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Effects of PLCE1 gene silencing by RNA interference on cell cycling and apoptosis in esophageal carcinoma cells.
[esophageal carcinoma]
Esophageal
squamous
cell
carcinoma
(
ESCC
)
is
one
of
the
most
malignancies
with
a
poor
prognosis
.
The
phospholipase
C
?
gene
(
PLCE
1
)
encodes
a
novel
ras-related
protein
effector
mediating
the
effects
of
R-Ras
on
the
actin
cytoskeleton
and
membrane
protrusion
.
However
,
molecular
mechanisms
pertinent
to
ESCC
are
unclear
.
We
therefore
designed
PLCE
1
-
special
small
interfering
RNA
and
transfected
to
esophageal
squamous
cell
(
EC
)
9706
cells
to
investigate
the
effects
of
PLCE
1
gene
silencing
on
the
cell
cycle
and
apoptosis
of
ESCC
and
indicate
its
important
role
in
the
development
of
ESCC
.
Esophageal
cancer
tissue
specimens
and
normal
esophageal
mucosa
were
obtained
and
assayed
by
immunohistochemical
staining
to
confirm
overexpression
of
PLCE
1
in
neoplasias
.
Fluorescence
microscopy
was
used
to
examine
transfection
efficiency
,
while
the
result
of
PLCE
1
silencing
was
examined
by
reverse
transcription
(
RT-PCR
)
.
Flow
cytometry
and
annexin
V
apoptosis
assays
were
used
to
assess
the
cell
cycle
and
apoptosis
,
respectively
.
Expression
of
cyclin
D
1
and
caspase-
3
was
detected
by
Western
-blotting
.
The
level
of
PLCE
1
protein
in
esophageal
cancer
tissue
was
significantly
higher
than
that
in
normal
tissue
.
After
transfection
,
the
expression
of
PLCE
1
mRNA
in
EC
9706
was
significantly
reduced
,
compared
with
the
control
group
.
Furthermore
,
flow
cytometry
results
suggested
that
the
PLCE
1
gene
silencing
arrested
the
cell
cycle
in
the
G
0
/
G
1
phase
;
apoptosis
was
significantly
higher
than
in
the
negative
control
group
and
mock
group
.
PLCE
1
gene
silencing
by
RNAi
resulted
in
decreased
expression
of
cyclin
D
1
and
increased
expression
of
caspase-
3
.
Our
study
suggests
that
PLCE
1
may
be
an
oncogene
and
play
an
important
role
in
esophageal
carcinogenesis
through
regulating
proteins
which
control
cell
cycling
and
apoptosis
.
Diseases
Validation
Diseases presenting
"esophageal carcinogenesis"
symptom
esophageal adenocarcinoma
esophageal carcinoma
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