
concentrations	O
were	O
significantly	O
higher	O
(	O
P	O
less	O
than	O
0	O
.	O
001	O
)	O
at	O
baseline	O
and	O
at	O
the	O
5	O
-	O
,	O
45	O
-	O
,	O
60	O
-	O
,	O
and	O
90	O
-	O
minute	O
time	O
periods	O
of	O
the	O
glucose	B-Simple_chemical
tolerance	O
test	O
in	O
dogs	B-Organism
with	O
lymphoma	B-Cancer
.	O

Rise	O
in	O
insulin	B-Gene_or_gene_product
concentrations	O
over	O
baseline	O
in	O
the	O
first	O
5	O
minutes	O
of	O
the	O
glucose	B-Simple_chemical
tolerance	O
test	O
were	O
also	O
significantly	O
greater	O
in	O
dogs	B-Organism
with	O
lymphoma	B-Cancer
(	O
P	O
=	O
0	O
.	O
021	O
)	O
.	O

These	O
results	O
indicate	O
carbohydrate	B-Simple_chemical
metabolism	O
is	O
altered	O
in	O
dogs	B-Organism
with	O
lymphoma	B-Cancer
.	O

Many	O
of	O
these	O
alterations	O
parallel	O
those	O
observed	O
in	O
human	B-Organism
patients	I-Organism
suffering	O
from	O
cancer	B-Organism_substance
cachexia	I-Organism_substance
making	O
canine	B-Organism
lymphoma	B-Cancer
a	O
potential	O
model	O
for	O
further	O
study	O
of	O
the	O
pathogenesis	O
and	O
therapy	O
of	O
cancer	B-Organism_substance
cachexia	I-Organism_substance
.	O

Members	O
of	O
the	O
src	B-Gene_or_gene_product
and	O
ras	B-Gene_or_gene_product
oncogene	O
families	O
supplant	O
the	O
epidermal	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
requirement	O
of	O
BALB	B-Organism
/	I-Organism
MK	I-Organism
-	I-Organism
2	I-Organism
keratinocytes	B-Cell
and	O
induce	O
distinct	O
alterations	O
in	O
their	O
terminal	O
differentiation	O
program	O
.	O

BALB	B-Organism
-	I-Organism
/	I-Organism
MK	I-Organism
-	I-Organism
2	I-Organism
mouse	I-Organism
epidermal	B-Cell
keratinocytes	I-Cell
required	O
epidermal	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
for	O
proliferation	O
and	O
terminally	O
differentiated	O
in	O
response	O
to	O
high	O
Ca2	B-Simple_chemical
+	O
concentration	O
.	O

Infection	O
with	O
retroviruses	B-Organism
containing	O
transforming	O
genes	O
of	O
the	O
src	B-Gene_or_gene_product
and	O
ras	B-Gene_or_gene_product
oncogene	O
families	O
led	O
to	O
rapid	O
loss	O
of	O
epidermal	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
dependence	O
,	O
in	O
some	O
cases	O
,	O
accompanied	O
by	O
alterations	O
in	O
cellular	B-Cell
morphology	O
.	O

The	O
virus	B-Organism
-	O
altered	O
cells	B-Cell
continued	O
to	O
proliferate	O
in	O
the	O
presence	O
of	O
high	O
levels	O
of	O
extracellular	B-Immaterial_anatomical_entity
calcium	B-Simple_chemical
but	O
exhibited	O
alterations	O
in	O
normal	O
keratinocyte	B-Cell
terminal	O
differentiation	O
that	O
appear	O
to	O
be	O
specific	O
to	O
the	O
particular	O
oncogene	O
.	O

These	O
alterations	O
bore	O
similarities	O
to	O
abnormalities	O
in	O
differentiation	O
observed	O
in	O
naturally	O
occurring	O
squamous	B-Cancer
epithelial	I-Cancer
malignancies	I-Cancer
.	O

Experimental	O
drug	O
therapy	O
of	O
peritumoral	B-Pathological_formation
brain	I-Pathological_formation
edema	I-Pathological_formation
.	O

Four	O
drugs	O
with	O
potential	O
anti	O
-	O
peritumoral	B-Pathological_formation
brain	I-Pathological_formation
edema	I-Pathological_formation
activity	O
were	O
studied	O
using	O
the	O
VX2	B-Cancer
rabbit	B-Organism
brain	B-Cancer
tumor	I-Cancer
model	O
.	O

Meclofenamate	B-Simple_chemical
and	O
indomethacin	B-Simple_chemical
were	O
tested	O
in	O
an	O
attempt	O
to	O
confirm	O
recent	O
reports	O
of	O
anti	O
-	O
edema	B-Pathological_formation
activity	O
in	O
non	B-Simple_chemical
steroidal	I-Simple_chemical
anti	I-Simple_chemical
-	I-Simple_chemical
inflammatory	I-Simple_chemical
drugs	I-Simple_chemical
(	O
NSAID	B-Simple_chemical
'	I-Simple_chemical
s	I-Simple_chemical
)	O
.	O

The	O
'	O
angiostatic	O
'	O
steroids	O
17	B-Simple_chemical
hydroxyprogesterone	I-Simple_chemical
and	O
epicortisol	B-Simple_chemical
were	O
tested	O
because	O
of	O
their	O
lack	O
of	O
glucocorticoid	B-Simple_chemical
and	O
mineralocorticoid	B-Simple_chemical
effects	O
and	O
their	O
structural	O
similarity	O
to	O
glucocorticoids	B-Simple_chemical
.	O

The	O
protein	O
and	O
water	O
component	O
of	O
brain	B-Pathological_formation
edema	I-Pathological_formation
were	O
indirectly	O
quantitated	O
.	O

None	O
of	O
the	O
test	O
drugs	O
demonstrated	O
significant	O
anti	O
-	O
edema	B-Pathological_formation
activity	O
.	O

This	O
work	O
does	O
not	O
confirm	O
reports	O
that	O
NSAID	B-Simple_chemical
'	O
s	O
have	O
anti	O
-	O
edema	B-Pathological_formation
activity	O
and	O
suggests	O
that	O
there	O
may	O
be	O
no	O
correlation	O
between	O
'	O
angiostatic	O
'	O
and	O
anti	O
-	O
edema	B-Pathological_formation
activity	O
in	O
certain	O
steroid	B-Simple_chemical
compounds	I-Simple_chemical
.	O

Identification	O
of	O
a	O
small	O
region	O
of	O
the	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
fos	I-Gene_or_gene_product
gene	O
product	O
that	O
is	O
sufficient	O
for	O
transforming	O
potential	O
and	O
growth	O
-	O
stimulating	O
activity	O
.	O

To	O
analyze	O
the	O
structure	O
-	O
function	O
relationship	O
for	O
the	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
fos	I-Gene_or_gene_product
protein	O
,	O
we	O
constructed	O
in	O
-	O
frame	O
insertion	O
and	O
deletion	O
mutants	O
of	O
the	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
fos	I-Gene_or_gene_product
gene	O
carried	O
by	O
FBJ	B-Organism
-	I-Organism
MuSV	I-Organism
,	O
and	O
expressed	O
them	O
in	O
chicken	B-Organism
primary	O
cells	B-Cell
using	O
retrovirus	O
vectors	O
.	O

We	O
assessed	O
the	O
effects	O
of	O
these	O
mutations	O
on	O
the	O
ability	O
of	O
the	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
fos	I-Gene_or_gene_product
protein	O
to	O
transform	O
chicken	B-Organism
embryo	B-Cell
fibroblasts	I-Cell
and	O
to	O
stimulate	O
cellular	B-Cell
proliferation	O
of	O
chicken	B-Organism
neuroretinal	B-Cell
cells	I-Cell
.	O

The	O
mutant	O
which	O
retains	O
only	O
the	O
central	O
region	O
of	O
the	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
fos	I-Gene_or_gene_product
protein	O
(	O
Met111	B-Amino_acid
-	O
Ile206	B-Amino_acid
)	O
have	O
both	O
activities	O
,	O
but	O
the	O
mutants	O
which	O
have	O
deletions	O
in	O
this	O
region	O
,	O
and	O
one	O
of	O
the	O
mutants	O
that	O
has	O
a	O
four	O
amino	B-Amino_acid
acid	I-Amino_acid
insertion	O
in	O
it	O
,	O
lost	O
both	O
activities	O
.	O

The	O
central	O
region	O
that	O
is	O
sufficient	O
for	O
these	O
activities	O
includes	O
the	O
evolutionarily	O
highly	O
conserved	O
region	O
among	O
human	B-Organism
,	O
mouse	B-Organism
and	O
chicken	B-Organism
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
fos	I-Gene_or_gene_product
proteins	O
.	O

Additionally	O
,	O
this	O
sequence	O
shares	O
some	O
homology	O
with	O
the	O
DNA	B-Cellular_component
binding	O
domain	O
of	O
GCN4	B-Gene_or_gene_product
and	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
jun	I-Gene_or_gene_product
protein	O
.	O

The	O
truncated	O
fos	B-Gene_or_gene_product
protein	O
that	O
contains	O
only	O
part	O
of	O
the	O
central	O
region	O
is	O
not	O
phosphorylated	O
in	O
chicken	B-Organism
embryo	B-Cell
fibroblasts	I-Cell
,	O
indicating	O
that	O
phosphorylation	O
of	O
the	O
fos	B-Gene_or_gene_product
protein	O
is	O
not	O
necessary	O
for	O
the	O
transforming	O
activity	O
.	O

Inhibition	O
by	O
retinoic	B-Simple_chemical
acid	I-Simple_chemical
of	O
type	O
IV	O
collagenolysis	O
and	O
invasion	O
through	O
reconstituted	O
basement	B-Cellular_component
membrane	I-Cellular_component
by	O
metastatic	O
rat	B-Organism
mammary	B-Cell
adenocarcinoma	I-Cell
cells	I-Cell
.	O

The	O
activity	O
of	O
type	B-Gene_or_gene_product
IV	I-Gene_or_gene_product
collagenase	I-Gene_or_gene_product
,	O
which	O
enables	O
tumor	B-Cell
cells	I-Cell
to	O
degrade	O
collagen	B-Gene_or_gene_product
type	I-Gene_or_gene_product
IV	I-Gene_or_gene_product
found	O
in	O
the	O
subendothelial	B-Cellular_component
basement	I-Cellular_component
membrane	I-Cellular_component
,	O
has	O
been	O
correlated	O
with	O
the	O
metastatic	O
potential	O
in	O
several	O
tumor	B-Cancer
types	O
,	O
including	O
the	O
rat	B-Organism
13762NF	B-Cell
mammary	I-Cell
adenocarcinoma	I-Cell
cell	I-Cell
line	I-Cell
and	O
its	O
clones	B-Cell
.	O

In	O
this	O
study	O
,	O
we	O
examined	O
whether	O
all	B-Simple_chemical
-	I-Simple_chemical
trans	I-Simple_chemical
-	I-Simple_chemical
retinoic	I-Simple_chemical
acid	I-Simple_chemical
(	O
all	B-Simple_chemical
-	I-Simple_chemical
trans	I-Simple_chemical
-	I-Simple_chemical
RA	I-Simple_chemical
)	O
and	O
other	O
retinoids	O
,	O
which	O
exhibit	O
antitumor	B-Cancer
activity	O
in	O
vitro	O
and	O
in	O
vivo	O
,	O
affect	O
the	O
collagenolytic	O
activity	O
of	O
metastatic	O
rat	B-Organism
13762NF	B-Cell
mammary	I-Cell
adenocarcinoma	I-Cell
cells	I-Cell
.	O

Cells	B-Cell
of	O
the	O
highly	O
metastatic	O
lung	B-Organ
-	O
colonizing	O
clone	B-Cell
MTF7	I-Cell
.	I-Cell
T35	I-Cell
.	I-Cell
3	I-Cell
,	O
derived	O
from	O
the	O
13762NF	B-Cell
cell	I-Cell
line	I-Cell
,	O
were	O
treated	O
for	O
3	O
days	O
with	O
0	O
.	O
1	O
,	O
1	O
,	O
or	O
10	O
microM	O
all	B-Simple_chemical
-	I-Simple_chemical
trans	I-Simple_chemical
-	I-Simple_chemical
RA	I-Simple_chemical
,	O
harvested	O
,	O
and	O
seeded	O
on	O
[	B-Simple_chemical
3H	I-Simple_chemical
]	I-Simple_chemical
proline	I-Simple_chemical
-	O
labeled	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
deposited	O
by	O
cultured	O
rat	B-Organism
lung	B-Cell
endothelial	I-Cell
cells	I-Cell
or	O
on	O
a	O
film	O
of	O
purified	O
[	B-Simple_chemical
3H	I-Simple_chemical
]	I-Simple_chemical
proline	I-Simple_chemical
-	O
labeled	O
type	B-Gene_or_gene_product
IV	I-Gene_or_gene_product
collagen	I-Gene_or_gene_product
.	O

The	O
amount	O
of	O
radioactivity	O
released	O
into	O
the	O
medium	O
during	O
the	O
subsequent	O
24	O
to	O
72	O
h	O
was	O
measured	O
,	O
and	O
it	O
was	O
found	O
that	O
all	B-Simple_chemical
-	I-Simple_chemical
trans	I-Simple_chemical
-	I-Simple_chemical
RA	I-Simple_chemical
treatment	O
inhibited	O
degradation	O
of	O
extracellular	B-Cellular_component
matrix	I-Cellular_component
and	O
type	B-Gene_or_gene_product
IV	I-Gene_or_gene_product
collagen	I-Gene_or_gene_product
by	O
50	O
to	O
60	O
%	O
.	O

This	O
effect	O
was	O
observed	O
whether	O
the	O
cells	B-Cell
had	O
been	O
treated	O
with	O
all	B-Simple_chemical
-	I-Simple_chemical
trans	I-Simple_chemical
-	I-Simple_chemical
RA	I-Simple_chemical
in	O
serum	B-Organism_substance
-	O
free	O
medium	O
or	O
in	O
medium	O
supplemented	O
with	O
heat	O
-	O
inactivated	O
or	O
acid	O
-	O
treated	O
fetal	B-Developing_anatomical_structure
bovine	B-Organism
serum	B-Organism_substance
.	O

The	O
growth	O
of	O
the	O
cells	B-Cell
was	O
not	O
inhibited	O
under	O
these	O
conditions	O
,	O
except	O
after	O
treatment	O
with	O
10	O
microM	O
all	B-Simple_chemical
-	I-Simple_chemical
trans	I-Simple_chemical
-	I-Simple_chemical
RA	I-Simple_chemical
in	O
serum	B-Organism_substance
-	O
free	O
medium	O
.	O

The	O
reduction	O
in	O
collagenolytic	O
activity	O
was	O
observed	O
in	O
viable	O
cells	B-Cell
as	O
well	O
as	O
in	O
conditioned	O
medium	O
.	O

A	O
24	O
-	O
h	O
exposure	O
of	O
cells	B-Cell
to	O
all	B-Simple_chemical
-	I-Simple_chemical
trans	I-Simple_chemical
-	I-Simple_chemical
RA	I-Simple_chemical
was	O
sufficient	O
to	O
cause	O
a	O
30	O
%	O
decrease	O
in	O
the	O
collagenolytic	O
activity	O
,	O
and	O
this	O
inhibitory	O
effect	O
was	O
reversible	O
.	O

The	O
direct	O
addition	O
of	O
all	B-Simple_chemical
-	I-Simple_chemical
trans	I-Simple_chemical
-	I-Simple_chemical
RA	I-Simple_chemical
to	O
conditioned	O
medium	O
had	O
no	O
effect	O
on	O
secreted	O
collagenase	B-Gene_or_gene_product
activity	O
.	O

The	O
apparent	O
molecular	O
weights	O
of	O
the	O
collagenolytic	O
enzymes	O
were	O
determined	O
by	O
electrophoresis	O
of	O
cell	B-Organism_substance
extracts	I-Organism_substance
and	O
concentrated	O
conditioned	O
medium	O
in	O
type	B-Gene_or_gene_product
IV	I-Gene_or_gene_product
collagen	I-Gene_or_gene_product
-	O
embedded	O
polyacrylamide	B-Simple_chemical
gels	O
followed	O
by	O
renaturation	O
and	O
activation	O
of	O
the	O
enzymes	O
within	O
the	O
gels	O
.	O

Two	O
major	O
type	O
IV	O
collagenolytic	O
metalloproteinases	B-Gene_or_gene_product
exhibiting	O
molecular	O
weights	O
of	O
64	O
,	O
000	O
and	O
88	O
,	O
000	O
,	O
respectively	O
,	O
were	O
detected	O
by	O
this	O
method	O
.	O

These	O
two	O
enzymes	O
were	O
also	O
found	O
to	O
have	O
specificity	O
for	O
gelatin	O
.	O

The	O
Mr	O
64	O
,	O
000	O
enzyme	O
could	O
be	O
extracted	O
from	O
viable	O
cells	B-Cell
(	O
presumably	O
from	O
the	O
cell	B-Cellular_component
membrane	I-Cellular_component
)	O
by	O
2	O
%	O
1	B-Simple_chemical
-	I-Simple_chemical
butanol	I-Simple_chemical
.	O

Treatment	O
with	O
all	B-Simple_chemical
-	I-Simple_chemical
trans	I-Simple_chemical
-	I-Simple_chemical
RA	I-Simple_chemical
decreased	O
the	O
level	O
of	O
these	O
enzymes	O
in	O
the	O
cellular	B-Cell
,	O
cell	B-Cellular_component
membrane	I-Cellular_component
,	O
and	O
conditioned	O
medium	O
compartments	O
.	O
(	O
ABSTRACT	O
TRUNCATED	O
AT	O
400	O
WORDS	O
)	O

Malate	B-Simple_chemical
-	O
citrate	B-Simple_chemical
cycle	O
during	O
glycolysis	O
and	O
glutaminolysis	O
in	O
Ehrlich	B-Cell
ascites	I-Cell
tumor	I-Cell
cells	I-Cell
.	O

The	O
malate	B-Simple_chemical
-	O
citrate	B-Simple_chemical
cycle	O
was	O
studied	O
during	O
aerobic	O
glycolysis	O
and	O
glutaminolysis	O
in	O
a	O
strain	O
of	O
Ehrlich	B-Cell
ascites	I-Cell
tumor	I-Cell
cells	I-Cell
which	O
showed	O
a	O
very	O
low	O
malate	B-Simple_chemical
-	O
aspartate	B-Simple_chemical
shuttle	O
system	O
activity	O
.	O

The	O
experimental	O
approach	O
includes	O
:	O
estimation	O
of	O
mitochondrial	B-Cellular_component
NAD	B-Simple_chemical
[	I-Simple_chemical
P	I-Simple_chemical
]	I-Simple_chemical
+	O
-	O
dependent	O
malic	O
enzyme	O
activity	O
;	O
respiratory	O
activity	O
of	O
freshly	O
harvested	O
or	O
fasted	O
cells	B-Cell
,	O
and	O
of	O
isolated	O
mitochondria	B-Cellular_component
;	O
and	O
determination	O
of	O
the	O
metabolites	O
involved	O
in	O
the	O
glycolytic	O
and	O
glutaminolytic	O
pathways	O
.	O

The	O
results	O
suggest	O
that	O
in	O
this	O
strain	B-Cell
,	O
the	O
malate	B-Simple_chemical
-	O
citrate	B-Simple_chemical
shuttle	O
is	O
not	O
an	O
effective	O
pathway	O
for	O
transferring	O
glycolytic	O
reducing	O
equivalents	O
from	O
cytosol	B-Organism_substance
to	O
mitochondria	B-Cellular_component
.	O

Less	O
than	O
15	O
%	O
of	O
the	O
glucose	B-Simple_chemical
uptake	O
was	O
affected	O
by	O
the	O
1	B-Simple_chemical
,	I-Simple_chemical
2	I-Simple_chemical
,	I-Simple_chemical
3	I-Simple_chemical
-	I-Simple_chemical
benzenetricarboxylate	I-Simple_chemical
inhibition	O
of	O
the	O
malate	B-Simple_chemical
-	O
citrate	B-Simple_chemical
shuttle	O
.	O

Moreover	O
,	O
in	O
the	O
presence	O
of	O
glucose	B-Simple_chemical
,	O
the	O
malate	B-Simple_chemical
-	O
citrate	B-Simple_chemical
cycle	O
did	O
not	O
appear	O
to	O
play	O
an	O
important	O
role	O
in	O
the	O
glutaminolytic	O
process	O
.	O

The	O
present	O
work	O
supports	O
and	O
extends	O
the	O
finding	O
of	O
previous	O
studies	O
,	O
since	O
the	O
results	O
showed	O
that	O
the	O
glucose	B-Simple_chemical
metabolism	O
depressed	O
the	O
oxidative	O
processes	O
in	O
Ehrlich	B-Cancer
ascites	I-Cancer
tumor	I-Cancer
mitochondria	B-Cellular_component
,	O
not	O
only	O
alone	O
,	O
but	O
also	O
in	O
the	O
presence	O
of	O
glutamine	B-Amino_acid
.	O

Interestingly	O
,	O
the	O
high	O
glutamine	B-Amino_acid
uptake	O
was	O
maintained	O
in	O
the	O
presence	O
of	O
glucose	B-Simple_chemical
.	O

