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Recruited metastasis suppressor NM23-H2 attenuates expression and activity of peroxisome proliferator-activated receptor δ (PPARδ) in human cholangiocarcinoma.
[cholangiocarcinoma]
Peroxisome
proliferator-activated
receptor
δ
(
PPAR
δ
)
is
a
versatile
regulator
of
distinct
biological
processes
and
overexpression
of
PPAR
δ
in
cancer
may
be
partially
related
to
its
suppression
of
its
own
co
-regulators
.
To
determine
whether
recruited
suppressor
proteins
bind
to
and
regulate
PPAR
δ
expression
,
activity
and
PPAR
δ-dependent
cholangiocarcinoma
proliferation
.
Yeast
two
-hybrid
assays
were
done
using
murine
PPAR
δ
as
bait
.
PPARδ
mRNA
expression
was
determined
by
qPCR
.
Protein
expression
was
measured
by
western
blot
.
Immunohistochemistry
and
fluorescence
microscopy
were
used
to
determine
PPARδ
expression
and
co
-localization
with
NDP
Kinase
alpha
(
NM
23
-
H
2
)
.
Cell
proliferation
assays
were
performed
to
determine
cell
numbers
.
Yeast
two
-hybrid
screening
identified
NM
23
-
H
2
as
a
PPARδ
binding
protein
and
their
interaction
was
confirmed
.
Overexpressed
PPAR
δ
or
treatment
with
the
agonist
GW
501516
resulted
in
increased
cell
proliferation
.
NM
23
-
H
2
siRNA
activated
PPARδ
luciferase
promoter
activity
,
upregulated
PPARδ
RNA
and
protein
expression
and
increased
GW
501516
-
stimulated
CCA
growth
.
Overexpression
of
NM
23
-
H
2
inhibited
PPAR
δ
luciferase
promoter
activity
,
downregulated
PPAR
δ
expression
and
AKT
phosphorylation
and
reduced
GW
501516
-
stimulated
CCA
growth
.
We
report
the
novel
association
of
NM
23
-
H
2
with
PPAR
δ
and
the
negative
regulation
of
PPAR
δ
expression
by
NM
23
-
H
2
binding
to
the
C-
terminal
region
of
PPAR
δ
.
These
findings
provide
evidence
that
the
metastasis
suppressor
NM
23
-
H
2
is
involved
in
the
regulation
of
PPAR
δ-mediated
proliferation
.