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2015 ; 12
(2
): 1783-8
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Effect of the WWOX gene on the regulation of the cell cycle and apoptosis in
human ovarian cancer stem cells
#MMPMID25891642
Yan H
; Tong J
; Lin X
; Han Q
; Huang H
Mol Med Rep
2015[Aug]; 12
(2
): 1783-8
PMID25891642
show ga
In order to examine new ideas for gene therapy in ovarian cancer, the specific
mechanism underlying the effects of the WW domain containing oxidoreductase
(WWOX) gene on cell cycle regulation and apoptosis in human ovarian cancer stem
cells was investigated. Ovarian cancer stem cells were transfected with a
eukaryotic expression vector carrying the WWOX gene in vitro (recombinant
plasmid) and cells transfected with the empty plasmid (empty plasmid) or
untransfected cells were used as controls. Stably transfected cells were screened
and amplified in culture and the WWOX protein was detected by western blot
analysis in the three groups of cells. Western blot analysis was performed to
detect the expression of cell cycle regulatory proteins cyclin E,
cyclin-dependent kinase (CDK) 2, cyclin D1, CDK4 and apoptosis-related protein
Wnt-5? and c-Jun N-terminal kinase (JNK), while polymerase chain reaction (PCR)
was used to detect alterations in the mRNA expression levels of caspase-3. The
results demonstrated that the WWOX protein was stably expressed in cells of the
recombinant plasmid group, but was not detected in cells of the empty plasmid
group and the control group. Cell proliferation at each time point decreased
significantly in the recombinant plasmid group compared with the empty plasmid
group and the control group. Flow cytometric analysis demonstrated that the
proportion of cells in the G0/G1 phase in the recombinant plasmid group was
significantly higher than that of cells in the empty plasmid group and the
control group. The rate of apoptosis in the recombinant plasmid group was
significantly higher than that of cells in the empty plasmid group and the
control group. Western blot analysis demonstrated that the expression levels of
cyclin E, CDK2, cyclin D1 and CDK4 in the recombinant plasmid group were
significantly lower than those in the empty plasmid group and the control group;
however, the expression levels of Wnt-5? and JNK were significantly higher than
those in the empty plasmid group and the control group. PCR results demonstrated
that the mRNA expression level of caspase-3 in the recombinant plasmid group was
significantly higher than that in the empty plasmid group and the control group.
In conclusion, the present study demonstrated that the WWOX gene can be stably
expressed in ovarian cancer stem cells and that it inhibits the proliferation of
ovarian cancer stem cells. The WWOX gene can downregulate the expression levels
of cell cycle proteins cyclin E-CDK2 and cyclin D1-CDK4, which affects the cell
cycle of ovarian cancer stem cells. Furthermore, the WWOX gene can upregulate the
mRNA expression levels of Wnt-5?, JNK and caspase-3, thus contributing to
apoptosis of ovarian cancer stem cells. The present study demonstrated that the
WWOX gene may be an important molecular target for the treatment of ovarian
cancer in the future.
|*Apoptosis
[MESH]
|Caspase 3/genetics/metabolism
[MESH]
|Cell Line, Tumor
[MESH]
|Cell Proliferation
[MESH]
|Cyclin D1/metabolism
[MESH]
|Cyclin E/metabolism
[MESH]
|Cyclin-Dependent Kinase 2/metabolism
[MESH]
|Cyclin-Dependent Kinase 4/metabolism
[MESH]
|Down-Regulation
[MESH]
|Female
[MESH]
|G1 Phase Cell Cycle Checkpoints
[MESH]
|Humans
[MESH]
|JNK Mitogen-Activated Protein Kinases/genetics/metabolism
[MESH]
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