N	B-Simple_chemical
-	I-Simple_chemical
methyl	I-Simple_chemical
-	I-Simple_chemical
N	I-Simple_chemical
-	I-Simple_chemical
nitrosourea	I-Simple_chemical
-	O
induced	O
transformation	O
of	O
rat	B-Organism
urothelial	B-Cell
cells	I-Cell
in	O
vitro	O
is	O
not	O
mediated	O
by	O
activation	O
of	O
ras	B-Gene_or_gene_product
oncogenes	O
.	O

Adult	O
rat	B-Organism
urothelial	B-Cell
cells	I-Cell
were	O
transformed	O
in	O
vitro	O
following	O
treatment	O
with	O
a	O
single	O
dose	O
of	O
N	B-Simple_chemical
-	I-Simple_chemical
methyl	I-Simple_chemical
-	I-Simple_chemical
N	I-Simple_chemical
-	I-Simple_chemical
nitrosourea	I-Simple_chemical
(	O
MNU	B-Simple_chemical
)	O
or	O
MNU	B-Simple_chemical
treatment	O
followed	O
by	O
promotion	O
with	O
sodium	B-Simple_chemical
saccharin	I-Simple_chemical
.	O

This	O
in	O
vitro	O
transformation	O
process	O
involves	O
multiple	O
steps	O
:	O
slow	O
-	O
growing	O
'	O
pre	O
-	O
neoplastic	O
'	O
epithelial	B-Cell
foci	I-Cell
are	O
induced	O
70	O
-	O
100	O
days	O
after	O
MNU	B-Simple_chemical
treatment	O
and	O
from	O
such	O
foci	B-Cell
rapidly	O
proliferating	O
immortal	O
cell	B-Cell
lines	I-Cell
were	O
established	O
,	O
some	O
of	O
which	O
became	O
tumorigenic	O
after	O
a	O
further	O
latent	O
period	O
.	O

A	O
series	O
of	O
epithelial	B-Cell
cell	I-Cell
lines	I-Cell
and	O
a	O
single	O
fibroblast	B-Cell
cell	I-Cell
line	I-Cell
established	O
in	O
this	O
way	O
were	O
analysed	O
for	O
the	O
presence	O
of	O
transforming	O
genes	O
by	O
DNA	B-Cellular_component
transfection	O
into	O
NIH3T3	B-Cell
cells	I-Cell
.	O

None	O
of	O
the	O
epithelial	B-Cell
cell	I-Cell
lines	I-Cell
induced	O
foci	B-Cell
in	O
a	O
focus	B-Cell
formation	O
assay	O
.	O

The	O
single	O
non	B-Cell
-	I-Cell
epithelial	I-Cell
line	I-Cell
induced	O
foci	B-Cell
and	O
was	O
found	O
to	O
contain	O
an	O
activated	O
c	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
gene	O
with	O
a	O
G	B-Simple_chemical
-	O
-	O
-	O
-	O
A	B-Simple_chemical
transition	O
in	O
codon	O
12	O
.	O

To	O
assay	O
for	O
the	O
possible	O
presence	O
of	O
transforming	O
genes	O
which	O
were	O
not	O
active	O
in	O
a	O
focus	B-Cell
formation	O
assay	O
,	O
two	O
of	O
the	O
epithelial	B-Cell
lines	I-Cell
were	O
analysed	O
further	O
by	O
co	O
-	O
transfection	O
with	O
a	O
dominant	O
selectable	O
marker	O
,	O
followed	O
by	O
selection	O
and	O
inoculation	O
into	O
nude	B-Organism
mice	I-Organism
.	O

No	O
tumours	B-Cancer
were	O
induced	O
within	O
the	O
latent	O
period	O
for	O
tumour	B-Cancer
production	O
by	O
control	B-Cell
cells	I-Cell
transfected	O
with	O
NIH3T3	B-Cell
cell	I-Cell
DNA	B-Cellular_component
(	O
40	O
-	O
60	O
days	O
)	O
.	O

These	O
results	O
suggest	O
that	O
there	O
is	O
cell	B-Cell
type	O
specificity	O
for	O
oncogene	O
activation	O
during	O
in	O
vitro	O
rat	B-Organism
bladder	B-Organ
transformation	O
initiated	O
by	O
a	O
single	O
carcinogen	O
and	O
that	O
ras	B-Gene_or_gene_product
gene	O
activation	O
is	O
not	O
a	O
necessary	O
step	O
in	O
urothelial	B-Cell
transformation	O
in	O
vitro	O
.	O

Low	O
-	O
level	O
X	O
-	O
radiation	O
effects	O
on	O
functional	O
vascular	B-Multi-tissue_structure
changes	O
in	O
Syrian	B-Organism
hamster	I-Organism
cheek	B-Tissue
pouch	I-Tissue
epithelium	I-Tissue
during	O
hydrocarbon	O
carcinogenesis	O
.	O

Effects	O
of	O
repeated	O
low	O
-	O
level	O
X	O
radiation	O
on	O
functional	O
microvascular	B-Tissue
changes	O
in	O
hamster	B-Organism
cheek	B-Tissue
pouch	I-Tissue
epithelium	I-Tissue
during	O
and	O
following	O
carcinogenesis	O
by	O
7	B-Simple_chemical
,	I-Simple_chemical
12	I-Simple_chemical
-	I-Simple_chemical
dimethylbenz	I-Simple_chemical
[	I-Simple_chemical
a	I-Simple_chemical
]	I-Simple_chemical
anthracene	I-Simple_chemical
(	O
DMBA	B-Simple_chemical
)	O
were	O
studied	O
.	O

Prior	O
studies	O
showed	O
enhancement	O
of	O
such	O
carcinogenesis	O
by	O
repeated	O
20	O
rad	O
head	B-Organism_subdivision
and	O
neck	B-Organism_subdivision
X	O
-	O
radiation	O
exposures	O
,	O
and	O
it	O
was	O
proposed	O
that	O
one	O
possible	O
mechanism	O
was	O
radiogenic	O
alteration	O
of	O
the	O
functional	O
microvasculature	B-Tissue
in	O
a	O
manner	O
which	O
favored	O
subsequent	O
tumor	B-Cancer
development	O
.	O

Hamsters	B-Organism
were	O
treated	O
with	O
either	O
radiation	O
,	O
DMBA	B-Simple_chemical
,	O
radiation	O
+	O
DMBA	B-Simple_chemical
,	O
or	O
no	O
treatment	O
.	O

Animals	B-Organism
were	O
sacrificed	O
at	O
3	O
-	O
week	O
intervals	O
from	O
0	O
to	O
39	O
weeks	O
after	O
treatments	O
began	O
.	O

Pouch	B-Multi-tissue_structure
vascular	I-Multi-tissue_structure
volume	O
and	O
permeability	O
changes	O
were	O
studied	O
by	O
fractional	O
distributions	O
of	O
radiotracers	O
and	O
were	O
analyzed	O
by	O
a	O
variety	O
of	O
statistical	O
methods	O
which	O
explored	O
the	O
vascular	B-Multi-tissue_structure
parameters	O
,	O
treatment	O
types	O
,	O
elapsed	O
time	O
,	O
presence	O
of	O
the	O
carcinogen	O
,	O
and	O
histopathologic	O
changes	O
.	O

All	O
treatments	O
resulted	O
in	O
significant	O
changes	O
in	O
vascular	B-Multi-tissue_structure
volume	O
with	O
time	O
,	O
while	O
only	O
DMBA	B-Simple_chemical
treatments	O
alone	O
resulted	O
in	O
significant	O
changes	O
in	O
vascular	B-Multi-tissue_structure
permeability	O
with	O
time	O
.	O

Prior	O
to	O
the	O
appearances	O
of	O
frank	O
neoplasms	B-Cancer
,	O
volumetric	O
changes	O
in	O
DMBA	B-Simple_chemical
only	O
and	O
radiation	O
only	O
groups	O
were	O
similar	O
,	O
while	O
volume	O
changes	O
in	O
DMBA	B-Simple_chemical
+	O
radiation	O
groups	O
increased	O
slowly	O
to	O
a	O
peak	O
later	O
than	O
in	O
other	O
groups	O
and	O
then	O
declined	O
steadily	O
to	O
levels	O
similar	O
to	O
the	O
radiation	O
only	O
group	O
.	O

As	O
in	O
prior	O
studies	O
,	O
there	O
were	O
significant	O
vascular	B-Multi-tissue_structure
volume	O
differences	O
between	O
DMBA	B-Simple_chemical
and	O
DMBA	B-Simple_chemical
+	O
radiation	O
groups	O
of	O
tumor	B-Cancer
-	O
bearing	O
cheek	B-Organism_subdivision
pouches	I-Organism_subdivision
.	O

DMBA	B-Simple_chemical
maxima	O
were	O
significantly	O
higher	O
than	O
those	O
of	O
DMBA	B-Simple_chemical
+	O
radiation	O
.	O

Radiation	O
significantly	O
affected	O
DMBA	B-Simple_chemical
-	O
associated	O
vascular	B-Multi-tissue_structure
volume	O
and	O
permeability	O
changes	O
during	O
carcinogenesis	O
.	O

Several	O
possible	O
explanations	O
for	O
the	O
relationship	O
of	O
these	O
changes	O
to	O
the	O
enhancement	O
of	O
DMBA	B-Simple_chemical
carcinogenesis	O
include	O
:	O
radiation	O
blocking	O
normal	O
capillary	B-Tissue
proliferative	O
and	O
/	O
or	O
dilatory	O
responses	O
to	O
inflammation	O
secondary	O
to	O
neoplastic	O
changes	O
;	O
radiation	O
-	O
induced	O
focal	O
increases	O
in	O
the	O
pericapillary	B-Tissue
connective	I-Tissue
tissue	I-Tissue
histohematic	B-Multi-tissue_structure
barrier	I-Multi-tissue_structure
,	O
stimulating	O
angiogenesis	O
but	O
reducing	O
nutrient	O
diffusion	O
;	O
radiation	O
exposures	O
sensitizing	O
vascular	B-Tissue
endothelium	I-Tissue
to	O
subsequent	O
angiogenic	O
stimulation	O
from	O
premalignant	B-Tissue
tissues	I-Tissue
;	O
DMBA	B-Simple_chemical
vascular	B-Multi-tissue_structure
and	O
epithelial	B-Tissue
effects	O
partially	O
or	O
completely	O
blocking	O
radiation	O
effects	O
on	O
epithelial	B-Cell
and	O
/	O
or	O
endothelial	B-Cell
cells	I-Cell
;	O
and	O
radiation	O
damage	O
to	O
vessel	B-Multi-tissue_structure
walls	I-Multi-tissue_structure
partially	O
or	O
fully	O
inhibiting	O
normal	O
physiologic	O
mechanisms	O
of	O
repairing	O
DMBA	B-Simple_chemical
damage	O
to	O
the	O
vessels	B-Multi-tissue_structure
.	O

Scanning	O
electron	O
microscopy	O
of	O
vascular	B-Multi-tissue_structure
casts	O
in	O
experimental	O
ocular	B-Organ
vasoproliferation	O
.	O

Scanning	O
electron	O
microscopy	O
of	O
vascular	B-Multi-tissue_structure
casts	O
was	O
used	O
to	O
investigate	O
three	O
experimental	O
models	O
of	O
neovascularization	O
.	O

In	O
each	O
experimental	O
situation	O
,	O
the	O
casts	O
provided	O
a	O
valuable	O
three	O
dimensional	O
representation	O
of	O
the	O
newly	O
formed	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
and	O
permitted	O
subclassification	O
of	O
the	O
vessels	B-Multi-tissue_structure
within	O
normal	O
and	O
proliferating	O
vascular	B-Multi-tissue_structure
networks	I-Multi-tissue_structure
.	O

They	O
defined	O
also	O
the	O
predominant	O
origin	O
of	O
new	O
vessels	B-Multi-tissue_structure
from	O
venules	B-Multi-tissue_structure
and	O
capillaries	B-Tissue
,	O
and	O
enabled	O
the	O
evolution	O
of	O
proliferating	O
vessels	B-Multi-tissue_structure
into	O
arterioles	B-Multi-tissue_structure
and	O
venules	B-Multi-tissue_structure
to	O
be	O
documented	O
.	O

Although	O
vascular	B-Multi-tissue_structure
casts	O
must	O
be	O
interpreted	O
with	O
caution	O
in	O
light	O
of	O
the	O
possibility	O
of	O
incomplete	O
filling	O
and	O
other	O
artifacts	O
,	O
they	O
are	O
a	O
valuable	O
tool	O
in	O
the	O
study	O
of	O
ocular	B-Organ
vasoproliferation	O
.	O

Participation	O
of	O
p53	B-Gene_or_gene_product
cellular	B-Cell
tumour	B-Cancer
antigen	O
in	O
transformation	O
of	O
normal	O
embryonic	B-Cell
cells	I-Cell
.	O

The	O
cellular	B-Cell
tumour	B-Cancer
antigen	O
p53	B-Gene_or_gene_product
is	O
found	O
at	O
elevated	O
levels	O
in	O
a	O
wide	O
variety	O
of	O
transformed	O
cells	B-Cell
(	O
for	O
reviews	O
see	O
refs	O
1	O
,	O
2	O
)	O
.	O

Very	O
little	O
is	O
yet	O
known	O
about	O
the	O
precise	O
relationship	O
of	O
p53	B-Gene_or_gene_product
to	O
malignant	O
transformation	O
.	O

Although	O
the	O
increase	O
in	O
p53	B-Gene_or_gene_product
levels	O
could	O
be	O
a	O
secondary	O
by	O
-	O
product	O
of	O
the	O
transformed	O
state	O
,	O
it	O
is	O
equally	O
possible	O
that	O
p53	B-Gene_or_gene_product
is	O
actively	O
involved	O
in	O
altering	O
cellular	B-Cell
growth	O
properties	O
,	O
especially	O
as	O
it	O
has	O
been	O
implicated	O
in	O
the	O
regulation	O
of	O
normal	O
cell	B-Cell
proliferation	O
.	O

We	O
sought	O
to	O
test	O
whether	O
p53	B-Gene_or_gene_product
could	O
behave	O
in	O
a	O
manner	O
similar	O
to	O
known	O
genes	O
in	O
a	O
biological	O
test	O
system	O
,	O
and	O
we	O
demonstrate	O
here	O
that	O
p53	B-Gene_or_gene_product
can	O
cooperate	O
with	O
the	O
activated	O
Ha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
oncogene	O
to	O
transform	O
normal	O
embryonic	B-Cell
cells	I-Cell
.	O

The	O
resultant	O
foci	B-Cell
contain	O
cells	B-Cell
of	O
a	O
markedly	O
altered	O
morphology	O
which	O
produce	O
high	O
levels	O
of	O
p53	B-Gene_or_gene_product
.	O

Cell	B-Cell
lines	I-Cell
established	O
from	O
such	O
foci	B-Cell
elicit	O
tumours	B-Cancer
in	O
syngeneic	O
animals	O
.	O

Interaction	O
between	O
cellular	B-Cell
and	O
viral	B-Organism
genes	O
in	O
the	O
expression	O
of	O
the	O
RSV	B-Organism
-	O
induced	O
transformation	O
-	O
specific	O
cell	B-Cellular_component
-	I-Cellular_component
surface	I-Cellular_component
antigen	O
VCSA	B-Gene_or_gene_product
.	O

Transformation	O
of	O
BHK	B-Cell
hamster	I-Cell
fibroblasts	I-Cell
by	O
an	O
env	O
-	O
strain	O
of	O
Rous	B-Organism
sarcoma	I-Organism
virus	I-Organism
(	O
RSV	B-Organism
)	O
leads	O
to	O
the	O
appearance	O
at	O
the	O
cell	B-Cellular_component
surface	I-Cellular_component
of	O
a	O
virus	B-Gene_or_gene_product
-	I-Gene_or_gene_product
induced	I-Gene_or_gene_product
nonvirion	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
(	O
VCSA	B-Gene_or_gene_product
)	O
,	O
specific	O
for	O
transformation	O
,	O
whose	O
expression	O
is	O
controlled	O
by	O
the	O
transforming	O
src	B-Gene_or_gene_product
gene	O
.	O

Previous	O
work	O
has	O
shown	O
that	O
a	O
rabbit	B-Organism
anti	O
-	O
VCSA	B-Gene_or_gene_product
serum	B-Organism_substance
lyses	I-Organism_substance
specifically	O
,	O
in	O
the	O
presence	O
of	O
complement	O
,	O
51Cr	B-Simple_chemical
-	O
labelled	O
RSV	B-Organism
-	O
transformed	O
cells	B-Cell
from	O
different	O
animal	O
species	O
.	O

Now	O
,	O
by	O
competition	O
experiments	O
with	O
a	O
panel	O
of	O
different	O
unlabelled	O
cells	B-Cell
we	O
show	O
that	O
the	O
VCSA	B-Gene_or_gene_product
expressed	O
on	O
RSV	B-Organism
-	O
transformed	O
hamster	B-Organism
fibroblasts	B-Cell
is	O
a	O
complex	O
of	O
at	O
least	O
three	O
distinct	O
antigenic	O
specificities	O
:	O
(	O
1	O
)	O
one	O
expressed	O
on	O
all	O
RSV	B-Organism
-	O
transformed	O
fibroblasts	B-Cell
,	O
regardless	O
their	O
species	O
and	O
the	O
subgroup	O
or	O
strain	O
of	O
the	O
transforming	O
virus	B-Organism
;	O
(	O
2	O
)	O
one	O
cross	O
-	O
reacting	O
with	O
a	O
cell	B-Gene_or_gene_product
-	I-Gene_or_gene_product
surface	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
(	O
CSA	B-Gene_or_gene_product
)	O
expressed	O
at	O
various	O
degrees	O
on	O
untransformed	O
avian	B-Organism
fibroblasts	B-Cell
,	O
but	O
not	O
on	O
mammalian	B-Cell
fibroblasts	I-Cell
;	O
(	O
3	O
)	O
one	O
species	O
-	O
specific	O
,	O
present	O
only	O
on	O
RSV	B-Organism
-	O
transformed	O
hamster	B-Organism
fibroblasts	B-Cell
.	O

It	O
is	O
concluded	O
that	O
VCSA	B-Gene_or_gene_product
is	O
a	O
complex	O
of	O
several	O
antigenic	O
determinants	O
,	O
and	O
that	O
some	O
of	O
these	O
differ	O
in	O
different	O
cells	B-Cell
transformed	O
by	O
RSV	B-Organism
.	O

This	O
observation	O
indicates	O
that	O
VCSA	B-Gene_or_gene_product
expression	O
at	O
the	O
cell	B-Cellular_component
surface	I-Cellular_component
is	O
likely	O
to	O
be	O
the	O
result	O
of	O
the	O
interaction	O
between	O
the	O
viral	B-Organism
src	B-Gene_or_gene_product
gene	O
product	O
pp60src	B-Gene_or_gene_product
with	O
host	O
cell	B-Cell
gene	O
(	O
s	O
)	O
or	O
gene	O
product	O
(	O
s	O
)	O
,	O
rather	O
than	O
the	O
simple	O
expression	O
of	O
this	O
molecule	O
at	O
the	O
cell	B-Cellular_component
surface	I-Cellular_component
.	O

The	O
effect	O
of	O
translocations	O
on	O
the	O
cellular	B-Cell
myc	B-Gene_or_gene_product
gene	O
in	O
Burkitt	B-Cancer
lymphomas	I-Cancer
.	O

Chromosomal	B-Cellular_component
translocations	O
are	O
found	O
to	O
be	O
a	O
characteristic	O
feature	O
of	O
Burkitt	B-Cancer
lymphomas	I-Cancer
.	O

Similar	O
translocations	O
are	O
found	O
in	O
mouse	B-Organism
plasmacytomas	B-Cancer
and	O
both	O
diseases	O
involve	O
interchanges	O
between	O
one	O
of	O
the	O
immunoglobulin	B-Gene_or_gene_product
loci	O
and	O
DNA	B-Cellular_component
in	O
the	O
vicinity	O
of	O
the	O
myc	B-Gene_or_gene_product
gene	O
.	O

The	O
structure	O
of	O
the	O
myc	B-Gene_or_gene_product
gene	O
has	O
been	O
elucidated	O
from	O
studies	O
on	O
translocated	O
versions	O
of	O
the	O
gene	O
.	O

Activation	O
of	O
the	O
myc	B-Gene_or_gene_product
gene	O
may	O
play	O
a	O
role	O
in	O
transformation	O
by	O
promoting	O
growth	O
of	O
the	O
cells	B-Cell
bearing	O
the	O
rearranged	O
chromosomes	B-Cellular_component
.	O

Inhibition	O
by	O
N	B-Simple_chemical
-	I-Simple_chemical
(	I-Simple_chemical
phosphonacetyl	I-Simple_chemical
)	I-Simple_chemical
-	I-Simple_chemical
L	I-Simple_chemical
-	I-Simple_chemical
aspartate	I-Simple_chemical
of	O
Ehrlich	B-Cancer
ascites	I-Cancer
tumour	I-Cancer
growth	O
and	O
glucose	B-Simple_chemical
transport	O
.	O

