Silencing heparanase suppresses proliferation and invasion in ovarian cancer SKOV3 cells
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Silencing heparanase suppresses proliferation and invasion in ovarian cancer SKOV3 cells
Journal of Army Medical UniversityVol. 33, Issue 22, Pages: 2361-2365(2011)
作者机构:
第三军医大学新桥医院妇产科
作者简介:
基金信息:
DOI:
CLC:R394.3;R73-362;R737.31
Published:2011,
Published Online:30 November 2011,
稿件说明:
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ZHAO LING, YANG YING. Silencing heparanase suppresses proliferation and invasion in ovarian cancer SKOV3 cells. [J]. Journal of army medical university, 2011, 33(22): 2361-2365.
DOI:
ZHAO LING, YANG YING. Silencing heparanase suppresses proliferation and invasion in ovarian cancer SKOV3 cells. [J]. Journal of army medical university, 2011, 33(22): 2361-2365.DOI:
Silencing heparanase suppresses proliferation and invasion in ovarian cancer SKOV3 cells
Objective To explore the effect of silencing heparanase gene by RNA short hairpin on the proliferation and invasion in human ovarian cancer cell line SKOV3. Methods After shRNA lentiviral vectors targeting heparanase gene was constructed
they were transfected into SKOV3 cells. Experimental transfection groups included the interference sequence
s HPA shRNA-1 group
HPA shRNA-2
and HPA shRNA-3 group
and the untransfected group amd blank transfection group served as control. After transfection
the interference efficiency was observed though fluorescence quantitative PCR and Western blotting to detect the expression of heparanase at mRNA and protein levels. Flow cytometry was employed to test cell cycle changes
cell counting kit-8 (CCK-8) for the detection of cell proliferation
and matrix gel invasion assays for the detection of cell invasion ability. Results After shRNA lentiviral vectors were transfected into SKOV3 cells
HPA expression was significantly decreased at mRNA and protein levels in the HPA-shRNA-1 sequence and the HPA-shRNA-2 sequence groups (
P
<
0.05)
whereas the expression levels of HPA-shRNA-3 sequence group was not decreased
indicating it was an ineffective sequence. In the SKOV3 cells transfected with effective sequences HPA-shRNA-1 and HPA-shRNA-2
the cell percentage of G
1
phase was decreased
the proliferation (OD index) was significantly decreased and transmembrane cell number was increasingly decreased. There was a significantly difference between the experimental groups and untransfected and blank transfection groups respectively (
P
<
0.05). Conclusion Silencing HPA by RNA interference inhibits heparanase expression
and suppresses cell proliferation and invasion efficiently
which may be correlated with the downregulation of HPA gene and protein.
Role of C3G/Rap1 and Dock180/Rac1 signal pathway in infiltration of ovarian cancer
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