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Function of RasGRP3 in the formation and progression of human breast cancer.
[severe combined immunodeficiency]
Ras
guanine
nucleotide
exchange
factors
(
RasGEFs
)
mediate
the
activation
of
the
Ras
signaling
pathway
that
is
over
activated
in
many
human
cancers
.
The
RasGRP
3
,
an
activator
of
H-Ras
and
R-Ras
protein
exerts
oncogenic
effects
and
the
overexpression
of
the
protein
is
observed
in
numerous
malignant
cancer
types
.
Here
,
we
investigated
the
putative
alteration
of
expression
and
potential
function
of
RasGRP
3
in
the
formation
and
progression
of
human
breast
cancer
.
The
RasGRP
3
and
phosphoRasGRP
3
expressions
were
examined
in
human
invasive
ductal
adenocarcinoma
derived
samples
and
cell
lines
(
BT
-
474
,
JIMT-
1
,
MCF
7
,
SK-
BR
-
3
,
MDA-
MB
-
453
,
T
-
47
D
)
both
in
mRNA
(
Q-PCR
)
and
protein
(
Western
blot
;
immunohistochemistry
)
levels
.
To
explore
the
biological
function
of
the
protein
,
RasGRP
3
knockdown
cultures
were
established
.
To
assess
the
role
of
RasGRP
3
in
the
viability
of
cells
,
annexin-
V
/
PI
staining
and
MitoProbe
â„¢
DilC
1
(
5
)
assay
were
performed
.
To
clarify
the
function
of
the
protein
in
cell
proliferation
and
in
the
development
of
chemotherapeutic
resistance
,
CyQuant
assay
was
performed
.
To
observe
the
RasGRP
3
function
in
tumor
formation
,
the
Severe
combined
immunodeficiency
(
SCID
)
mouse
model
was
used
.
To
investigate
the
role
of
the
protein
in
Ras-related
signaling
Q-PCR
and
Western
blot
experiments
were
performed
.
R
asGRP
3
expression
was
elevated
in
human
breast
tumor
tissue
samples
as
well
as
in
multiple
human
breast
cancer
cell
lines
.
Down-regulation
of
RasGRP
3
expression
in
breast
cancer
cells
decreased
cell
proliferation
,
induced
apoptosis
in
MCF
7
cells
,
and
sensitized
T
-
47
D
cells
to
the
action
of
drugs
Tamoxifen
and
trastuzumab
(
Herceptin
)
.
Gene
silencing
of
RasGRP
3
reduced
tumor
formation
in
mouse
xenografts
as
well
.
Inhibition
of
RasGRP
3
expression
also
reduced
Akt
,
ERK
1
/
2
and
estrogen
receptor
alpha
phosphorylation
downstream
from
IGF-
I
insulin
like
growth
factor
-
I
(
IGF-
I
)
or
epidermal
growth
factor
(
EGF
)
stimulation
confirming
the
functional
role
of
RasGRP
3
in
the
altered
behavior
of
these
cells
.
Taken
together
,
our
results
suggest
that
the
Ras
activator
RasGRP
3
may
have
a
role
in
the
pathological
behavior
of
breast
cancer
cells
and
may
constitute
a
therapeutic
target
for
human
breast
cancer
.
Diseases
Validation
Diseases presenting
"growth factor"
symptom
22q11.2 deletion syndrome
achondroplasia
adrenal incidentaloma
aniridia
cadasil
cholangiocarcinoma
coats disease
dedifferentiated liposarcoma
dentin dysplasia
dentinogenesis imperfecta
dystrophic epidermolysis bullosa
esophageal carcinoma
esophageal squamous cell carcinoma
hodgkin lymphoma, classical
holt-oram syndrome
inclusion body myositis
kallmann syndrome
krabbe disease
liposarcoma
lymphangioleiomyomatosis
oculocutaneous albinism
oral submucous fibrosis
pleomorphic liposarcoma
primary effusion lymphoma
primary hyperoxaluria type 1
severe combined immunodeficiency
systemic capillary leak syndrome
von hippel-lindau disease
wolf-hirschhorn syndrome
x-linked adrenoleukodystrophy
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