N	B-Simple_chemical
-	I-Simple_chemical
(	I-Simple_chemical
Phosphonacetyl	I-Simple_chemical
)	I-Simple_chemical
-	I-Simple_chemical
L	I-Simple_chemical
-	I-Simple_chemical
aspartate	I-Simple_chemical
(	O
PALA	B-Simple_chemical
)	O
suppressed	O
the	O
growth	O
of	O
Ehrlich	B-Cell
ascites	I-Cell
tumour	I-Cell
cells	I-Cell
in	O
vivo	O
in	O
a	O
dose	O
-	O
dependent	O
manner	O
.	O

Simultaneously	O
as	O
the	O
growth	O
rate	O
decreased	O
,	O
the	O
cellular	B-Cell
uptake	O
of	O
glucose	B-Simple_chemical
and	O
the	O
density	O
of	O
a	O
class	O
of	O
glucose	B-Simple_chemical
-	O
reversible	O
binding	O
sites	O
for	O
cytochalasin	B-Simple_chemical
B	I-Simple_chemical
on	O
the	O
cell	B-Cellular_component
surface	I-Cellular_component
were	O
also	O
found	O
to	O
be	O
reduced	O
.	O

There	O
is	O
a	O
highly	O
significant	O
correlation	O
between	O
the	O
magnitude	O
of	O
changes	O
in	O
the	O
number	O
of	O
cytochalasin	B-Simple_chemical
B	I-Simple_chemical
binding	O
sites	O
and	O
the	O
magnitude	O
of	O
changes	O
in	O
glucose	B-Simple_chemical
uptake	O
.	O

The	O
physiological	O
significance	O
of	O
these	O
observations	O
are	O
discussed	O
.	O

Effects	O
of	O
3	B-Simple_chemical
-	I-Simple_chemical
chloromethylthiochromone	I-Simple_chemical
-	I-Simple_chemical
1	I-Simple_chemical
,	I-Simple_chemical
1	I-Simple_chemical
-	I-Simple_chemical
dioxide	I-Simple_chemical
on	O
nucleic	B-Simple_chemical
acid	I-Simple_chemical
,	O
protein	O
,	O
and	O
aerobic	O
and	O
anaerobic	O
metabolism	O
of	O
Ehrlich	B-Cell
ascites	I-Cell
tumor	I-Cell
cells	I-Cell
.	O

3	B-Simple_chemical
-	I-Simple_chemical
Chloromethylthiochromone	I-Simple_chemical
-	I-Simple_chemical
1	I-Simple_chemical
,	I-Simple_chemical
1	I-Simple_chemical
-	I-Simple_chemical
dioxide	I-Simple_chemical
was	O
observed	O
to	O
be	O
a	O
potent	O
inhibitor	O
of	O
Ehrlich	B-Cancer
ascites	I-Cancer
carcinoma	I-Cancer
growth	O
and	O
a	O
moderate	O
inhibitor	O
of	O
P	B-Cancer
-	I-Cancer
388	I-Cancer
lymphocytic	I-Cancer
leukemia	I-Cancer
growth	O
at	O
10	O
mg	O
/	O
kg	O
/	O
day	O
.	O

Preliminary	O
in	O
vitro	O
studies	O
showed	O
that	O
the	O
agents	O
significantly	O
inhibited	O
RNA	B-Simple_chemical
and	O
DNA	B-Cellular_component
synthesis	O
in	O
Ehrlich	B-Cell
ascites	I-Cell
cells	I-Cell
.	O

In	O
vivo	O
studies	O
after	O
dosing	O
on	O
Days	O
6	O
,	O
7	O
,	O
and	O
8	O
demonstrated	O
the	O
same	O
reductions	O
in	O
nucleic	B-Simple_chemical
acid	I-Simple_chemical
synthesis	O
and	O
a	O
moderate	O
reduction	O
in	O
protein	O
synthesis	O
.	O

The	O
primary	O
site	O
of	O
nucleic	B-Simple_chemical
acid	I-Simple_chemical
synthesis	O
,	O
which	O
was	O
blocked	O
by	O
3	B-Simple_chemical
-	I-Simple_chemical
chloromethylthiochromone	I-Simple_chemical
,	O
was	O
at	O
orotidine	B-Gene_or_gene_product
monophosphate	I-Gene_or_gene_product
decarboxylase	I-Gene_or_gene_product
in	O
the	O
primidine	B-Simple_chemical
pathway	O
.	O

Other	O
enzymes	O
,	O
in	O
anaerobic	O
and	O
aerobic	O
glycolysis	O
,	O
which	O
were	O
blocked	O
include	O
hexokinase	B-Gene_or_gene_product
,	O
phosphofructokinase	B-Gene_or_gene_product
,	O
succinic	B-Gene_or_gene_product
and	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ketoglutarate	I-Gene_or_gene_product
dehydrogenases	I-Gene_or_gene_product
,	O
as	O
well	O
as	O
States	O
3	O
and	O
4	O
of	O
oxidative	O
phosphorylation	O
.	O

Mast	B-Cell
cell	I-Cell
heparin	B-Simple_chemical
stimulates	O
migration	O
of	O
capillary	B-Cell
endothelial	I-Cell
cells	I-Cell
in	O
vitro	O
.	O

Migration	O
of	O
capillary	B-Cell
endothelial	I-Cell
cells	I-Cell
is	O
an	O
important	O
component	O
of	O
angiogenesis	O
in	O
vivo	O
.	O

Increased	O
numbers	O
of	O
mast	B-Cell
cells	I-Cell
have	O
been	O
associated	O
with	O
several	O
types	O
of	O
angiogenesis	O
.	O

We	O
have	O
used	O
a	O
quantitative	O
assay	O
in	O
vitro	O
to	O
demonstrate	O
that	O
mast	B-Cell
cells	I-Cell
release	O
a	O
factor	O
that	O
significantly	O
increases	O
bovine	B-Organism
capillary	B-Cell
endothelial	I-Cell
cell	I-Cell
migration	O
.	O

The	O
factor	O
is	O
present	O
in	O
medium	O
conditioned	O
by	O
mast	B-Cell
cells	I-Cell
as	O
well	O
as	O
lysates	O
of	O
mast	B-Cell
cells	I-Cell
.	O

The	O
stimulatory	O
effect	O
of	O
mast	B-Cell
cells	I-Cell
on	O
migration	O
is	O
specific	O
for	O
capillary	B-Cell
endothelial	I-Cell
cells	I-Cell
.	O

Furthermore	O
,	O
mast	B-Cell
cells	I-Cell
have	O
no	O
mitogenic	O
activity	O
for	O
capillary	B-Cell
endothelial	I-Cell
cells	I-Cell
.	O

Of	O
all	O
the	O
secretory	O
products	O
of	O
mast	B-Cell
cells	I-Cell
tested	O
,	O
only	O
heparin	B-Simple_chemical
stimulated	O
capillary	B-Cell
endothelial	I-Cell
cell	I-Cell
migration	O
in	O
vitro	O
.	O

Heparin	B-Simple_chemical
preparations	O
from	O
a	O
variety	O
of	O
sources	O
stimulated	O
capillary	B-Cell
endothelial	I-Cell
cell	I-Cell
migration	O
to	O
the	O
same	O
degree	O
but	O
did	O
not	O
stimulate	O
migration	O
of	O
several	O
other	O
cell	B-Cell
types	O
.	O

The	O
migration	O
activity	O
of	O
heparin	B-Simple_chemical
and	O
mast	B-Cell
cell	I-Cell
conditioned	O
medium	O
was	O
blocked	O
by	O
specific	O
antagonists	O
of	O
heparin	B-Simple_chemical
(	O
protamine	B-Gene_or_gene_product
and	O
heparinase	B-Gene_or_gene_product
)	O
,	O
but	O
not	O
by	O
chondroitinase	B-Gene_or_gene_product
ABC	I-Gene_or_gene_product
.	O

The	O
migration	O
activity	O
of	O
mast	B-Cell
cell	I-Cell
conditioned	O
medium	O
was	O
resistant	O
to	O
heat	O
(	O
100	O
degrees	O
C	O
)	O
and	O
incubation	O
with	O
proteolytic	O
enzymes	O
.	O

These	O
results	O
suggest	O
that	O
the	O
role	O
of	O
mast	B-Cell
cells	I-Cell
in	O
angiogenesis	O
may	O
be	O
to	O
enhance	O
migration	O
of	O
the	O
endothelial	B-Cell
cells	I-Cell
of	O
growing	O
capillaries	B-Tissue
.	O

Cancer	B-Cancer
progression	O
and	O
p53	B-Gene_or_gene_product
.	O

In	O
a	O
complex	O
organism	O
,	O
somatic	B-Cell
cells	I-Cell
are	O
under	O
intermittent	O
selection	O
pressure	O
for	O
the	O
emergence	O
of	O
mutants	O
that	O
can	O
survive	O
environmental	O
insults	O
and	O
that	O
can	O
grow	O
autonomously	O
despite	O
adverse	O
conditions	O
.	O

Repeated	O
rounds	O
of	O
mutation	O
,	O
selection	O
,	O
and	O
proliferation	O
may	O
lead	O
to	O
cancer	B-Cancer
.	O

The	O
organism	O
prevents	O
malignant	O
transformation	O
by	O
assuring	O
accurate	O
DNA	B-Cellular_component
repair	O
before	O
cell	B-Cell
division	O
,	O
by	O
forcing	O
the	O
death	O
of	O
cells	B-Cell
with	O
excessive	O
DNA	B-Cellular_component
damage	O
,	O
and	O
by	O
placing	O
limits	O
on	O
the	O
replicative	O
lifespans	O
of	O
most	O
somatic	B-Cell
cells	I-Cell
.	O

The	O
p53	B-Gene_or_gene_product
gene	O
is	O
a	O
"	O
guardian	O
of	O
the	O
genome	O
"	O
-	O
-	O
it	O
regulates	O
multiple	O
components	O
of	O
the	O
DNA	B-Cellular_component
damage	O
control	O
response	O
and	O
promotes	O
cellular	B-Cell
senescence	O
.	O

Disabling	O
mutations	O
and	O
deletions	O
of	O
p53	B-Gene_or_gene_product
occur	O
in	O
50	O
%	O
of	O
human	B-Organism
tumours	B-Cancer
.	O

p53	B-Gene_or_gene_product
-	O
deficient	O
cancers	O
are	O
often	O
unstable	O
,	O
aggressive	O
,	O
and	O
resistant	O
to	O
therapy	O
.	O

The	O
Met	B-Gene_or_gene_product
proto	O
-	O
oncogene	O
mesenchymal	B-Cell
to	O
epithelial	B-Cell
cell	I-Cell
conversion	O
.	O

Coexpression	O
of	O
the	O
human	B-Organism
Met	B-Gene_or_gene_product
receptor	O
and	O
its	O
ligand	O
,	O
hepatocyte	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
/	O
scatter	B-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
HGF	B-Gene_or_gene_product
/	O
SF	B-Gene_or_gene_product
)	O
,	O
in	O
NIH	B-Cell
3T3	I-Cell
fibroblasts	I-Cell
causes	O
the	O
cells	B-Cell
to	O
become	O
tumorigenic	O
in	O
nude	B-Organism
mice	I-Organism
.	O

The	O
resultant	O
tumors	B-Cancer
display	O
lumen	B-Immaterial_anatomical_entity
-	O
like	O
morphology	O
,	O
contain	O
carcinoma	B-Cancer
-	O
like	O
focal	B-Cellular_component
areas	I-Cellular_component
with	O
intercellular	B-Cellular_component
junctions	I-Cellular_component
resembling	O
desmosomes	B-Cellular_component
,	O
and	O
coexpress	O
epithelial	B-Cell
(	O
cytokeratin	B-Gene_or_gene_product
)	O
and	O
mesenchymal	B-Cell
(	O
vimentin	B-Gene_or_gene_product
)	O
cytoskeletal	B-Cellular_component
markers	O
.	O

The	O
tumor	B-Cell
cells	I-Cell
also	O
display	O
enhanced	O
expression	O
of	O
desmosomal	B-Cellular_component
and	O
tight	B-Cellular_component
-	I-Cellular_component
junction	I-Cellular_component
proteins	O
.	O

The	O
apparent	O
mesenchymal	B-Cell
to	O
epithelial	B-Cell
conversion	O
of	O
the	O
tumor	B-Cell
cells	I-Cell
mimics	O
the	O
conversion	O
that	O
occurs	O
during	O
embryonic	B-Organ
kidney	I-Organ
development	O
,	O
suggesting	O
that	O
Met	B-Gene_or_gene_product
-	O
HGF	B-Gene_or_gene_product
/	O
SF	B-Gene_or_gene_product
signaling	O
plays	O
a	O
role	O
in	O
this	O
process	O
as	O
well	O
as	O
in	O
tumors	B-Cancer
that	O
express	O
both	O
epithelial	B-Cell
and	O
mesenchymal	B-Cell
markers	O
.	O

Ultraviolet	O
B	O
irradiation	O
promotes	O
tumorigenic	O
and	O
metastatic	O
properties	O
in	O
primary	O
cutaneous	B-Cancer
melanoma	I-Cancer
via	O
induction	O
of	O
interleukin	B-Gene_or_gene_product
8	I-Gene_or_gene_product
.	O

UV	O
radiation	O
has	O
been	O
shown	O
to	O
play	O
a	O
role	O
in	O
the	O
initiation	O
of	O
human	B-Organism
cutaneous	B-Cancer
melanoma	I-Cancer
,	O
but	O
its	O
role	O
in	O
the	O
development	O
of	O
malignant	B-Cancer
melanoma	I-Cancer
to	O
the	O
metastatic	O
state	O
is	O
not	O
very	O
well	O
defined	O
.	O

Although	O
previous	O
studies	O
have	O
concentrated	O
on	O
the	O
effect	O
of	O
UV	O
-	O
B	O
on	O
the	O
host	O
immune	O
response	O
,	O
the	O
effect	O
of	O
UV	O
-	O
B	O
on	O
the	O
tumor	B-Cell
cells	I-Cell
was	O
not	O
elucidated	O
.	O

Here	O
we	O
show	O
that	O
UV	O
-	O
B	O
can	O
induce	O
interleukin	B-Gene_or_gene_product
8	I-Gene_or_gene_product
(	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
)	O
mRNA	O
and	O
protein	O
secretion	O
in	O
human	B-Organism
cutaneous	B-Cancer
melanoma	I-Cancer
with	O
negligible	O
expression	O
of	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
.	O

UV	O
-	O
B	O
-	O
induced	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
was	O
constitutively	O
expressed	O
60	O
days	O
after	O
irradiation	O
in	O
tumors	B-Cancer
implanted	O
in	O
mice	B-Organism
.	O

Induction	O
of	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
was	O
UV	O
-	O
B	O
dose	O
dependent	O
and	O
blocked	O
by	O
cyclohexamide	B-Simple_chemical
,	O
indicating	O
that	O
de	O
novo	O
protein	O
synthesis	O
is	O
required	O
for	O
its	O
expression	O
.	O

The	O
UV	O
-	O
irradiated	O
cells	B-Cell
demonstrated	O
enhanced	O
tumorigenicity	O
and	O
metastatic	O
potential	O
in	O
nude	B-Organism
mice	I-Organism
.	O

The	O
increase	O
in	O
tumorigenicity	O
and	O
metastatic	O
ability	O
could	O
be	O
explained	O
by	O
the	O
increase	O
in	O
Mr	O
72	O
,	O
000	O
type	B-Gene_or_gene_product
IV	I-Gene_or_gene_product
collagenase	I-Gene_or_gene_product
activity	O
and	O
angiogenesis	O
attributed	O
to	O
the	O
induction	O
of	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
8	I-Gene_or_gene_product
after	O
irradiation	O
.	O

The	O
acquisition	O
of	O
the	O
metastatic	O
phenotype	O
induced	O
by	O
UV	O
-	O
B	O
could	O
not	O
be	O
attributed	O
to	O
abnormalities	O
in	O
the	O
p53	B-Gene_or_gene_product
or	O
MTS	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
(	O
p16INK4	B-Gene_or_gene_product
)	O
genes	O
.	O

To	O
the	O
best	O
of	O
our	O
knowledge	O
,	O
this	O
is	O
the	O
first	O
report	O
to	O
show	O
that	O
UV	O
-	O
B	O
can	O
increase	O
the	O
aggressiveness	O
of	O
human	B-Organism
cutaneous	B-Cancer
melanoma	I-Cancer
for	O
growth	O
and	O
metastasis	O
.	O

Cancer	B-Cancer
metastasis	O
:	O
negative	O
regulation	O
by	O
an	O
invasion	O
-	O
suppressor	O
complex	O
.	O

Invasion	O
is	O
the	O
hallmark	O
of	O
tumor	B-Cancer
malignancy	O
.	O

We	O
situate	O
invasion	O
within	O
microecosystems	O
comprising	O
neoplastic	B-Cell
cells	I-Cell
as	O
well	O
as	O
host	O
cells	B-Cell
.	O

Modulation	O
of	O
invasion	O
within	O
such	O
systems	O
is	O
ascribed	O
to	O
balances	O
between	O
promoter	O
and	O
suppressor	O
pathways	O
.	O

The	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
/	O
alpha	B-Gene_or_gene_product
-	I-Gene_or_gene_product
,	O
beta	B-Gene_or_gene_product
-	I-Gene_or_gene_product
,	O
gamma	B-Gene_or_gene_product
-	I-Gene_or_gene_product
catenin	I-Gene_or_gene_product
complex	O
has	O
an	O
invasion	O
-	O
suppressor	O
function	O
as	O
evidenced	O
by	O
transfections	O
either	O
with	O
sense	O
cDNA	O
encoding	O
these	O
molecules	O
or	O
with	O
antisense	O
cDNA	O
inhibiting	O
their	O
expression	O
.	O

Loss	O
of	O
heterozygosity	O
at	O
the	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Cadherin	I-Gene_or_gene_product
(	O
uvo	O
)	O
locus	O
has	O
been	O
reported	O
,	O
but	O
mutations	O
in	O
the	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
gene	O
seem	O
to	O
be	O
rare	O
.	O

Downregulation	O
of	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
occurred	O
at	O
the	O
level	O
of	O
transcription	O
or	O
of	O
mRNA	O
stability	O
.	O

Ex	O
vivo	O
cultures	B-Cell
from	O
invasive	B-Cancer
tumors	I-Cancer
or	O
metastases	O
produced	O
cells	B-Cell
that	O
were	O
homogeneously	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
-	O
positive	O
and	O
noninvasive	O
in	O
vitro	O
.	O

These	O
observations	O
have	O
led	O
to	O
the	O
idea	O
that	O
factors	O
in	O
the	O
host	O
downmodulate	O
the	O
E	B-Gene_or_gene_product
-	I-Gene_or_gene_product
cadherin	I-Gene_or_gene_product
complex	O
and	O
promote	O
invasion	O
most	O
probably	O
in	O
a	O
transient	O
way	O
.	O

Heparin	B-Simple_chemical
-	O
steroid	B-Simple_chemical
conjugates	O
:	O
new	O
angiogenesis	O
inhibitors	O
with	O
antitumor	B-Cancer
activity	O
in	O
mice	B-Organism
.	O

Inhibitors	O
of	O
angiogenesis	O
hold	O
potential	O
in	O
the	O
treatment	O
of	O
cancer	B-Cancer
and	O
other	O
diseases	O
where	O
the	O
disease	O
is	O
caused	O
or	O
maintained	O
by	O
the	O
inappropriate	O
growth	O
of	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
.	O

In	O
the	O
present	O
study	O
,	O
a	O
novel	O
inhibitor	O
of	O
angiogenesis	O
was	O
synthesized	O
by	O
covalently	O
linking	O
a	O
nonanticoagulating	O
derivative	O
of	O
heparin	B-Simple_chemical
,	O
heparin	B-Simple_chemical
adipic	I-Simple_chemical
hydrazide	I-Simple_chemical
(	O
HAH	B-Simple_chemical
)	O
,	O
by	O
an	O
acid	O
-	O
labile	O
bond	O
to	O
the	O
antiangiogenic	O
steroid	B-Simple_chemical
,	O
cortisol	B-Simple_chemical
.	O

The	O
rationale	O
was	O
that	O
the	O
heparin	B-Simple_chemical
derivative	O
,	O
which	O
binds	O
to	O
sulfated	O
polyanion	O
receptors	O
on	O
endothelial	B-Cell
cells	I-Cell
,	O
should	O
concentrate	O
the	O
steroid	B-Simple_chemical
on	O
the	O
surface	O
of	O
vascular	B-Cell
endothelial	I-Cell
cells	I-Cell
.	O

Endocytosis	O
of	O
the	O
conjugate	O
and	O
decomposition	O
of	O
the	O
acid	O
-	O
labile	O
linkage	O
inside	O
lysosomes	B-Cellular_component
and	O
other	O
acidic	O
intracellular	B-Cellular_component
compartments	I-Cellular_component
should	O
then	O
lead	O
to	O
release	O
of	O
the	O
cortisol	B-Simple_chemical
and	O
expression	O
of	O
its	O
antiproliferative	O
activity	O
.	O

Analysis	O
of	O
the	O
stability	O
of	O
HAH	B-Simple_chemical
-	I-Simple_chemical
cortisol	I-Simple_chemical
showed	O
that	O
it	O
was	O
stable	O
at	O
pH	O
7	O
.	O
4	O
and	O
broke	O
down	O
rapidly	O
(	O
t1	O
/	O
2	O
15	O
min	O
)	O
at	O
pH	O
4	O
.	O
8	O
at	O
37	O
degrees	O
C	O
.	O

Treatment	O
of	O
murine	B-Organism
pulmonary	B-Cell
capillary	I-Cell
endothelial	I-Cell
cells	I-Cell
with	O
HAH	B-Simple_chemical
-	I-Simple_chemical
cortisol	I-Simple_chemical
at	O
10	O
(	O
-	O
5	O
)	O
M	O
(	O
with	O
respect	O
to	O
cortisol	B-Simple_chemical
)	O
suppressed	O
their	O
DNA	B-Cellular_component
synthesis	O
by	O
50	O
%	O
and	O
inhibited	O
their	O
migration	O
into	O
wounded	B-Pathological_formation
areas	I-Pathological_formation
of	O
confluent	O
monolayers	B-Cell
.	O

HAH	B-Simple_chemical
-	I-Simple_chemical
cortisol	I-Simple_chemical
at	O
10	O
(	O
-	O
4	O
)	O
M	O
(	O
with	O
respect	O
to	O
cortisol	B-Simple_chemical
)	O
did	O
not	O
suppress	O
the	O
DNA	B-Cellular_component
synthesis	O
of	O
Lewis	B-Cell
lung	I-Cell
carcinoma	I-Cell
cells	I-Cell
.	O

Daily	O
i	O
.	O
p	O
.	O
injections	O
of	O
HAH	B-Simple_chemical
-	I-Simple_chemical
cortisol	I-Simple_chemical
into	O
mice	B-Organism
bearing	O
s	O
.	O
c	O
.	O
sponge	O
implants	O
retarded	O
vascularization	O
of	O
the	O
sponge	O
,	O
and	O
injections	O
directly	O
into	O
the	O
sponge	O
abolished	O
vascularization	O
for	O
as	O
long	O
as	O
the	O
injections	O
were	O
continued	O
.	O

Daily	O
i	B-Immaterial_anatomical_entity
.	I-Immaterial_anatomical_entity
v	I-Immaterial_anatomical_entity
.	I-Immaterial_anatomical_entity
injections	O
of	O
HAH	B-Simple_chemical
-	I-Simple_chemical
cortisol	I-Simple_chemical
at	O
doses	O
causing	O
no	O
apparent	O
toxicity	O
retarded	O
the	O
growth	O
of	O
solid	O
s	O
.	O
c	O
.	O

Lewis	B-Cancer
lung	I-Cancer
carcinomas	I-Cancer
in	O
mice	B-Organism
by	O
up	O
to	O
65	O
%	O
.	O

In	O
all	O
of	O
these	O
assays	O
,	O
equivalent	O
treatments	O
with	O
a	O
mixture	O
of	O
the	O
HAH	B-Simple_chemical
plus	O
cortisol	B-Simple_chemical
was	O
significantly	O
less	O
effective	O
.	O

The	O
antiproliferative	O
effect	O
of	O
HAH	B-Simple_chemical
-	I-Simple_chemical
cortisol	I-Simple_chemical
on	O
endothelial	B-Cell
cells	I-Cell
appeared	O
independent	O
of	O
the	O
glucocorticoid	B-Simple_chemical
activity	O
of	O
the	O
steroid	B-Simple_chemical
since	O
HAH	B-Simple_chemical
conjugated	O
to	O
5	B-Simple_chemical
beta	I-Simple_chemical
-	I-Simple_chemical
pregnane	I-Simple_chemical
-	I-Simple_chemical
3	I-Simple_chemical
alpha	I-Simple_chemical
,	I-Simple_chemical
17	I-Simple_chemical
alpha	I-Simple_chemical
,	I-Simple_chemical
21	I-Simple_chemical
-	I-Simple_chemical
triol	I-Simple_chemical
-	I-Simple_chemical
20	I-Simple_chemical
-	I-Simple_chemical
one	I-Simple_chemical
,	O
a	O
steroid	B-Simple_chemical
lacking	O
glucocorticoid	B-Simple_chemical
or	O
mineralocorticoid	B-Simple_chemical
activity	O
,	O
was	O
even	O
more	O
effective	O
at	O
inhibiting	O
DNA	B-Cellular_component
synthesis	O
by	O
murine	B-Organism
pulmonary	B-Cell
capillary	I-Cell
endothelial	I-Cell
cells	I-Cell
than	O
was	O
HAH	B-Simple_chemical
-	I-Simple_chemical
cortisol	I-Simple_chemical
.	O

In	O
conclusion	O
,	O
HAH	B-Simple_chemical
-	I-Simple_chemical
cortisol	I-Simple_chemical
represents	O
the	O
prototype	O
of	O
a	O
new	O
class	O
of	O
angiogenesis	O
inhibitors	O
for	O
the	O
treatment	O
of	O
cancer	B-Cancer
and	O
other	O
angiogenic	O
diseases	O
.	O

Selenoperoxidase	B-Gene_or_gene_product
-	O
dependent	O
glutathione	B-Simple_chemical
cycle	O
activity	O
in	O
peroxide	B-Simple_chemical
-	O
challenged	O
leukemia	B-Cell
cells	I-Cell
.	O

Murine	B-Organism
leukemia	B-Cell
L1210	I-Cell
cells	I-Cell
rendered	O
deficient	O
in	O
glutathione	B-Gene_or_gene_product
peroxidase	I-Gene_or_gene_product
(	O
GPX	B-Gene_or_gene_product
)	O
and	O
phospholipid	B-Gene_or_gene_product
hydroperoxide	I-Gene_or_gene_product
glutathione	I-Gene_or_gene_product
peroxidase	I-Gene_or_gene_product
(	O
PHGPX	B-Gene_or_gene_product
)	O
by	O
Se	B-Simple_chemical
deprivation	O
(	O
L	B-Cell
.	I-Cell
Se	I-Cell
(	I-Cell
-	I-Cell
)	I-Cell
cells	I-Cell
)	O
were	O
found	O
to	O
be	O
more	O
sensitive	O
to	O
tert	B-Simple_chemical
-	I-Simple_chemical
butyl	I-Simple_chemical
hydroperoxide	I-Simple_chemical
(	O
t	B-Simple_chemical
-	I-Simple_chemical
BuOOH	I-Simple_chemical
)	O
cytotoxicity	O
than	O
Se	B-Simple_chemical
-	O
replete	O
controls	O
(	O
L	B-Cell
.	I-Cell
Se	I-Cell
(	I-Cell
+	I-Cell
)	I-Cell
cells	I-Cell
)	O
.	O

Human	B-Organism
K562	B-Cell
cells	I-Cell
,	O
which	O
express	O
PHGPX	B-Gene_or_gene_product
,	O
but	O
not	O
GPX	B-Gene_or_gene_product
,	O
were	O
also	O
more	O
sensitive	O
to	O
t	B-Simple_chemical
-	I-Simple_chemical
BuOOH	I-Simple_chemical
in	O
the	O
Se	B-Simple_chemical
-	O
deficient	O
(	O
K	B-Cell
.	I-Cell
Se	I-Cell
(	I-Cell
-	I-Cell
)	I-Cell
)	O
than	O
Se	B-Simple_chemical
-	O
satisfied	O
(	O
K	B-Cell
.	I-Cell
Se	I-Cell
(	I-Cell
+	I-Cell
)	I-Cell
)	O
condition	O
.	O

In	O
examining	O
the	O
metabolic	O
basis	O
for	O
selenoperoxidase	B-Gene_or_gene_product
-	O
dependent	O
resistance	O
,	O
we	O
found	O
that	O
glucose	B-Simple_chemical
-	O
replete	O
Se	B-Cell
(	I-Cell
-	I-Cell
)	I-Cell
cells	I-Cell
reduce	O
t	B-Simple_chemical
-	I-Simple_chemical
BuOOH	I-Simple_chemical
to	O
t	B-Simple_chemical
-	I-Simple_chemical
butanol	I-Simple_chemical
far	O
more	O
slowly	O
than	O
Se	B-Cell
(	I-Cell
+	I-Cell
)	I-Cell
cells	I-Cell
,	O
the	O
ratio	O
of	O
the	O
first	O
-	O
order	O
rate	O
constants	O
approximating	O
that	O
of	O
the	O
GPX	B-Gene_or_gene_product
activities	O
(	O
L1210	B-Cell
cells	I-Cell
)	O
or	O
PHGPX	B-Gene_or_gene_product
activities	O
(	O
K562	B-Cell
cells	I-Cell
)	O
.	O

Monitoring	O
peroxide	B-Simple_chemical
-	O
induced	O
changes	O
in	O
GSH	B-Simple_chemical
and	O
GSSG	B-Simple_chemical
gave	O
consistent	O
results	O
;	O
e	O
.	O
g	O
.	O
,	O
glucose	B-Simple_chemical
-	O
depleted	O
L	O
.	O
Se	B-Cell
(	I-Cell
+	I-Cell
)	I-Cell
cells	I-Cell
exhibited	O
a	O
first	O
order	O
loss	O
of	O
GSH	B-Simple_chemical
that	O
was	O
substantially	O
faster	O
than	O
that	O
of	O
glucose	B-Simple_chemical
-	O
depleted	O
L	B-Cell
.	I-Cell
Se	I-Cell
(	I-Cell
-	I-Cell
)	I-Cell
cells	I-Cell
.	O

Under	O
the	O
conditions	O
used	O
,	O
peroxide	B-Simple_chemical
-	O
induced	O
conversion	O
of	O
GSH	B-Simple_chemical
to	O
GSSG	B-Simple_chemical
could	O
be	O
stoichiometrically	O
reversed	O
by	O
resupplying	O
D	B-Simple_chemical
-	I-Simple_chemical
glucose	I-Simple_chemical
,	O
indicating	O
that	O
no	O
significant	O
lysis	O
or	O
GSSG	B-Simple_chemical
efflux	O
and	O
/	O
or	O
interchange	O
had	O
taken	O
place	O
.	O

The	O
apparent	O
first	O
-	O
order	O
rate	O
constant	O
for	O
GSH	B-Simple_chemical
decay	O
increased	O
progressively	O
for	O
L1210	B-Cell
cells	I-Cell
expressing	O
a	O
range	O
of	O
GPX	B-Gene_or_gene_product
activities	O
from	O
approximately	O
5	O
%	O
to	O
100	O
%	O
,	O
demonstrating	O
that	O
peroxide	B-Simple_chemical
detoxification	O
is	O
strictly	O
dependent	O
on	O
enzyme	O
content	O
.	O

The	O
initial	O
rate	O
of	O
14CO2	B-Simple_chemical
release	O
from	O
D	B-Simple_chemical
-	I-Simple_chemical
[	I-Simple_chemical
1	I-Simple_chemical
-	I-Simple_chemical
14C	I-Simple_chemical
]	I-Simple_chemical
glucose	I-Simple_chemical
supplied	O
in	O
the	O
medium	O
was	O
much	O
greater	O
for	O
L	B-Cell
.	I-Cell
Se	I-Cell
(	I-Cell
+	I-Cell
)	I-Cell
or	O
K	B-Cell
.	I-Cell
Se	I-Cell
(	I-Cell
+	I-Cell
)	I-Cell
cells	I-Cell
than	O
for	O
their	O
respective	O
Se	B-Cell
(	I-Cell
-	I-Cell
)	I-Cell
counterparts	O
,	O
consistent	O
with	O
greater	O
hexose	B-Simple_chemical
monophosphate	I-Simple_chemical
shunt	O
activity	O
in	O
the	O
former	O
.	O

These	O
results	O
highlight	O
the	O
importance	O
of	O
selenoperoxidase	B-Gene_or_gene_product
action	O
in	O
the	O
glutathione	B-Simple_chemical
cycle	O
as	O
a	O
means	O
by	O
which	O
tumor	B-Cell
cells	I-Cell
cope	O
with	O
hydroperoxide	B-Simple_chemical
stress	O
.	O

Differential	O
expression	O
and	O
mutation	O
of	O
NME	B-Gene_or_gene_product
genes	O
in	O
autologous	O
cultured	O
human	B-Organism
melanoma	B-Cell
cells	I-Cell
with	O
different	O
metastatic	O
potentials	O
.	O

The	O
putative	O
metastasis	O
suppressor	O
genes	O
,	O
NME1	B-Gene_or_gene_product
(	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
and	O
NME2	B-Gene_or_gene_product
(	O
nm23	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
)	O
,	O
were	O
examined	O
in	O
a	O
model	O
system	O
we	O
developed	O
to	O
approximate	O
the	O
dissemination	O
of	O
melanoma	B-Cancer
from	O
a	O
primary	O
skin	B-Cancer
tumor	I-Cancer
.	O

We	O
utilized	O
two	O
autologous	O
human	B-Organism
melanoma	B-Cell
cell	I-Cell
lines	I-Cell
,	O
IV	B-Cell
Cl	I-Cell
1	I-Cell
and	O
IV	B-Cell
Cl	I-Cell
3	I-Cell
,	O
which	O
displayed	O
qualitatively	O
different	O
metastatic	O
phenotypes	O
following	O
subdermal	B-Organism_subdivision
inoculation	O
into	O
nude	B-Organism
mice	I-Organism
.	O

Highly	O
metastatic	O
IV	B-Cell
Cl	I-Cell
1	I-Cell
cells	I-Cell
expressed	O
approximately	O
5	O
fold	O
lower	O
levels	O
of	O
protein	O
encoded	O
by	O
NME	B-Gene_or_gene_product
genes	O
than	O
non	O
-	O
metastatic	O
IV	B-Cell
Cl	I-Cell
3	I-Cell
cells	I-Cell
.	O

Similar	O
differences	O
in	O
NME	B-Gene_or_gene_product
protein	O
levels	O
were	O
observed	O
in	O
tumors	B-Cancer
induced	O
by	O
the	O
two	O
cell	B-Cell
lines	I-Cell
in	O
nude	B-Organism
mice	I-Organism
.	O

There	O
were	O
no	O
differences	O
in	O
NME	B-Gene_or_gene_product
mRNA	O
levels	O
between	O
these	O
two	O
cell	B-Cell
lines	I-Cell
,	O
suggesting	O
that	O
expression	O
of	O
these	O
proteins	O
is	O
regulated	O
at	O
a	O
post	O
-	O
transcriptional	O
level	O
.	O

We	O
found	O
a	O
ser122	B-Amino_acid
-	O
pro	B-Amino_acid
mutation	O
in	O
the	O
NME2	B-Gene_or_gene_product
gene	O
of	O
metastatic	O
IV	B-Cell
Cl	I-Cell
1	I-Cell
cells	I-Cell
.	O

A	O
similar	O
ser120	B-Amino_acid
-	O
gly	B-Amino_acid
mutation	O
in	O
NME1	B-Gene_or_gene_product
has	O
been	O
found	O
in	O
human	B-Organism
neuroblastoma	B-Cancer
,	O
suggesting	O
that	O
mutation	O
in	O
this	O
region	O
may	O
be	O
a	O
general	O
phenomenon	O
related	O
to	O
tumor	B-Cancer
progression	O
.	O

These	O
mutations	O
may	O
have	O
functional	O
consequences	O
since	O
they	O
eliminate	O
potential	O
phosphorylation	O
sites	O
and	O
may	O
affect	O
the	O
tertiary	O
structure	O
of	O
mature	O
protein	O
complexes	O
.	O

Cyclin	B-Gene_or_gene_product
D1	I-Gene_or_gene_product
/	O
bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
cooperates	O
with	O
myc	B-Gene_or_gene_product
genes	O
in	O
the	O
generation	O
of	O
B	B-Cancer
-	I-Cancer
cell	I-Cancer
lymphoma	I-Cancer
in	O
transgenic	B-Organism
mice	I-Organism
.	O

The	O
chromosomal	B-Cellular_component
translocation	O
t	O
(	O
11	O
:	O
14	O
)	O
is	O
associated	O
with	O
human	B-Organism
lymphoid	B-Pathological_formation
neoplasia	I-Pathological_formation
affecting	O
centrocytic	B-Cell
B	I-Cell
-	I-Cell
cells	I-Cell
of	O
intermediate	O
differentiation	O
.	O

As	O
a	O
consequence	O
the	O
cyclin	B-Gene_or_gene_product
D1	I-Gene_or_gene_product
(	O
bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
)	O
gene	O
is	O
juxtaposed	O
to	O
the	O
immunoglobulin	B-Gene_or_gene_product
heavy	I-Gene_or_gene_product
chain	I-Gene_or_gene_product
enhancer	I-Gene_or_gene_product
E	I-Gene_or_gene_product
mu	I-Gene_or_gene_product
.	O

To	O
show	O
that	O
transcriptional	O
activation	O
of	O
cyclin	B-Gene_or_gene_product
D1	I-Gene_or_gene_product
is	O
causally	O
involved	O
in	O
the	O
generation	O
of	O
B	B-Cancer
-	I-Cancer
cell	I-Cancer
neoplasia	I-Cancer
we	O
have	O
generated	O
transgenic	B-Organism
mice	I-Organism
that	O
carry	O
a	O
cyclin	B-Gene_or_gene_product
D1	I-Gene_or_gene_product
gene	O
under	O
the	O
transcriptional	O
control	O
of	O
the	O
E	B-Gene_or_gene_product
mu	I-Gene_or_gene_product
element	O
.	O

E	B-Gene_or_gene_product
mu	I-Gene_or_gene_product
cyclin	B-Gene_or_gene_product
D1	I-Gene_or_gene_product
transgenic	O
mice	O
show	O
only	O
very	O
subtle	O
alterations	O
in	O
the	O
cycling	O
behaviour	O
of	O
B	B-Cell
-	I-Cell
cell	I-Cell
populations	I-Cell
in	O
the	O
bone	B-Multi-tissue_structure
marrow	I-Multi-tissue_structure
compared	O
with	O
normal	O
mice	B-Organism
and	O
do	O
not	O
develop	O
lymphoid	B-Cancer
tumours	I-Cancer
.	O

However	O
,	O
E	B-Gene_or_gene_product
mu	I-Gene_or_gene_product
-	O
directed	O
coexpression	O
of	O
cyclin	B-Gene_or_gene_product
D1	I-Gene_or_gene_product
and	O
N	B-Gene_or_gene_product
-	I-Gene_or_gene_product
MYC	I-Gene_or_gene_product
or	O
L	B-Gene_or_gene_product
-	I-Gene_or_gene_product
MYC	I-Gene_or_gene_product
in	O
double	O
transgenic	B-Organism
mice	I-Organism
reveals	O
a	O
strong	O
cooperative	O
effect	O
between	O
MYC	B-Gene_or_gene_product
and	O
cyclin	B-Gene_or_gene_product
D1	I-Gene_or_gene_product
provoking	O
the	O
rapid	O
development	O
of	O
clonal	O
pre	B-Cancer
-	I-Cancer
B	I-Cancer
and	O
B	B-Cancer
-	I-Cancer
cell	I-Cancer
lymphomas	I-Cancer
.	O

Interestingly	O
,	O
crossing	O
of	O
cyclin	B-Gene_or_gene_product
D1	I-Gene_or_gene_product
transgenic	O
mice	O
with	O
E	B-Gene_or_gene_product
mu	I-Gene_or_gene_product
L	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
transgenics	O
that	O
express	O
their	O
transgene	O
in	O
both	O
B	B-Cell
-	I-Cell
and	O
T	B-Cell
-	I-Cell
cells	I-Cell
but	O
predominantly	O
develop	O
T	B-Cancer
-	I-Cancer
cell	I-Cancer
tumours	I-Cancer
leads	O
in	O
double	O
transgenics	B-Organism
exclusively	O
to	O
B	B-Cell
-	I-Cell
cell	I-Cell
neoplasia	I-Cell
.	O

The	O
data	O
presented	O
here	O
demonstrate	O
that	O
transcriptional	O
activation	O
of	O
cyclin	B-Gene_or_gene_product
D1	I-Gene_or_gene_product
can	O
oncogenically	O
transform	O
B	B-Cell
-	I-Cell
cells	I-Cell
in	O
concert	O
with	O
a	O
myc	B-Gene_or_gene_product
gene	O
.	O

They	O
establish	O
cyclin	B-Gene_or_gene_product
D1	I-Gene_or_gene_product
as	O
a	O
proto	O
-	O
oncogene	O
whose	O
activity	O
appears	O
to	O
depend	O
on	O
a	O
specific	O
cell	B-Cell
type	O
as	O
well	O
as	O
on	O
a	O
specific	O
cooperating	O
partner	O
and	O
link	O
disturbances	O
in	O
the	O
regulation	O
of	O
cell	B-Cell
cycle	O
progression	O
to	O
the	O
development	O
of	O
human	B-Organism
malignancies	B-Cancer
.	O

Elevated	O
expression	O
of	O
the	O
junB	B-Gene_or_gene_product
proto	O
-	O
oncogene	O
is	O
essential	O
for	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
fos	I-Gene_or_gene_product
induced	O
transformation	O
of	O
Rat	B-Cell
-	I-Cell
1	I-Cell
cells	I-Cell
.	O

We	O
previously	O
described	O
the	O
isolation	O
of	O
non	O
-	O
tumorigenic	O
revertants	O
from	O
mutagenized	O
populations	O
of	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
fos	I-Gene_or_gene_product
-	O
transformed	O
Rat	B-Cell
-	I-Cell
1	I-Cell
cells	I-Cell
(	O
Zarbl	O
et	O
al	O
.	O
,	O
1987	O
)	O
.	O

In	O
the	O
present	O
study	O
we	O
examined	O
the	O
possibility	O
that	O
the	O
revertant	B-Cell
phenotype	O
resulted	O
from	O
mutations	O
that	O
altered	O
the	O
expression	O
or	O
activities	O
of	O
the	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
jun	I-Gene_or_gene_product
or	O
junB	B-Gene_or_gene_product
proto	O
-	O
oncogenes	O
.	O

The	O
results	O
demonstrated	O
that	O
levels	O
of	O
the	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
jun	I-Gene_or_gene_product
mRNA	O
and	O
protein	O
were	O
unchanged	O
in	O
the	O
revertants	B-Cell
when	O
compared	O
to	O
the	O
transformed	O
parental	O
cells	B-Cell
,	O
and	O
ectopic	O
overexpression	O
of	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
jun	I-Gene_or_gene_product
failed	O
to	O
retransform	O
the	O
revertants	B-Cell
.	O

Although	O
one	O
mutant	O
allele	O
was	O
detected	O
in	O
revertant	B-Cell
EMS	I-Cell
-	I-Cell
1	I-Cell
-	I-Cell
19	I-Cell
,	O
overexpression	O
of	O
this	O
mutant	O
allele	O
failed	O
to	O
inhibit	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
fos	I-Gene_or_gene_product
induced	O
cell	B-Cell
transformation	O
.	O

Together	O
these	O
results	O
indicated	O
that	O
the	O
revertant	B-Cell
phenotype	O
did	O
not	O
result	O
from	O
altered	O
expression	O
or	O
mutations	O
in	O
the	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
jun	I-Gene_or_gene_product
gene	O
.	O

In	O
contrast	O
to	O
the	O
results	O
obtained	O
with	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
jun	I-Gene_or_gene_product
,	O
the	O
levels	O
of	O
junB	B-Gene_or_gene_product
mRNA	O
and	O
protein	O
were	O
found	O
to	O
be	O
reduced	O
two	O
-	O
or	O
threefold	O
in	O
revertant	B-Cell
EMS	I-Cell
-	I-Cell
1	I-Cell
-	I-Cell
19	I-Cell
.	O

Ectopic	O
overexpression	O
of	O
junB	B-Gene_or_gene_product
induced	O
transformation	O
of	O
revertant	O
EMS	B-Cell
-	I-Cell
1	I-Cell
-	I-Cell
19	I-Cell
,	O
but	O
failed	O
to	O
transform	O
Rat	B-Cell
-	I-Cell
1	I-Cell
cells	I-Cell
.	O

Moreover	O
,	O
about	O
10	O
%	O
of	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
fos	I-Gene_or_gene_product
transformed	O
cells	B-Cell
transfected	O
with	O
vectors	O
that	O
express	O
antisense	O
junB	B-Gene_or_gene_product
mRNA	O
acquired	O
a	O
non	O
-	O
transformed	O
phenotype	O
.	O

Together	O
these	O
results	O
indicate	O
that	O
expression	O
of	O
junB	B-Gene_or_gene_product
above	O
a	O
threshold	O
level	O
is	O
essential	O
for	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
fos	I-Gene_or_gene_product
-	O
induced	O
transformation	O
of	O
Rat	B-Cell
-	I-Cell
1	I-Cell
fibroblasts	I-Cell
.	O

[	O
Treatment	O
of	O
the	O
diabetic	O
foot	B-Organism_subdivision
by	O
hyperbaric	B-Simple_chemical
oxygen	I-Simple_chemical
]	O

Diabetic	O
foot	B-Pathological_formation
wounds	I-Pathological_formation
are	O
consequences	O
of	O
the	O
neuropathy	O
and	O
the	O
small	O
and	O
large	O
vessel	B-Multi-tissue_structure
disease	O
that	O
complicate	O
diabetes	O
.	O

At	O
the	O
cellular	B-Cell
level	O
,	O
the	O
result	O
is	O
hypoxia	O
which	O
impairs	O
wound	B-Pathological_formation
healing	O
.	O

Hyperbaric	O
oxygenation	O
(	O
HBO	O
)	O
may	O
be	O
a	O
useful	O
adjuvant	O
to	O
wound	B-Pathological_formation
care	O
.	O

It	O
leads	O
to	O
enhanced	O
oxygenation	O
of	O
the	O
affected	O
tissues	B-Tissue
,	O
has	O
an	O
antiseptic	O
effect	O
,	O
reduces	O
edema	B-Pathological_formation
,	O
and	O
accelerates	O
collagen	B-Gene_or_gene_product
production	O
and	O
angiogenesis	O
,	O
thus	O
enhancing	O
tissue	B-Tissue
repair	O
.	O

14	O
diabetics	O
with	O
chronic	B-Pathological_formation
nonhealing	I-Pathological_formation
wounds	I-Pathological_formation
which	O
did	O
not	O
respond	O
to	O
treatment	O
for	O
at	O
least	O
3	O
months	O
were	O
treated	O
by	O
HBO	O
.	O

All	O
had	O
palpable	O
pedal	O
pulses	O
.	O

Transcutaneous	O
measurements	O
of	O
tissue	B-Tissue
pO2	O
showed	O
elevation	O
from	O
20	O
+	O
/	O
-	O
10	O
mm	O
Hg	O
during	O
air	O
breathing	O
to	O
643	O
+	O
/	O
-	O
242	O
mm	O
Hg	O
while	O
breathing	O
pure	O
oxygen	B-Simple_chemical
at	O
2	O
.	O
5	O
ATA	O
.	O

They	O
were	O
treated	O
with	O
HBO	O
in	O
56	O
+	O
/	O
-	O
10	O
consecutive	O
HBO	O
sessions	O
.	O

In	O
11	O
there	O
was	O
complete	O
wound	B-Pathological_formation
healing	O
,	O
while	O
in	O
1	O
there	O
was	O
partial	O
response	O
,	O
in	O
1	O
minimal	O
response	O
,	O
and	O
in	O
1	O
a	O
transient	O
response	O
.	O

HBO	O
is	O
useful	O
in	O
chronic	B-Pathological_formation
nonhealing	I-Pathological_formation
wounds	I-Pathological_formation
of	O
the	O
diabetic	O
foot	B-Organism_subdivision
and	O
of	O
the	O
diabetic	O
foot	B-Organism_subdivision
with	O
impending	O
amputation	O
.	O

It	O
is	O
a	O
safe	O
mode	O
of	O
therapy	O
,	O
but	O
further	O
studies	O
are	O
required	O
to	O
establish	O
its	O
efficacy	O
and	O
to	O
ascertain	O
which	O
diabetic	O
patients	B-Organism
and	O
wounds	B-Pathological_formation
will	O
benefit	O
the	O
most	O
from	O
it	O
.	O

Appearance	O
of	O
tumorous	B-Cancer
phenotypes	O
in	O
goldfish	B-Organism
erythrophores	B-Cell
transfected	O
with	O
ras	B-Gene_or_gene_product
,	O
src	B-Gene_or_gene_product
,	O
and	O
myc	B-Gene_or_gene_product
oncogenes	O
and	O
spontaneous	O
differentiation	O
of	O
the	O
transformants	B-Cell
in	O
vitro	O
.	O

When	O
goldfish	B-Organism
erythrophores	B-Cell
isolated	O
from	O
the	O
skin	B-Organ
by	O
tissue	B-Tissue
digestion	O
and	O
centrifugation	O
in	O
a	O
Percoll	O
density	O
gradient	O
were	O
transfected	O
in	O
a	O
monolayer	B-Cell
-	O
culture	O
with	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Ha	I-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
or	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
src	I-Gene_or_gene_product
oncogene	O
either	O
singly	O
or	O
in	O
combination	O
with	O
v	B-Gene_or_gene_product
-	I-Gene_or_gene_product
myc	I-Gene_or_gene_product
by	O
means	O
of	O
calcium	B-Simple_chemical
phosphate	I-Simple_chemical
-	O
DNA	B-Cellular_component
co	O
-	O
precipitation	O
,	O
there	O
appeared	O
a	O
certain	O
number	O
of	O
transformants	B-Cell
manifesting	O
a	O
chromatoblast	B-Cell
-	O
like	O
profile	O
and	O
tumorous	B-Cancer
phenotypes	O
as	O
seen	O
in	O
the	O
capability	O
for	O
unlimited	O
growth	O
,	O
and	O
piling	O
-	O
up	O
in	O
a	O
monolayer	B-Cell
-	O
culture	O
or	O
colony	B-Cell
formation	O
in	O
semi	O
-	O
solid	O
soft	O
agar	O
.	O

After	O
successive	O
growth	O
in	O
vitro	O
for	O
longer	O
than	O
one	O
month	O
which	O
was	O
scarcely	O
observed	O
with	O
the	O
erythrophores	B-Cell
,	O
the	O
vast	O
majority	O
of	O
such	O
transformants	B-Cell
began	O
to	O
differentiate	O
into	O
erythrophores	B-Cell
and	O
ceased	O
proliferation	O
spontaneously	O
.	O

The	O
onset	O
of	O
their	O
differentiation	O
was	O
ascertained	O
by	O
the	O
deposition	O
of	O
marker	O
pteridine	B-Simple_chemical
pigments	O
.	O

None	O
of	O
the	O
transformants	B-Cell
differentiated	O
into	O
melanophores	B-Cell
or	O
iridophores	B-Cell
or	O
other	O
neural	B-Tissue
crest	I-Tissue
derivatives	O
as	O
seen	O
in	O
goldfish	B-Organism
erythrophoroma	B-Cell
cells	I-Cell
.	O

Little	O
difference	O
was	O
observed	O
in	O
their	O
transforming	O
efficiency	O
(	O
0	O
.	O
2	O
-	O
0	O
.	O
3	O
transformants	B-Cell
/	O
micrograms	O
DNA	B-Cellular_component
)	O
between	O
the	O
combinations	O
of	O
oncogenes	O
applied	O
but	O
a	O
tendency	O
was	O
noted	O
that	O
cells	B-Cell
transfected	O
with	O
ras	B-Gene_or_gene_product
or	O
src	B-Gene_or_gene_product
in	O
combination	O
with	O
myc	B-Gene_or_gene_product
developed	O
the	O
capacity	O
to	O
grow	O
for	O
a	O
longer	O
period	O
and	O
differentiated	O
at	O
a	O
later	O
stage	O
than	O
those	O
transfected	O
solely	O
with	O
ras	B-Gene_or_gene_product
or	O
src	B-Gene_or_gene_product
.	O

One	O
cell	B-Cell
line	I-Cell
(	O
ESM	B-Cell
-	I-Cell
1	I-Cell
)	O
derived	O
from	O
the	O
erythrophores	B-Cell
transfected	O
with	O
src	B-Gene_or_gene_product
/	O
myc	B-Gene_or_gene_product
grew	O
successively	O
over	O
nine	O
months	O
,	O
indicating	O
its	O
acquisition	O
of	O
immortality	O
.	O

The	O
expression	O
of	O
the	O
transfected	O
oncogenes	O
in	O
this	O
cell	B-Cell
line	I-Cell
was	O
examined	O
in	O
comparison	O
with	O
the	O
erythrophoroma	B-Cell
cells	I-Cell
by	O
Western	O
and	O
Northern	O
blot	O
analyses	O
.	O

Immune	O
consequences	O
of	O
burn	O
injury	O
.	O

The	O
purpose	O
of	O
the	O
immune	B-Anatomical_system
system	I-Anatomical_system
is	O
to	O
protect	O
cells	B-Cell
from	O
invasion	O
by	O
microorganisms	O
.	O

The	O
body	B-Organism_subdivision
has	O
three	O
equally	O
important	O
interactive	O
immune	B-Anatomical_system
defense	I-Anatomical_system
systems	I-Anatomical_system
,	O
all	O
of	O
which	O
are	O
profoundly	O
disrupted	O
with	O
major	O
burn	O
injury	O
.	O

The	O
immune	O
response	O
to	O
burn	O
injury	O
is	O
immediate	O
,	O
prolonged	O
,	O
and	O
severe	O
.	O

The	O
end	O
result	O
in	O
individuals	O
surviving	O
burn	O
shock	O
is	O
immunosuppression	O
,	O
with	O
increased	O
susceptibility	O
to	O
potentially	O
fatal	O
systemic	O
burn	B-Pathological_formation
wound	I-Pathological_formation
or	O
pulmonary	B-Organ
sepsis	O
.	O

Nursing	O
actions	O
to	O
support	O
the	O
humoral	O
and	O
cell	B-Cell
-	O
mediated	O
immune	B-Anatomical_system
system	I-Anatomical_system
of	O
the	O
burned	O
patient	B-Organism
include	O
providing	O
nutritional	O
support	O
to	O
maintain	O
serum	B-Organism_substance
protein	O
levels	O
at	O
optimal	O
levels	O
;	O
measures	O
to	O
decrease	O
edema	B-Pathological_formation
and	O
promote	O
angiogenesis	O
in	O
areas	O
of	O
partial	O
-	O
thickness	O
injury	O
;	O
meticulous	O
treatment	O
of	O
the	O
wound	B-Pathological_formation
to	O
prevent	O
infection	O
and	O
promote	O
healing	O
;	O
monitoring	O
of	O
antibiotic	O
use	O
;	O
conservative	O
use	O
of	O
invasive	O
techniques	O
,	O
including	O
intubation	O
and	O
vascular	B-Multi-tissue_structure
access	O
devices	O
;	O
maintenance	O
of	O
fluid	B-Organism_substance
and	O
electrolyte	O
balance	O
and	O
body	B-Organism_subdivision
temperature	O
;	O
and	O
energy	O
conservation	O
measures	O
.	O

Autocrine	O
angiogenic	O
vascular	B-Multi-tissue_structure
prosthesis	O
with	O
bone	B-Multi-tissue_structure
marrow	I-Multi-tissue_structure
transplantation	O
.	O

Synthetic	O
vascular	B-Multi-tissue_structure
prostheses	O
are	O
foreign	O
bodies	O
,	O
so	O
that	O
blood	B-Organism_substance
coagulation	O
can	O
occur	O
on	O
their	O
luminal	B-Multi-tissue_structure
surfaces	I-Multi-tissue_structure
,	O
causing	O
graft	B-Multi-tissue_structure
occlusion	O
very	O
frequently	O
in	O
prostheses	O
of	O
small	O
diameter	O
.	O

A	O
vascular	B-Multi-tissue_structure
prosthesis	O
needs	O
angiogenesis	O
for	O
endothelialization	O
of	O
the	O
luminal	B-Multi-tissue_structure
surface	I-Multi-tissue_structure
,	O
as	O
endothelial	B-Cell
cells	I-Cell
have	O
natural	O
and	O
permanent	O
antithrombogenic	O
properties	O
.	O

To	O
induce	O
capillary	B-Tissue
growth	O
into	O
the	O
graft	O
,	O
we	O
developed	O
a	O
method	O
of	O
transplanting	O
bone	B-Cell
marrow	I-Cell
cells	I-Cell
,	O
which	O
are	O
primitive	O
,	O
strong	O
enough	O
to	O
survive	O
,	O
and	O
create	O
blood	B-Cell
cells	I-Cell
,	O
resulting	O
in	O
the	O
inducement	O
of	O
capillary	B-Tissue
growth	O
.	O

In	O
an	O
animal	O
experiment	O
,	O
marrow	B-Cell
cells	I-Cell
were	O
infiltrated	O
into	O
the	O
walls	O
of	O
long	O
-	O
fibril	O
expanded	O
polytetrafluoroethylene	B-Simple_chemical
(	O
ePTFE	B-Simple_chemical
)	O
vascular	B-Multi-tissue_structure
grafts	I-Multi-tissue_structure
.	O

The	O
grafts	B-Multi-tissue_structure
were	O
implanted	O
in	O
the	O
abdominal	B-Multi-tissue_structure
aortic	I-Multi-tissue_structure
position	O
of	O
24	O
dogs	B-Organism
autologously	O
.	O

Marrow	B-Cell
cells	I-Cell
survived	O
and	O
continued	O
exogenous	O
hemopoiesis	O
for	O
up	O
to	O
six	O
months	O
and	O
were	O
immunohistochemically	O
reactive	O
to	O
basic	B-Gene_or_gene_product
fibroblast	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
bFGF	B-Gene_or_gene_product
)	O
.	O

All	O
the	O
grafts	B-Multi-tissue_structure
older	O
than	O
three	O
weeks	O
had	O
complete	O
endothelialization	O
and	O
maintained	O
their	O
patency	O
.	O

Twenty	O
grafts	B-Multi-tissue_structure
without	O
bone	B-Multi-tissue_structure
marrow	I-Multi-tissue_structure
were	O
implanted	O
as	O
controls	O
.	O

Endothelialization	O
was	O
present	O
at	O
anastomotic	O
sites	O
,	O
but	O
other	O
areas	O
were	O
covered	O
with	O
fresh	O
thrombi	O
.	O

Four	O
out	O
of	O
seven	O
control	O
grafts	B-Multi-tissue_structure
were	O
patent	O
with	O
endothelial	B-Cell
cell	I-Cell
lining	O
at	O
six	O
months	O
,	O
but	O
three	O
were	O
occluded	O
and	O
one	O
of	O
the	O
four	O
grafts	B-Multi-tissue_structure
was	O
still	O
covered	O
with	O
a	O
thrombus	B-Pathological_formation
layer	O
.	O

Bone	B-Multi-tissue_structure
marrow	I-Multi-tissue_structure
with	O
its	O
unique	O
native	O
properties	O
produced	O
autocrine	O
angiogenicity	O
in	O
the	O
graft	O
.	O

Application	O
of	O
clonal	B-Organism
analysis	O
.	O

Differential	O
diagnosis	O
for	O
synchronous	O
primary	O
ovarian	B-Cancer
and	O
endometrial	B-Cancer
cancers	I-Cancer
and	O
metastatic	O
cancer	B-Cancer
.	O

Simultaneous	O
involvement	O
of	O
the	O
endometrium	B-Multi-tissue_structure
and	O
the	O
ovary	B-Organ
by	O
carcinoma	B-Cancer
is	O
a	O
familiar	O
problem	O
in	O
the	O
routine	O
practice	O
of	O
surgical	O
pathology	O
.	O

Such	O
cases	O
may	O
be	O
considered	O
either	O
examples	O
of	O
a	O
single	O
primary	B-Cancer
carcinoma	I-Cancer
with	O
metastasis	O
or	O
as	O
synchronous	B-Cancer
primary	I-Cancer
neoplasms	I-Cancer
.	O

The	O
distinction	O
between	O
these	O
two	O
possibilities	O
is	O
made	O
based	O
on	O
clinicopathologic	O
observations	O
,	O
and	O
therefore	O
may	O
not	O
be	O
definitive	O
.	O

In	O
the	O
present	O
study	O
,	O
the	O
authors	O
used	O
molecular	O
techniques	O
to	O
analyze	O
the	O
clonal	O
composition	O
of	O
five	O
cases	O
of	O
concurrent	O
adenocarcinomas	B-Cancer
of	O
the	O
endometrium	B-Multi-tissue_structure
and	O
ovary	B-Organ
that	O
were	O
clinicopathologically	O
diagnosed	O
as	O
synchronous	B-Cancer
primary	I-Cancer
tumors	I-Cancer
.	O

Patterns	O
of	O
X	B-Cellular_component
-	I-Cellular_component
chromosome	I-Cellular_component
inactivation	O
,	O
mutations	O
in	O
the	O
K	B-Gene_or_gene_product
-	I-Gene_or_gene_product
ras	I-Gene_or_gene_product
gene	O
,	O
mutations	O
or	O
allelic	O
loss	O
of	O
the	O
p53	B-Gene_or_gene_product
gene	O
,	O
or	O
human	B-Organism
papillomavirus	I-Organism
detection	O
were	O
identical	O
in	O
both	O
endometrial	B-Pathological_formation
and	O
ovarian	B-Pathological_formation
lesions	I-Pathological_formation
in	O
three	O
of	O
the	O
cases	O
suggesting	O
that	O
those	O
three	O
cases	O
represented	O
single	O
primary	B-Cancer
tumors	I-Cancer
with	O
metastases	O
.	O

In	O
both	O
of	O
the	O
other	O
two	O
cases	O
,	O
the	O
patterns	O
of	O
X	B-Cellular_component
-	I-Cellular_component
chromosome	I-Cellular_component
inactivation	O
clearly	O
demonstrated	O
the	O
presence	O
of	O
independent	O
primary	B-Cancer
tumors	I-Cancer
.	O

The	O
application	O
of	O
molecular	O
technology	O
may	O
play	O
an	O
important	O
role	O
for	O
the	O
differential	O
diagnosis	O
between	O
synchronous	B-Cancer
primary	I-Cancer
carcinomas	I-Cancer
and	O
a	O
single	O
carcinoma	B-Cancer
with	O
metastasis	O
.	O

Molecular	O
biology	O
of	O
cervical	B-Cancer
cancer	I-Cancer
and	O
its	O
precursors	B-Pathological_formation
.	O

Cervical	B-Cancer
cancer	I-Cancer
develops	O
from	O
well	O
-	O
defined	O
precursor	B-Pathological_formation
lesions	I-Pathological_formation
referred	O
to	O
as	O
either	O
cervical	B-Pathological_formation
intraepithelial	I-Pathological_formation
neoplasia	I-Pathological_formation
or	O
squamous	B-Pathological_formation
intraepithelial	I-Pathological_formation
lesions	I-Pathological_formation
.	O

It	O
is	O
now	O
known	O
that	O
specific	O
types	O
of	O
human	B-Organism
papillomaviruses	I-Organism
(	O
HPV	B-Organism
)	O
are	O
the	O
principal	O
etiologic	O
agents	O
for	O
both	O
cervical	B-Cancer
cancer	I-Cancer
and	O
its	O
precursors	B-Pathological_formation
.	O

The	O
high	O
-	O
oncogenic	O
-	O
risk	O
HPV	B-Organism
types	O
associated	O
with	O
invasive	O
cervical	B-Cancer
cancer	I-Cancer
produce	O
two	O
oncoproteins	O
,	O
designated	O
E6	B-Gene_or_gene_product
and	O
E7	B-Gene_or_gene_product
,	O
which	O
interact	O
with	O
endogenous	O
cell	B-Cell
cycle	O
regulatory	O
proteins	O
,	O
including	O
p53	B-Gene_or_gene_product
and	O
Rb	B-Gene_or_gene_product
.	O

The	O
interaction	O
of	O
virally	O
derived	O
and	O
endogenous	O
cellular	B-Cell
proteins	O
converges	O
in	O
deregulation	O
of	O
cell	B-Cell
cycle	O
progression	O
and	O
appears	O
to	O
be	O
critical	O
for	O
the	O
development	O
of	O
cervical	B-Cancer
cancers	I-Cancer
.	O

However	O
,	O
the	O
development	O
of	O
cervical	B-Cancer
cancer	I-Cancer
is	O
a	O
multistep	O
process	O
that	O
cannot	O
be	O
explained	O
simply	O
by	O
infection	O
with	O
specific	O
types	O
of	O
HPV	B-Organism
.	O

One	O
additional	O
event	O
that	O
appears	O
to	O
play	O
a	O
role	O
in	O
tumor	B-Cancer
progression	O
is	O
integration	O
of	O
HPV	B-Organism
DNA	B-Cellular_component
into	O
the	O
host	O
genome	O
.	O

Integration	O
of	O
HPV	B-Organism
DNA	B-Cellular_component
frequently	O
disrupts	O
the	O
E2	B-Gene_or_gene_product
open	O
reading	O
frames	O
,	O
resulting	O
in	O
overexpression	O
of	O
the	O
E6	B-Gene_or_gene_product
and	O
E7	B-Gene_or_gene_product
oncoproteins	O
and	O
possibly	O
causing	O
genomic	O
instability	O
.	O

Additional	O
cofactors	O
and	O
mutational	O
events	O
may	O
be	O
important	O
in	O
the	O
pathogenesis	O
of	O
invasive	O
cervical	B-Cancer
cancers	I-Cancer
and	O
may	O
include	O
chromosomal	B-Cellular_component
rearrangements	O
,	O
loss	O
of	O
constitutional	O
heterozygosity	O
,	O
and	O
proto	O
-	O
oncogene	O
activation	O
.	O

The	O
effects	O
of	O
radiation	O
on	O
neovascularization	O
in	O
a	O
rat	B-Organism
model	O
.	O

It	O
is	O
thought	O
that	O
radiation	O
treatment	O
inhibits	O
neovascularization	O
of	O
recipient	O
and	O
/	O
or	O
graft	B-Tissue
tissues	I-Tissue
,	O
and	O
this	O
may	O
account	O
in	O
part	O
for	O
abnormalities	O
in	O
wound	B-Pathological_formation
healing	O
associated	O
with	O
radiation	O
therapy	O
.	O

We	O
have	O
examined	O
this	O
hypothesis	O
using	O
a	O
model	O
that	O
measures	O
the	O
neovascularization	O
of	O
an	O
implanted	O
foreign	O
material	O
.	O

Expanded	O
polytetrafluoroethylene	B-Simple_chemical
(	O
PTFE	B-Simple_chemical
)	O
sheets	O
were	O
implanted	O
adjacent	O
to	O
both	O
superficial	O
epigastric	B-Multi-tissue_structure
vascular	I-Multi-tissue_structure
pedicles	I-Multi-tissue_structure
of	O
63	O
rats	B-Organism
distributed	O
into	O
7	O
groups	O
(	O
n	O
=	O
7	O
)	O
that	O
differed	O
with	O
respect	O
to	O
dose	O
and	O
timing	O
of	O
irradiation	O
.	O

Zero	O
to	O
10	O
daily	O
fractions	O
of	O
electron	O
-	O
beam	O
radiation	O
(	O
300	O
cGy	O
each	O
)	O
were	O
delivered	O
to	O
the	O
implant	O
in	O
the	O
right	B-Organism_subdivision
groin	I-Organism_subdivision
,	O
while	O
the	O
implant	O
in	O
the	O
left	B-Organism_subdivision
groin	I-Organism_subdivision
served	O
as	O
a	O
nonirradiated	O
internal	O
control	O
.	O

Unirradiated	O
animals	O
showed	O
equal	O
neovascularization	O
of	O
both	O
implants	O
.	O

Rats	B-Organism
that	O
were	O
irradiated	O
twice	O
(	O
single	O
fractions	O
at	O
0	O
and	O
24	O
hours	O
after	O
implantation	O
)	O
did	O
not	O
show	O
a	O
significant	O
decrease	O
in	O
the	O
neovascularization	O
of	O
the	O
irradiated	O
implant	O
compared	O
with	O
the	O
contralateral	O
control	O
implant	O
.	O

In	O
contrast	O
,	O
the	O
implants	O
that	O
were	O
irradiated	O
three	O
times	O
(	O
single	O
fractions	O
at	O
0	O
,	O
24	O
,	O
and	O
48	O
hours	O
after	O
implantation	O
)	O
demonstrated	O
significantly	O
diminished	O
(	O
greater	O
than	O
25	O
percent	O
,	O
p	O
less	O
than	O
0	O
.	O
05	O
)	O
neovascularization	O
beyond	O
day	O
7	O
,	O
whereas	O
implants	O
irradiated	O
only	O
at	O
48	O
hours	O
after	O
implantation	O
did	O
not	O
.	O

Interestingly	O
,	O
neovascularization	O
of	O
the	O
implants	O
irradiated	O
with	O
10	O
fractions	O
(	O
3000	O
cGy	O
)	O
was	O
not	O
significantly	O
decreased	O
compared	O
with	O
irradiation	O
with	O
three	O
fractions	O
(	O
900	O
cGy	O
)	O
.	O

Irradiation	O
delivered	O
before	O
implantation	O
(	O
900	O
cGy	O
)	O
inhibited	O
neovascularization	O
significantly	O
less	O
than	O
the	O
same	O
dose	O
administered	O
after	O
implantation	O
.	O

The	O
results	O
of	O
this	O
study	O
suggest	O
that	O
a	O
subclinical	O
cumulative	O
dose	O
of	O
900	O
cGy	O
is	O
the	O
threshold	O
for	O
impaired	O
tissue	B-Tissue
revascularization	O
provided	O
that	O
treatment	O
is	O
delivered	O
immediately	O
after	O
implantation	O
over	O
a	O
48	O
-	O
hour	O
interval	O
.	O

Prognostic	O
value	O
of	O
angiogenesis	O
in	O
operable	O
non	B-Cancer
-	I-Cancer
small	I-Cancer
cell	I-Cancer
lung	I-Cancer
cancer	I-Cancer
.	O

Tumour	B-Cancer
angiogenesis	O
is	O
an	O
important	O
factor	O
for	O
tumour	B-Cancer
growth	O
and	O
metastasis	O
.	O

Although	O
some	O
recent	O
reports	O
suggest	O
that	O
microvessel	B-Tissue
counts	O
in	O
non	B-Cancer
-	I-Cancer
small	I-Cancer
cell	I-Cancer
lung	I-Cancer
cancer	I-Cancer
are	O
related	O
to	O
a	O
poor	O
disease	O
outcome	O
,	O
the	O
results	O
were	O
not	O
conclusive	O
and	O
were	O
not	O
compared	O
with	O
other	O
molecular	O
prognostic	O
markers	O
.	O

In	O
the	O
present	O
study	O
,	O
the	O
vascular	B-Multi-tissue_structure
grade	O
was	O
assessed	O
in	O
107	O
(	O
T1	O
,	O
2	O
-	O
N0	O
,	O
1	O
)	O
operable	O
non	B-Cancer
-	I-Cancer
small	I-Cancer
cell	I-Cancer
lung	I-Cancer
carcinomas	I-Cancer
,	O
using	O
the	O
JC70	B-Gene_or_gene_product
monoclonal	I-Gene_or_gene_product
antibody	I-Gene_or_gene_product
to	I-Gene_or_gene_product
CD31	I-Gene_or_gene_product
.	O

Three	O
vascular	B-Multi-tissue_structure
grades	O
were	O
defined	O
with	O
appraisal	O
by	O
eye	B-Organ
and	O
by	O
Chalkley	O
counting	O
:	O
high	O
(	O
Chalkley	O
score	O
7	O
-	O
12	O
)	O
,	O
medium	O
(	O
5	O
-	O
6	O
)	O
,	O
and	O
low	O
(	O
2	O
-	O
4	O
)	O
.	O

There	O
was	O
a	O
significant	O
correlation	O
between	O
eye	B-Organ
appraisal	O
and	O
Chalkley	O
counting	O
(	O
P	O
less	O
than	O
0	O
.	O
0001	O
)	O
.	O

Vascular	B-Multi-tissue_structure
grade	O
was	O
not	O
related	O
to	O
histology	O
,	O
grade	O
,	O
proliferation	O
index	O
(	O
Ki67	B-Gene_or_gene_product
)	O
,	O
or	O
EGFR	B-Gene_or_gene_product
or	O
p53	B-Gene_or_gene_product
expression	O
.	O

Tumours	B-Cancer
from	O
younger	O
patients	B-Organism
had	O
a	O
higher	O
grade	O
of	O
angiogenesis	O
(	O
P	O
=	O
0	O
.	O
05	O
)	O
.	O

Apart	O
from	O
the	O
vascular	B-Multi-tissue_structure
grade	O
,	O
none	O
of	O
the	O
other	O
factors	O
examined	O
was	O
statistically	O
related	O
to	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastasis	O
(	O
P	O
less	O
than	O
0	O
.	O
0001	O
)	O
.	O

A	O
univariate	O
analysis	O
of	O
survival	O
showed	O
that	O
vascular	B-Multi-tissue_structure
grade	O
was	O
the	O
most	O
significant	O
prognostic	O
factor	O
(	O
P	O
=	O
0	O
.	O
0004	O
)	O
,	O
followed	O
by	O
N	O
-	O
stage	O
(	O
P	O
=	O
0	O
.	O
001	O
)	O
.	O

In	O
a	O
multivariate	O
analysis	O
,	O
N	O
-	O
stage	O
and	O
vascular	B-Multi-tissue_structure
grade	O
were	O
not	O
found	O
to	O
be	O
independent	O
prognostic	O
factors	O
,	O
since	O
they	O
were	O
strongly	O
related	O
to	O
each	O
other	O
.	O

Excluding	O
N	O
-	O
stage	O
,	O
vascular	B-Multi-tissue_structure
grade	O
was	O
the	O
only	O
independent	O
prognostic	O
factor	O
(	O
P	O
=	O
0	O
.	O
007	O
)	O
.	O

Kaplan	O
-	O
Meier	O
survival	O
curves	O
showed	O
a	O
statistically	O
significant	O
worse	O
prognosis	O
for	O
patients	B-Organism
with	O
high	O
vascular	B-Multi-tissue_structure
grade	O
,	O
but	O
no	O
difference	O
was	O
observed	O
between	O
low	O
and	O
medium	O
vascular	B-Multi-tissue_structure
grade	O
.	O

These	O
data	O
suggest	O
that	O
angiogenesis	O
in	O
operable	O
non	B-Cancer
-	I-Cancer
small	I-Cancer
cell	I-Cancer
lung	I-Cancer
cancer	I-Cancer
is	O
a	O
major	O
prognostic	O
factor	O
for	O
survival	O
and	O
,	O
among	O
the	O
parameters	O
tested	O
,	O
is	O
the	O
only	O
factor	O
related	O
to	O
cancer	B-Cell
cell	I-Cell
migration	O
to	O
lymph	B-Multi-tissue_structure
nodes	I-Multi-tissue_structure
.	O

The	O
integration	O
of	O
vascular	B-Multi-tissue_structure
grading	O
in	O
clinical	O
trials	O
on	O
adjuvant	O
chemotherapy	O
and	O
/	O
or	O
radiotherapy	O
could	O
substantially	O
contribute	O
in	O
defining	O
groups	O
of	O
operable	O
patients	B-Organism
who	O
might	O
benefit	O
from	O
cytotoxic	O
treatment	O
.	O

Vitamin	B-Simple_chemical
E	I-Simple_chemical
inhibits	O
experimental	O
carcinogenesis	O
and	O
tumour	B-Cancer
angiogenesis	O
.	O

In	O
an	O
experiment	O
in	O
which	O
vitamin	B-Simple_chemical
E	I-Simple_chemical
inhibited	O
carcinogenesis	O
,	O
it	O
was	O
found	O
that	O
tumour	B-Cancer
angiogenesis	O
and	O
tumour	B-Gene_or_gene_product
growth	I-Gene_or_gene_product
-	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
alpha	I-Gene_or_gene_product
(	O
TGF	B-Gene_or_gene_product
alpha	I-Gene_or_gene_product
)	O
expression	O
were	O
also	O
inhibited	O
.	O

Forty	O
male	O
golden	B-Organism
hamsters	I-Organism
were	O
divided	O
into	O
four	O
equal	O
groups	O
.	O

Group	B-Organism
1	I-Organism
animals	I-Organism
had	O
the	O
left	B-Organism_subdivision
buccal	I-Organism_subdivision
pouches	I-Organism_subdivision
painted	O
three	O
times	O
weekly	O
with	O
7	B-Simple_chemical
,	I-Simple_chemical
12	I-Simple_chemical
-	I-Simple_chemical
dimethylbenz	I-Simple_chemical
(	I-Simple_chemical
a	I-Simple_chemical
)	I-Simple_chemical
anthracene	I-Simple_chemical
(	O
DMBA	B-Simple_chemical
)	O
for	O
14	O
weeks	O
.	O

Group	B-Organism
2	I-Organism
animals	I-Organism
had	O
the	O
same	O
procedure	O
of	O
DMBA	B-Simple_chemical
applications	O
but	O
also	O
received	O
alpha	B-Simple_chemical
tocopherol	I-Simple_chemical
.	O

Groups	B-Organism
3	I-Organism
and	O
4	B-Organism
were	O
vitamin	B-Simple_chemical
E	I-Simple_chemical
and	O
untreated	O
controls	O
.	O

Angiogenesis	O
was	O
studied	O
with	O
factor	B-Gene_or_gene_product
8	I-Gene_or_gene_product
-	I-Gene_or_gene_product
related	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
(	O
F8	B-Gene_or_gene_product
-	I-Gene_or_gene_product
RA	I-Gene_or_gene_product
)	O
which	O
identifies	O
endothelial	B-Cell
cells	I-Cell
.	O

TGF	B-Gene_or_gene_product
alpha	I-Gene_or_gene_product
was	O
studied	O
with	O
the	O
appropriate	O
antibody	O
.	O

Staining	O
was	O
effected	O
by	O
the	O
standard	O
avidin	B-Gene_or_gene_product
-	O
biotin	B-Simple_chemical
horseradish	B-Organism
peroxidase	B-Gene_or_gene_product
system	O
.	O

Mean	O
tumour	B-Cancer
volume	O
was	O
significantly	O
lower	O
in	O
the	O
DMBA	B-Simple_chemical
-	O
vitamin	B-Simple_chemical
E	I-Simple_chemical
group	O
compared	O
to	O
the	O
tumour	B-Cancer
control	O
group	O
.	O

Angiogenesis	O
was	O
significantly	O
inhibited	O
in	O
the	O
DMBA	B-Simple_chemical
-	O
vitamin	B-Simple_chemical
E	I-Simple_chemical
group	O
and	O
TGF	B-Gene_or_gene_product
alpha	I-Gene_or_gene_product
expression	O
was	O
also	O
inhibited	O
.	O

It	O
is	O
suggested	O
that	O
inhibition	O
of	O
tumour	B-Cancer
angiogenesis	O
by	O
vitamin	B-Simple_chemical
E	I-Simple_chemical
may	O
be	O
an	O
additional	O
mechanism	O
for	O
the	O
anticancer	B-Cancer
action	O
of	O
vitamin	B-Simple_chemical
E	I-Simple_chemical
.	O

Curcumin	B-Simple_chemical
induces	O
apoptosis	O
in	O
immortalized	O
NIH	B-Cell
3T3	I-Cell
and	O
malignant	B-Cell
cancer	I-Cell
cell	I-Cell
lines	I-Cell
.	O

Curcumin	B-Simple_chemical
,	O
which	O
is	O
a	O
widely	O
used	O
dietary	O
pigment	O
and	O
spice	O
,	O
has	O
been	O
demonstrated	O
to	O
be	O
an	O
effective	O
inhibitor	O
of	O
tumor	B-Cancer
promotion	O
in	O
mouse	B-Organism
skin	B-Organ
carcinogenesis	O
.	O

We	O
report	O
that	O
curcumin	B-Simple_chemical
induces	O
cell	B-Cell
shrinkage	O
,	O
chromatin	B-Cellular_component
condensation	O
,	O
and	O
DNA	B-Cellular_component
fragmentation	O
,	O
characteristics	O
of	O
apoptosis	O
,	O
in	O
immortalized	O
mouse	B-Organism
embryo	B-Developing_anatomical_structure
fibroblast	B-Cell
NIH	I-Cell
3T3	I-Cell
erb	B-Gene_or_gene_product
B2	I-Gene_or_gene_product
oncogene	O
-	O
transformed	O
NIH	B-Cell
3T3	I-Cell
,	O
mouse	B-Organism
sarcoma	B-Cell
S180	I-Cell
,	O
human	B-Organism
colon	B-Cell
cancer	I-Cell
cell	I-Cell
HT	I-Cell
-	I-Cell
29	I-Cell
,	O
human	B-Organism
kidney	B-Cell
cancer	I-Cell
cell	I-Cell
293	I-Cell
,	O
and	O
human	B-Organism
hepatocellular	B-Cell
carcinoma	I-Cell
Hep	I-Cell
G2	I-Cell
cells	I-Cell
,	O
but	O
not	O
in	O
primary	B-Cell
culture	I-Cell
of	O
mouse	B-Organism
embryonic	B-Cell
fibroblast	I-Cell
C3H	I-Cell
10T1	I-Cell
/	I-Cell
2	I-Cell
,	O
rat	B-Organism
embryonic	B-Cell
fibroblast	I-Cell
,	O
and	O
human	B-Organism
foreskin	B-Cell
fibroblast	I-Cell
cells	I-Cell
in	O
a	O
concentration	O
-	O
and	O
time	O
-	O
dependent	O
manner	O
.	O

Many	O
cellular	B-Cell
and	O
biochemical	O
effects	O
of	O
curcumin	B-Simple_chemical
in	O
mouse	B-Organism
fibroblast	B-Cell
cells	I-Cell
have	O
been	O
reported	O
,	O
such	O
as	O
inhibition	O
of	O
protein	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
C	I-Gene_or_gene_product
(	O
PKC	B-Gene_or_gene_product
)	O
activity	O
induced	O
by	O
phorbol	B-Simple_chemical
12	I-Simple_chemical
-	I-Simple_chemical
myristate	I-Simple_chemical
13	I-Simple_chemical
-	I-Simple_chemical
acetate	I-Simple_chemical
treatment	O
,	O
inhibition	O
of	O
tyrosine	B-Amino_acid
protein	O
kinase	O
activity	O
,	O
and	O
inhibition	O
of	O
arachidonic	B-Simple_chemical
acid	I-Simple_chemical
(	O
AA	B-Simple_chemical
)	O
metabolism	O
.	O

Treatment	O
of	O
NIH	B-Cell
3T3	I-Cell
cells	I-Cell
with	O
the	O
PKC	B-Gene_or_gene_product
inhibitor	O
staurosporine	B-Simple_chemical
,	O
the	O
tyrosine	O
kinase	O
inhibitor	O
herbimycin	B-Simple_chemical
A	I-Simple_chemical
,	O
and	O
the	O
AA	B-Simple_chemical
metabolism	O
inhibitor	O
quinacrine	B-Simple_chemical
induces	O
apoptotic	O
cell	B-Cell
death	O
.	O

These	O
results	O
suggest	O
that	O
,	O
in	O
some	O
immortalized	O
and	O
transformed	O
cells	B-Cell
,	O
blocking	O
the	O
cellular	B-Cell
signal	O
transduction	O
might	O
trigger	O
the	O
induction	O
of	O
apoptosis	O
.	O

Fas	B-Gene_or_gene_product
-	O
signaling	O
and	O
effects	O
on	O
receptor	B-Gene_or_gene_product
tyrosine	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
signal	O
transduction	O
in	O
human	B-Organism
breast	B-Cell
epithelial	I-Cell
cells	I-Cell
.	O

Fas	B-Gene_or_gene_product
-	O
mediated	O
cell	B-Cell
death	O
was	O
examined	O
in	O
MCF	B-Cell
-	I-Cell
10AT	I-Cell
preneoplastic	I-Cell
human	I-Cell
breast	I-Cell
epithelial	I-Cell
cells	I-Cell
.	O

Treatment	O
with	O
anti	O
-	O
Fas	B-Gene_or_gene_product
for	O
48	O
h	O
induced	O
apoptosis	O
with	O
cells	B-Cell
exhibiting	O
typical	O
apoptotic	O
features	O
including	O
loss	O
of	O
cell	B-Cell
contact	O
,	O
condensation	O
of	O
chromatin	B-Cellular_component
,	O
and	O
increased	O
staining	O
of	O
the	O
nuclear	B-Cellular_component
membrane	I-Cellular_component
.	O

DNA	B-Cellular_component
fragmentation	O
occurred	O
in	O
response	O
to	O
anti	O
-	O
Fas	B-Gene_or_gene_product
treatment	O
.	O

Anti	O
-	O
Fas	B-Gene_or_gene_product
treatment	O
resulted	O
in	O
decreased	O
p53	B-Gene_or_gene_product
protein	O
levels	O
,	O
while	O
bcl	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
and	O
bax	B-Gene_or_gene_product
protein	O
levels	O
remained	O
unaffected	O
.	O

Cells	B-Cell
treated	O
with	O
anti	O
-	O
Fas	B-Gene_or_gene_product
also	O
exhibited	O
increased	O
tyrosine	B-Amino_acid
phosphorylation	O
of	O
the	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
met	I-Gene_or_gene_product
growth	O
factor	O
receptor	O
tyrosine	B-Amino_acid
kinase	O
.	O

Immunoprecipitation	O
experiments	O
demonstrated	O
that	O
Fas	B-Gene_or_gene_product
associated	O
with	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
erbB2	I-Gene_or_gene_product
and	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
met	I-Gene_or_gene_product
in	O
untreated	O
cells	B-Cell
.	O

Treatment	O
with	O
anti	O
-	O
Fas	B-Gene_or_gene_product
,	O
however	O
,	O
significantly	O
decreased	O
Fas	B-Gene_or_gene_product
-	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
erbB2	I-Gene_or_gene_product
and	O
Fas	B-Gene_or_gene_product
-	O
c	B-Gene_or_gene_product
-	I-Gene_or_gene_product
met	I-Gene_or_gene_product
association	O
.	O

Anti	O
-	O
Fas	B-Gene_or_gene_product
treatment	O
of	O
these	O
cells	B-Cell
caused	O
a	O
significant	O
decrease	O
in	O
p120	B-Gene_or_gene_product
-	O
GAP	B-Gene_or_gene_product
levels	O
,	O
ERK	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
levels	O
,	O
and	O
phosphorylation	O
,	O
as	O
well	O
as	O
Grb2	B-Gene_or_gene_product
-	O
Sosl	B-Gene_or_gene_product
and	O
MEK	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
ERK	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
association	O
.	O

These	O
results	O
show	O
that	O
Fas	B-Gene_or_gene_product
-	O
signaling	O
exerted	O
a	O
suppressive	O
effect	O
on	O
p53	B-Gene_or_gene_product
levels	O
and	O
on	O
downstream	O
effectors	O
of	O
receptor	B-Gene_or_gene_product
tyrosine	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
signal	O
transduction	O
,	O
thereby	O
ensuring	O
cell	B-Cell
death	O
.	O

Prevention	O
of	O
hepatic	B-Cancer
tumor	I-Cancer
metastases	O
in	O
rats	B-Organism
with	O
herpes	B-Organism
viral	I-Organism
vaccines	O
and	O
gamma	B-Gene_or_gene_product
-	I-Gene_or_gene_product
interferon	I-Gene_or_gene_product
.	O

Previous	O
studies	O
showed	O
that	O
gammaIFN	B-Gene_or_gene_product
decreases	O
metastatic	O
hepatic	B-Cancer
tumor	I-Cancer
growth	O
by	O
stimulating	O
Kupffer	B-Cell
cells	I-Cell
(	O
KC	B-Cell
)	O
.	O

The	O
present	O
studies	O
examine	O
whether	O
lymphocyte	B-Cell
stimulation	O
via	O
cells	B-Cell
engineered	O
to	O
secrete	O
GM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
or	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
decreases	O
hepatic	B-Cancer
tumor	I-Cancer
growth	O
,	O
and	O
whether	O
stimulation	O
of	O
both	O
macrophages	B-Cell
and	O
lymphocytes	B-Cell
is	O
more	O
effective	O
than	O
either	O
individually	O
.	O

Rats	B-Organism
were	O
immunized	O
with	O
irradiated	O
hepatoma	B-Cell
cells	I-Cell
transduced	O
by	O
herpes	B-Organism
viral	I-Organism
amplicon	O
vectors	O
containing	O
the	O
genes	O
for	O
GM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
,	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
or	O
LacZ	B-Gene_or_gene_product
.	O

On	O
day	O
18	O
,	O
half	O
of	O
each	O
group	B-Organism
was	O
treated	O
with	O
5	O
x	O
10	O
(	O
4	O
)	O
U	O
gammaIFN	B-Gene_or_gene_product
,	O
or	O
saline	B-Simple_chemical
intraperitoneally	B-Immaterial_anatomical_entity
for	O
3	O
d	O
.	O

On	O
day	O
21	O
,	O
all	O
rats	B-Organism
received	O
5	O
x	O
10	O
(	O
5	O
)	O
hepatoma	B-Cell
cells	I-Cell
intrasplenically	B-Immaterial_anatomical_entity
.	O

On	O
day	O
41	O
,	O
rats	B-Organism
were	O
killed	O
and	O
tumor	B-Cancer
nodules	I-Cancer
were	O
counted	O
.	O

Separate	O
rats	B-Organism
underwent	O
splenocyte	B-Cell
and	O
KC	B-Cell
harvest	O
for	O
assessment	O
of	O
lymphocyte	B-Cell
-	O
and	O
macrophage	B-Cell
-	O
mediated	O
tumor	B-Cell
cell	I-Cell
kill	O
in	O
vitro	O
.	O

GM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
or	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
vaccines	O
or	O
gammaIFN	B-Gene_or_gene_product
decreased	O
tumor	B-Cancer
nodules	I-Cancer
significantly	O
(	O
GM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
13	O
+	O
/	O
-	O
4	O
,	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
14	O
+	O
/	O
-	O
6	O
vs	O
.	O
control	O
75	O
+	O
/	O
-	O
24	O
,	O
P	O
<	O
0	O
.	O
001	O
)	O
.	O

Combination	O
therapy	O
was	O
more	O
effective	O
,	O
and	O
completely	O
eliminated	O
tumor	B-Cancer
in	O
4	O
of	O
12	O
IFN	B-Gene_or_gene_product
-	I-Gene_or_gene_product
GM	I-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
and	O
8	O
of	O
11	O
IFN	B-Gene_or_gene_product
-	I-Gene_or_gene_product
IL	I-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
animals	O
.	O

Additional	O
rats	B-Organism
underwent	O
partial	O
hepatectomy	O
,	O
an	O
immunosuppressive	O
procedure	O
known	O
to	O
accelerate	O
the	O
growth	O
of	O
hepatic	B-Cancer
tumor	I-Cancer
,	O
following	O
tumor	B-Cancer
challenge	O
.	O

Therapy	O
was	O
equally	O
effective	O
in	O
this	O
immunosuppressive	O
setting	O
.	O

Vaccination	O
is	O
associated	O
with	O
enhancement	O
of	O
splenocyte	B-Cell
-	O
mediated	O
tumoricidal	O
activity	O
,	O
whereas	O
the	O
effect	O
of	O
gammaIFN	B-Gene_or_gene_product
is	O
mediated	O
by	O
KC	B-Cell
.	O

GM	B-Gene_or_gene_product
-	I-Gene_or_gene_product
CSF	I-Gene_or_gene_product
and	O
IL	B-Gene_or_gene_product
-	I-Gene_or_gene_product
2	I-Gene_or_gene_product
vaccine	O
therapy	O
and	O
pretreatment	O
with	O
gammaIFN	B-Gene_or_gene_product
represent	O
effective	O
strategies	O
in	O
reducing	O
hepatic	B-Cancer
tumor	I-Cancer
.	O

Combination	O
therapy	O
targets	O
both	O
lymphocytes	B-Cell
and	O
macrophages	B-Cell
,	O
and	O
is	O
more	O
effective	O
in	O
reducing	O
tumor	B-Cancer
than	O
either	O
therapy	O
alone	O
.	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	I-Gene_or_gene_product
induces	O
tumor	B-Cancer
hexokinase	B-Gene_or_gene_product
RNA	O
expression	O
in	O
cancer	B-Cell
cells	I-Cell
.	O

Increased	O
glycolysis	O
is	O
a	O
characteristic	O
of	O
cancer	B-Cell
cells	I-Cell
.	O

Though	O
less	O
efficient	O
in	O
energy	O
production	O
,	O
it	O
ensures	O
continuous	O
supply	O
of	O
energy	O
and	O
phosphometabolites	O
for	O
biosynthesis	O
enabling	O
metastatic	B-Cell
and	O
less	O
vascularized	B-Cell
cancer	I-Cell
cells	I-Cell
to	O
proliferate	O
even	O
under	O
hypoxic	O
conditions	O
.	O

Since	O
hexokinase	B-Gene_or_gene_product
is	O
the	O
first	O
rate	O
limiting	O
enzyme	O
in	O
the	O
glycolytic	O
pathway	O
,	O
elevated	O
levels	O
of	O
Type	B-Gene_or_gene_product
II	I-Gene_or_gene_product
like	I-Gene_or_gene_product
hexokinase	I-Gene_or_gene_product
in	O
tumors	B-Cancer
are	O
of	O
great	O
significance	O
in	O
this	O
context	O
.	O

Under	O
normal	O
conditions	O
insulin	B-Gene_or_gene_product
regulates	O
expression	O
of	O
hexokinase	B-Gene_or_gene_product
Type	I-Gene_or_gene_product
II	I-Gene_or_gene_product
isoenzyme	O
,	O
which	O
is	O
predominantly	O
expressed	O
in	O
muscle	B-Organ
.	O

On	O
the	O
other	O
hand	O
cancer	B-Cell
cells	I-Cell
overexpress	O
insulin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
like	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factors	I-Gene_or_gene_product
and	O
their	O
receptors	O
which	O
mimic	O
many	O
activities	O
of	O
insulin	B-Gene_or_gene_product
.	O

This	O
prompted	O
us	O
to	O
examine	O
a	O
hypothesis	O
that	O
insulin	B-Gene_or_gene_product
-	I-Gene_or_gene_product
like	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factors	I-Gene_or_gene_product
may	O
be	O
responsible	O
for	O
overexpression	O
of	O
tumor	B-Cancer
hexokinase	B-Gene_or_gene_product
.	O

Our	O
experiments	O
demonstrate	O
that	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	I-Gene_or_gene_product
indeed	O
induces	O
hexokinase	B-Gene_or_gene_product
gene	O
expression	O
in	O
a	O
concentration	O
and	O
time	O
dependent	O
manner	O
in	O
two	O
cancer	B-Cell
cell	I-Cell
lines	I-Cell
we	O
studied	O
.	O

Suppression	O
of	O
tumor	B-Cancer
growth	O
by	O
the	O
3	O
'	O
untranslated	O
region	O
of	O
mel	B-Gene_or_gene_product
-	I-Gene_or_gene_product
18	I-Gene_or_gene_product
in	O
3Y1	B-Cell
cells	I-Cell
transformed	O
by	O
the	O
E6	B-Gene_or_gene_product
and	O
E7	B-Gene_or_gene_product
genes	O
of	O
human	B-Organism
papillomavirus	I-Organism
type	I-Organism
18	I-Organism
.	O

By	O
introducing	O
a	O
cDNA	O
library	O
derived	O
from	O
rat	B-Organism
embryonic	B-Cell
fibroblast	I-Cell
cells	I-Cell
,	O
we	O
isolated	O
several	O
morphologically	O
flat	O
revertants	O
of	O
rat	B-Organism
3Y1	B-Cell
cells	I-Cell
transformed	O
by	O
the	O
E6	B-Gene_or_gene_product
and	O
E7	B-Gene_or_gene_product
genes	O
of	O
human	B-Organism
papillomavirus	I-Organism
type	I-Organism
18	I-Organism
(	O
HPV18	B-Organism
)	O
.	O

From	O
one	O
of	O
the	O
revertants	O
,	O
we	O
recovered	O
a	O
0	O
.	O
2	O
-	O
kb	O
cDNA	O
,	O
N56	B-Gene_or_gene_product
,	O
that	O
suppresses	O
the	O
tumor	B-Cancer
growth	O
of	O
the	O
transformed	B-Cell
3Y1	I-Cell
cells	I-Cell
irrespective	O
of	O
the	O
expression	O
of	O
the	O
E6	B-Gene_or_gene_product
and	O
E7	B-Gene_or_gene_product
genes	O
.	O

The	O
nucleotide	O
sequence	O
of	O
the	O
cDNA	O
was	O
shown	O
to	O
be	O
identical	O
to	O
that	O
of	O
the	O
3	O
'	O
untranslated	O
region	O
of	O
a	O
putative	O
mammalian	B-Organism
polycomb	O
group	O
gene	O
,	O
mel	B-Gene_or_gene_product
-	I-Gene_or_gene_product
18	I-Gene_or_gene_product
.	O

Calretinin	B-Gene_or_gene_product
.	O

A	O
selective	O
marker	O
of	O
normal	O
and	O
neoplastic	B-Cell
mesothelial	I-Cell
cells	I-Cell
in	O
serous	B-Organism_substance
effusions	I-Organism_substance
.	O

OBJECTIVE	O
:	O
To	O
document	O
that	O
a	O
polyclonal	O
antiserum	B-Organism_substance
to	O
calretinin	B-Gene_or_gene_product
,	O
a	O
29	O
-	O
kd	O
calcium	B-Simple_chemical
-	O
binding	O
protein	O
,	O
consistently	O
decorates	O
normal	O
and	O
tumor	B-Cell
mesothelial	I-Cell
cells	I-Cell
in	O
cytologic	B-Tissue
preparations	I-Tissue
.	O

STUDY	O
DESIGN	O
:	O
Thirty	O
-	O
three	O
archival	O
cytologic	B-Tissue
specimens	I-Tissue
from	O
eight	O
patients	B-Organism
with	O
histologically	O
confirmed	O
malignant	B-Cancer
mesothelioma	I-Cancer
and	O
13	O
from	O
patients	B-Organism
with	O
metastatic	O
serous	B-Organism_substance
effusions	I-Organism_substance
were	O
destained	O
and	O
then	O
immunostained	O
with	O
anticalretinin	O
antiserum	B-Organism_substance
.	O

For	O
investigation	O
of	O
cell	B-Cell
suspensions	I-Cell
,	O
four	O
pleural	B-Organism_substance
fluids	I-Organism_substance
were	O
incubated	O
with	O
anticalretinin	O
antiserum	B-Organism_substance
.	O

After	O
cytocentrifugation	O
the	O
specimens	B-Multi-tissue_structure
were	O
stained	O
in	O
accordance	O
with	O
the	O
alkaline	B-Gene_or_gene_product
phosphatase	I-Gene_or_gene_product
anti	O
-	O
alkaline	B-Gene_or_gene_product
phosphatase	I-Gene_or_gene_product
(	O
APAAP	O
)	O
method	O
.	O

For	O
electron	O
microscopic	O
examination	O
the	O
cell	B-Cell
suspensions	I-Cell
were	O
then	O
incubated	O
with	O
gold	B-Simple_chemical
-	O
labeled	O
antirabbit	O
antibody	O
.	O

RESULTS	O
:	O
The	O
diagnostic	O
sensitivity	O
of	O
this	O
new	O
immunocytochemical	O
approach	O
reached	O
100	O
%	O
for	O
the	O
eight	O
malignant	B-Cancer
mesotheliomas	I-Cancer
investigated	O
.	O

Only	O
3	O
of	O
the	O
13	O
adenocarcinomas	B-Cancer
metastatic	O
to	O
the	O
serous	B-Organism_substance
membranes	I-Organism_substance
included	O
in	O
this	O
study	O
were	O
weakly	O
reactive	O
,	O
accounting	O
for	O
81	O
%	O
specificity	O
.	O

Binding	O
of	O
anticalretinin	O
antiserum	B-Organism_substance
to	O
living	O
mesothelial	B-Cell
cells	I-Cell
was	O
consistently	O
documented	O
in	O
all	O
four	O
cases	O
investigated	O
.	O

CONCLUSION	O
:	O
Calretinin	B-Gene_or_gene_product
is	O
a	O
very	O
useful	O
marker	O
for	O
positive	O
identification	O
of	O
normal	O
and	O
tumor	B-Cell
mesothelial	I-Cell
cells	I-Cell
in	O
serous	B-Organism_substance
effusions	I-Organism_substance
.	O

Prognostic	O
value	O
of	O
HLA	B-Gene_or_gene_product
-	I-Gene_or_gene_product
DR	I-Gene_or_gene_product
expression	O
and	O
dendritic	B-Cell
cell	I-Cell
infiltration	O
in	O
gastric	B-Cancer
cancer	I-Cancer
.	O

We	O
attempted	O
to	O
correlate	O
the	O
expression	O
of	O
human	B-Gene_or_gene_product
leukocyte	I-Gene_or_gene_product
antigen	I-Gene_or_gene_product
DR	I-Gene_or_gene_product
(	O
HLA	B-Gene_or_gene_product
-	I-Gene_or_gene_product
DR	I-Gene_or_gene_product
)	O
and	O
tumor	B-Cancer
infiltration	O
by	O
S	B-Gene_or_gene_product
-	I-Gene_or_gene_product
100	I-Gene_or_gene_product
-	O
protein	O
-	O
positive	O
dendritic	O
cells	O
with	O
clinicopathologic	O
features	O
in	O
165	O
patients	B-Organism
with	O
gastric	B-Cancer
cancer	I-Cancer
.	O

The	O
expression	O
of	O
HLA	B-Gene_or_gene_product
-	I-Gene_or_gene_product
DR	I-Gene_or_gene_product
was	O
correlated	O
with	O
the	O
histologic	O
type	O
.	O

Infiltration	O
of	O
dendritic	B-Cell
cells	I-Cell
correlated	O
negatively	O
with	O
distant	O
lymph	B-Multi-tissue_structure
node	I-Multi-tissue_structure
metastases	O
,	O
clinical	O
stage	O
,	O
and	O
peritoneal	B-Multi-tissue_structure
invasion	O
.	O

There	O
was	O
no	O
correlation	O
between	O
the	O
expression	O
of	O
HLA	B-Gene_or_gene_product
-	I-Gene_or_gene_product
DR	I-Gene_or_gene_product
and	O
infiltration	O
by	O
dendritic	B-Cell
cells	I-Cell
.	O

In	O
patients	B-Organism
with	O
resectable	O
gastric	B-Cancer
cancer	I-Cancer
,	O
the	O
grade	O
of	O
infiltrating	O
dendritic	B-Cell
cells	I-Cell
may	O
be	O
a	O
suitable	O
predictor	O
of	O
prognosis	O
.	O

Integrin	B-Gene_or_gene_product
alphavbeta3	I-Gene_or_gene_product
requirement	O
for	O
sustained	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
activity	O
during	O
angiogenesis	O
.	O

Angiogenesis	O
depends	O
on	O
growth	O
factors	O
and	O
vascular	B-Cell
cell	I-Cell
adhesion	O
events	O
.	O

Integrins	B-Gene_or_gene_product
and	O
growth	O
factors	O
are	O
capable	O
of	O
activating	O
the	O
ras	B-Gene_or_gene_product
/	O
MAP	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
pathway	O
in	O
vitro	O
,	O
yet	O
how	O
these	O
signals	O
influence	O
endothelial	B-Cell
cells	I-Cell
during	O
angiogenesis	O
is	O
unknown	O
.	O

Upon	O
initiation	O
of	O
angiogenesis	O
with	O
basic	B-Gene_or_gene_product
fibroblast	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
bFGF	B-Gene_or_gene_product
)	O
on	O
the	O
chick	B-Organism
chorioallantoic	B-Multi-tissue_structure
membrane	I-Multi-tissue_structure
(	O
CAM	B-Multi-tissue_structure
)	O
,	O
endothelial	B-Cell
cell	I-Cell
mitogen	B-Gene_or_gene_product
-	I-Gene_or_gene_product
activated	I-Gene_or_gene_product
protein	I-Gene_or_gene_product
(	I-Gene_or_gene_product
MAP	I-Gene_or_gene_product
)	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
(	O
ERK	B-Gene_or_gene_product
)	O
activity	O
was	O
detected	O
as	O
early	O
as	O
5	O
min	O
yet	O
was	O
sustained	O
for	O
at	O
least	O
20	O
h	O
.	O

The	O
initial	O
wave	O
of	O
ERK	B-Gene_or_gene_product
activity	O
(	O
5	O
-	O
120	O
min	O
)	O
was	O
refractory	O
to	O
integrin	B-Simple_chemical
antagonists	I-Simple_chemical
,	O
whereas	O
the	O
sustained	O
activity	O
(	O
4	O
-	O
20	O
h	O
)	O
depended	O
on	O
integrin	B-Gene_or_gene_product
alphavbeta3	I-Gene_or_gene_product
,	O
but	O
not	O
beta1	B-Gene_or_gene_product
integrins	I-Gene_or_gene_product
.	O

Inhibition	O
of	O
MAP	B-Gene_or_gene_product
kinase	I-Gene_or_gene_product
kinase	I-Gene_or_gene_product
(	O
MEK	B-Gene_or_gene_product
)	O
during	O
this	O
sustained	O
alphavbeta3	B-Gene_or_gene_product
-	O
dependent	O
ERK	B-Gene_or_gene_product
signal	O
blocked	O
the	O
formation	O
of	O
new	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
while	O
not	O
influencing	O
preexisting	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
on	O
the	O
CAM	B-Multi-tissue_structure
.	O

Inhibition	O
of	O
MEK	B-Gene_or_gene_product
also	O
blocked	O
growth	O
factor	O
induced	O
migration	O
but	O
not	O
adhesion	O
of	O
endothelial	B-Cell
cells	I-Cell
in	O
vitro	O
.	O

Therefore	O
,	O
angiogenesis	O
depends	O
on	O
sustained	O
ERK	B-Gene_or_gene_product
activity	O
regulated	O
by	O
the	O
ligation	O
state	O
of	O
both	O
a	O
growth	O
factor	O
receptor	O
and	O
integrin	B-Gene_or_gene_product
alphavbeta3	I-Gene_or_gene_product
.	O

Basic	B-Gene_or_gene_product
fibroblast	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
:	O
a	O
missing	O
link	O
between	O
collagen	B-Gene_or_gene_product
VII	I-Gene_or_gene_product
,	O
increased	O
collagenase	B-Gene_or_gene_product
,	O
and	O
squamous	B-Cancer
cell	I-Cancer
carcinoma	I-Cancer
in	O
recessive	O
dystrophic	O
epidermolysis	O
bullosa	O
.	O

BACKGROUND	O
:	O
Patients	B-Organism
with	O
recessive	O
dystrophic	O
epidermolysis	O
bullosa	O
(	O
RDEB	O
)	O
have	O
deficiencies	O
of	O
collagen	B-Gene_or_gene_product
type	I-Gene_or_gene_product
VII	I-Gene_or_gene_product
and	O
have	O
elevated	O
levels	O
of	O
fibroblast	B-Gene_or_gene_product
collagenase	I-Gene_or_gene_product
,	O
and	O
a	O
greatly	O
increased	O
risk	O
of	O
cutaneous	B-Cancer
squamous	I-Cancer
cell	I-Cancer
carcinoma	I-Cancer
.	O

Patients	B-Organism
with	O
other	O
genetic	O
blistering	O
disorders	O
do	O
not	O
have	O
elevated	O
collagenase	B-Gene_or_gene_product
or	O
an	O
increased	O
risk	O
of	O
squamous	B-Cancer
cell	I-Cancer
carcinoma	I-Cancer
,	O
despite	O
chronic	O
wounding	O
.	O

The	O
connection	O
between	O
collagen	B-Gene_or_gene_product
type	I-Gene_or_gene_product
VII	I-Gene_or_gene_product
deficiency	O
,	O
increased	O
collagenase	B-Gene_or_gene_product
,	O
and	O
squamous	B-Cancer
cell	I-Cancer
carcinoma	I-Cancer
is	O
not	O
understood	O
.	O

MATERIALS	O
AND	O
METHODS	O
:	O
Urine	B-Organism_substance
from	O
81	O
patients	B-Organism
with	O
RDEB	O
(	O
39	O
patients	B-Organism
)	O
,	O
junctional	O
epidermolysis	O
bullosa	O
(	O
JEB	O
;	O
12	O
patients	B-Organism
)	O
,	O
and	O
epidermolysis	O
bullosa	O
simplex	O
(	O
EBS	O
;	O
30	O
patients	B-Organism
)	O
,	O
as	O
well	O
as	O
unaffected	O
family	O
members	O
of	O
RDEB	O
patients	B-Organism
(	O
33	O
patients	B-Organism
)	O
,	O
was	O
tested	O
for	O
the	O
presence	O
of	O
basic	B-Gene_or_gene_product
fibroblast	I-Gene_or_gene_product
growth	I-Gene_or_gene_product
factor	I-Gene_or_gene_product
(	O
bFGF	B-Gene_or_gene_product
)	O
using	O
a	O
sensitive	O
radioimmunoassay	O
.	O

These	O
patients	B-Organism
included	O
many	O
who	O
were	O
enrolled	O
in	O
the	O
Epidermolysis	O
Bullosa	O
Registry	O
and	O
others	O
who	O
were	O
referred	O
by	O
their	O
physicians	O
.	O

RESULTS	O
:	O
Fifty	O
-	O
one	O
percent	O
of	O
patients	B-Organism
with	O
RDEB	O
had	O
elevated	O
levels	O
(	O
greater	O
than	O
5000	O
pg	O
/	O
g	O
)	O
of	O
urinary	B-Organism_substance
bFGF	B-Gene_or_gene_product
.	O

In	O
contrast	O
,	O
none	O
of	O
the	O
patients	B-Organism
with	O
JEB	O
had	O
elevated	O
levels	O
of	O
bFGF	B-Gene_or_gene_product
.	O

Twenty	O
-	O
one	O
percent	O
of	O
clinically	O
unaffected	O
family	O
members	O
had	O
elevated	O
levels	O
of	O
bFGF	B-Gene_or_gene_product
,	O
and	O
13	O
%	O
of	O
patients	B-Organism
with	O
EBS	O
had	O
elevated	O
levels	O
of	O
bFGF	B-Gene_or_gene_product
.	O

The	O
frequency	O
of	O
elevated	O
bFGF	B-Gene_or_gene_product
values	O
among	O
all	O
groups	O
was	O
statistically	O
significant	O
(	O
p	O
=	O
0	O
.	O
002	O
)	O
,	O
and	O
the	O
levels	O
of	O
bFGF	B-Gene_or_gene_product
in	O
RDEB	O
patients	B-Organism
were	O
significantly	O
elevated	O
compared	O
with	O
those	O
of	O
other	O
groups	O
(	O
p	O
less	O
than	O
0	O
.	O
05	O
)	O
.	O

CONCLUSIONS	O
:	O
We	O
have	O
found	O
that	O
patients	B-Organism
with	O
RDEB	O
have	O
elevated	O
levels	O
of	O
bFGF	B-Gene_or_gene_product
,	O
which	O
may	O
contribute	O
to	O
increased	O
fibroblast	B-Gene_or_gene_product
collagenase	I-Gene_or_gene_product
and	O
the	O
development	O
of	O
squamous	B-Cancer
cell	I-Cancer
carcinoma	I-Cancer
.	O

These	O
results	O
suggest	O
a	O
novel	O
treatment	O
for	O
RDEB	O
,	O
namely	O
,	O
angiogenesis	O
inhibitors	O
,	O
which	O
may	O
antagonize	O
the	O
effects	O
of	O
bFGF	B-Gene_or_gene_product
in	O
this	O
disorder	O
.	O

There	O
are	O
currently	O
no	O
other	O
means	O
of	O
treatment	O
for	O
this	O
disorder	O
,	O
which	O
has	O
a	O
high	O
morbidity	O
and	O
mortality	O
rate	O
.	O

The	O
myoadipose	B-Multi-tissue_structure
flap	I-Multi-tissue_structure
:	O
a	O
new	O
composite	O
.	O

A	O
prefabricated	O
composite	B-Multi-tissue_structure
fat	I-Multi-tissue_structure
flap	I-Multi-tissue_structure
consisting	O
of	O
muscle	B-Tissue
woven	O
into	O
an	O
anatomically	O
distinct	O
fat	B-Tissue
pad	I-Tissue
was	O
studied	O
in	O
a	O
rabbit	B-Organism
model	O
.	O

In	O
17	O
rabbits	B-Organism
,	O
a	O
2	O
-	O
cm	O
strip	B-Multi-tissue_structure
of	O
latissimus	B-Organ
dorsi	I-Organ
was	O
woven	O
into	O
the	O
parascapular	B-Tissue
fat	I-Tissue
pad	I-Tissue
on	O
one	O
side	O
,	O
with	O
the	O
contralateral	B-Tissue
fat	I-Tissue
pad	I-Tissue
serving	O
as	O
a	O
control	O
.	O

At	O
3	O
weeks	O
,	O
the	O
endogenous	O
blood	B-Organism_substance
supply	O
of	O
both	O
the	O
control	O
and	O
the	O
experimental	O
fat	B-Tissue
pads	I-Tissue
was	O
isolated	O
and	O
ligated	O
,	O
and	O
the	O
composite	B-Multi-tissue_structure
fat	I-Multi-tissue_structure
/	I-Multi-tissue_structure
muscle	I-Multi-tissue_structure
flap	I-Multi-tissue_structure
was	O
transferred	O
to	O
the	O
chest	B-Organism_subdivision
wall	I-Organism_subdivision
.	O

At	O
6	O
weeks	O
,	O
animals	O
were	O
killed	O
,	O
and	O
flaps	B-Multi-tissue_structure
were	O
analyzed	O
for	O
length	O
,	O
width	O
,	O
and	O
weight	O
;	O
perfused	O
with	O
fluorescein	B-Simple_chemical
or	O
lead	B-Simple_chemical
oxide	I-Simple_chemical
;	O
and	O
examined	O
histologically	O
.	O

Significant	O
differences	O
were	O
found	O
between	O
the	O
control	O
and	O
experimental	O
fat	B-Tissue
pads	I-Tissue
with	O
regard	O
to	O
weight	O
and	O
length	O
.	O

Experimental	O
flaps	B-Multi-tissue_structure
were	O
found	O
to	O
be	O
perfused	O
fully	O
with	O
fluorescein	B-Simple_chemical
and	O
lead	B-Simple_chemical
oxide	I-Simple_chemical
;	O
control	O
fat	B-Tissue
pads	I-Tissue
were	O
found	O
not	O
to	O
be	O
perfused	O
.	O

The	O
lead	B-Simple_chemical
oxide	I-Simple_chemical
group	O
revealed	O
extensive	O
growth	O
of	O
blood	B-Multi-tissue_structure
vessels	I-Multi-tissue_structure
from	O
the	O
latissimus	B-Multi-tissue_structure
graft	I-Multi-tissue_structure
into	O
the	O
experimental	O
fat	B-Tissue
pad	I-Tissue
.	O

No	O
vessels	B-Multi-tissue_structure
were	O
visualized	O
in	O
the	O
controls	O
.	O

Finally	O
,	O
sections	B-Multi-tissue_structure
of	O
the	O
control	O
and	O
experimental	O
flaps	B-Multi-tissue_structure
were	O
analyzed	O
histologically	O
.	O

A	O
preponderance	O
of	O
viable	O
fat	B-Tissue
,	O
with	O
evidence	O
of	O
neovascularization	O
,	O
was	O
found	O
in	O
experimental	O
flaps	B-Multi-tissue_structure
,	O
compared	O
with	O
the	O
necrotic	O
fat	B-Tissue
that	O
characterized	O
the	O
controls	O
.	O

We	O
conclude	O
that	O
prefabrication	O
of	O
a	O
fat	B-Multi-tissue_structure
flap	I-Multi-tissue_structure
is	O
possible	O
and	O
may	O
have	O
extensive	O
application	O
in	O
various	O
areas	O
of	O
plastic	O
surgery	O
.	O

Requirement	O
of	O
estrogen	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
expression	O
and	O
function	O
for	O
[	O
12Val	B-Amino_acid
]	O
K	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Ras	I-Gene_or_gene_product
-	O
mediated	O
NIH3T3	B-Cell
cell	I-Cell
transformation	O
.	O

We	O
investigated	O
the	O
biological	O
significance	O
of	O
estrogen	B-Gene_or_gene_product
receptors	I-Gene_or_gene_product
(	O
ERs	B-Gene_or_gene_product
)	O
in	O
NIH3T3	B-Cell
cell	I-Cell
transformation	O
by	O
the	O
[	O
12Val	B-Amino_acid
]	O
K	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Ras	I-Gene_or_gene_product
mutant	O
.	O

This	O
mutant	O
enhanced	O
the	O
steady	O
-	O
level	O
and	O
transcriptional	O
activity	O
of	O
ER	B-Gene_or_gene_product
.	O

Coexpression	O
of	O
the	O
progesterone	B-Gene_or_gene_product
receptor	I-Gene_or_gene_product
with	O
mutant	O
K	B-Gene_or_gene_product
-	I-Gene_or_gene_product
Ras	I-Gene_or_gene_product
led	O
to	O
suppression	O
of	O
tumorigenicity	O
and	O
inhibition	O
of	O
the	O
activation	O
of	O
ER	B-Gene_or_gene_product
.	O

The	O
antisense	O
oligomers	O
complementary	O
to	O
the	O
ER	B-Gene_or_gene_product
suppressed	O
proliferation	O
and	O
transformed	O
phenotypes	O
of	O
K12V	B-Cell
cells	I-Cell
.	O

These	O
observations	O
support	O
the	O
importance	O
of	O
ER	B-Gene_or_gene_product
in	O
Ras	B-Gene_or_gene_product
-	O
mediated	O
cell	B-Cell
transformation	O
.	O

Gene	O
therapy	O
for	O
gliomas	B-Cancer
:	O
p53	B-Gene_or_gene_product
and	O
E2F	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
proteins	O
and	O
the	O
target	O
of	O
apoptosis	O
.	O

Current	O
therapy	O
for	O
glioma	B-Cancer
is	O
suboptimal	O
.	O

The	O
transfer	O
of	O
apoptosis	O
genes	O
to	O
tumors	B-Cancer
constitutes	O
one	O
of	O
the	O
most	O
promising	O
strategies	O
for	O
cancer	B-Cancer
gene	O
therapy	O
.	O

We	O
have	O
previously	O
shown	O
that	O
massive	O
apoptosis	O
occurs	O
when	O
wild	O
-	O
type	O
p53	B-Gene_or_gene_product
or	O
E2F	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
expression	O
is	O
induced	O
in	O
glioma	B-Cancer
.	O

However	O
,	O
the	O
mechanism	O
of	O
action	O
and	O
the	O
efficiency	O
in	O
inducing	O
apoptosis	O
of	O
these	O
two	O
proteins	O
are	O
not	O
similar	O
.	O

Adenovirus	B-Organism
-	O
mediated	O
p53	B-Gene_or_gene_product
gene	O
transfer	O
is	O
ineffective	O
in	O
causing	O
apoptosis	O
in	O
glioma	B-Cell
cells	I-Cell
that	O
retain	O
wild	O
-	O
type	O
p53	B-Gene_or_gene_product
genotype	O
or	O
overexpress	O
the	O
p21	B-Gene_or_gene_product
protein	O
.	O

The	O
p16	B-Gene_or_gene_product
/	O
Rb	B-Gene_or_gene_product
/	O
E2F	B-Gene_or_gene_product
pathway	O
is	O
the	O
most	O
frequent	O
target	O
of	O
genetic	O
alterations	O
in	O
gliomas	B-Cancer
,	O
and	O
therefore	O
constitutes	O
a	O
suitable	O
target	O
for	O
gene	O
therapy	O
strategies	O
.	O

However	O
,	O
the	O
transfer	O
of	O
either	O
the	O
p16	B-Gene_or_gene_product
or	O
Rb	B-Gene_or_gene_product
gene	O
to	O
glioma	B-Cell
cells	I-Cell
results	O
in	O
cytostatic	O
effect	O
.	O

The	O
E2F	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
protein	O
is	O
able	O
to	O
induce	O
generalized	O
apoptosis	O
in	O
gliomas	B-Cancer
independently	O
of	O
the	O
p53	B-Gene_or_gene_product
,	O
p16	B-Gene_or_gene_product
or	O
Rb	B-Gene_or_gene_product
status	O
.	O

In	O
addition	O
,	O
p21	B-Gene_or_gene_product
-	O
or	O
p16	B-Gene_or_gene_product
-	O
mediated	O
growth	O
arrest	O
did	O
not	O
protect	O
glioma	B-Cell
cells	I-Cell
from	O
E2F	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
mediated	O
apoptosis	O
.	O

The	O
apoptotic	O
molecule	O
bax	B-Gene_or_gene_product
is	O
induced	O
in	O
p53	B-Gene_or_gene_product
-	O
mediated	O
apoptosis	O
,	O
but	O
bax	B-Gene_or_gene_product
is	O
not	O
induced	O
in	O
E2F	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
-	O
mediated	O
apoptosis	O
in	O
glioma	B-Cell
cells	I-Cell
.	O

Careful	O
selection	O
of	O
patients	B-Organism
may	O
be	O
necessary	O
before	O
designing	O
therapeutic	O
strategies	O
using	O
either	O
p53	B-Gene_or_gene_product
or	O
E2F	B-Gene_or_gene_product
-	I-Gene_or_gene_product
1	I-Gene_or_gene_product
as	O
a	O
therapeutic	O
tools	O
for	O
glioma	B-Cancer
patients	B-Organism
.	O

Protective	O
function	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
protein	O
against	O
impaired	O
protein	O
processing	O
in	O
renal	B-Cell
carcinoma	I-Cell
cells	I-Cell
.	O

The	O
absence	O
of	O
functional	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
tumor	B-Cancer
suppressor	O
gene	O
leads	O
to	O
the	O
development	O
of	O
neoplasias	O
characteristic	O
of	O
VHL	B-Gene_or_gene_product
disease	O
,	O
including	O
renal	B-Cancer
cell	I-Cancer
carcinoma	I-Cancer
(	O
RCC	B-Cancer
)	O
.	O

Here	O
,	O
we	O
compared	O
the	O
sensitivity	O
of	O
RCC	B-Cell
cells	I-Cell
lacking	O
VHL	B-Gene_or_gene_product
gene	O
function	O
with	O
that	O
of	O
RCC	B-Cell
cells	I-Cell
expressing	O
the	O
wild	O
-	O
type	O
VHL	B-Gene_or_gene_product
gene	O
(	O
wtVHL	B-Gene_or_gene_product
)	O
after	O
exposure	O
to	O
various	O
stresses	O
.	O

While	O
the	O
response	O
to	O
most	O
treatments	O
was	O
not	O
affected	O
by	O
the	O
VHL	B-Gene_or_gene_product
gene	O
status	O
,	O
glucose	B-Simple_chemical
deprivation	O
was	O
found	O
to	O
be	O
much	O
more	O
cytotoxic	O
for	O
RCC	B-Cell
cells	I-Cell
lacking	O
VHL	B-Gene_or_gene_product
gene	O
function	O
than	O
for	O
wtVHL	B-Gene_or_gene_product
-	O
expressing	O
cells	B-Cell
.	O

The	O
heightened	O
sensitivity	O
of	O
VHL	B-Gene_or_gene_product
-	O
deficient	O
cells	B-Cell
was	O
not	O
attributed	O
to	O
dissimilar	O
energy	O
requirements	O
or	O
to	O
differences	O
in	O
glucose	B-Simple_chemical
uptake	O
,	O
but	O
more	O
likely	O
reflects	O
a	O
lesser	O
ability	O
of	O
VHL	B-Gene_or_gene_product
-	O
deficient	O
cells	B-Cell
to	O
handle	O
abnormally	O
processed	O
proteins	O
arising	O
from	O
impaired	O
glycosylation	O
.	O

In	O
support	O
of	O
this	O
hypothesis	O
,	O
other	O
treatments	O
which	O
act	O
through	O
different	O
mechanisms	O
to	O
interfere	O
with	O
protein	O
processing	O
(	O
i	O
.	O
e	O
.	O
,	O
tunicamycin	B-Simple_chemical
,	O
brefeldin	B-Simple_chemical
A	I-Simple_chemical
,	O
and	O
azetidine	B-Simple_chemical
)	O
were	O
also	O
found	O
to	O
be	O
much	O
more	O
toxic	O
for	O
VHL	B-Gene_or_gene_product
-	O
deficient	O
cells	B-Cell
.	O

Furthermore	O
,	O
ubiquitination	O
of	O
cellular	B-Cell
proteins	O
was	O
elevated	O
in	O
VHL	B-Gene_or_gene_product
-	O
deficient	O
cells	B-Cell
,	O
particularly	O
after	O
glucose	B-Simple_chemical
deprivation	O
,	O
supporting	O
a	O
role	O
for	O
the	O
VHL	B-Gene_or_gene_product
gene	O
in	O
ubiquitin	B-Gene_or_gene_product
-	O
mediated	O
proteolysis	O
.	O

Accordingly	O
,	O
the	O
rate	O
of	O
elimination	O
of	O
abnormal	O
proteins	O
was	O
lower	O
in	O
cells	B-Cell
lacking	O
a	O
functional	O
VHL	B-Gene_or_gene_product
gene	O
than	O
in	O
wtVHL	B-Gene_or_gene_product
-	O
expressing	O
cells	B-Cell
.	O

Thus	O
,	O
pVHL	B-Gene_or_gene_product
appears	O
to	O
participate	O
in	O
the	O
elimination	O
of	O
misprocessed	O
proteins	O
,	O
such	O
as	O
those	O
arising	O
in	O
the	O
cell	B-Cell
due	O
to	O
the	O
unavailability	O
of	O
glucose	B-Simple_chemical
or	O
to	O
other	O
stresses	O
.	O

