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Effect of lentivirus-mediated silence of RhoA gene on invasion and migration of ovarian cancer cells
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Abstract:
Objective:To construct a ovary cancer cell line HO8910 with a stable knockdown of RhoA gene by lentiviral vectors; and to investigate the effect of RhoA gene on invasion and migration of HO8910 cells. Methods: Lenti-U6-shRhoA lentivirus targeting silence RhoA gene expression was constructed and infected into HO8910 cells. HO8910 cells stable knockdown RhoA gene expression were obtained by puromycin selection. The expression of RhoA mRNA in HO8910 cells was detected by real-time PCR; the proliferation of HO8910 cells was detected by MTT assay; and Transwell migration assay and Matrigel invasion assay were used to measure the effect of RhoA gene silence on invasion and migration of HO8910 cells. Results: The lentiviral vector Lenti-U6-shRhoA targeting RhoA gene was constructed successfully and a stable RhoA knockdown HO8910 cell line was established. The RhoA gene expression was efficiently suppressed by Lenti-U6-shRhoA with a expression rate of (30.7±5.4)%, compared with the control Lenti-U6-NC group (95.9±6.53)% (P<0.05). The knockdown of RhoA gene inhibited the survival rate of HO8910 cells (\[51.16±741\]% vs \[95.67±214\]%, P<0.05); Compared with the Lenti-U6-NC group, the capabilities of the invasion (97±12 vs 689±23, P<0.05), migration (31±6 vs 84±13, P<0.05) and adhesion rate (\[68.16±6.53\]% vs \[9871±492\]%, P<005) were also obviously suppressed in the RhoA stable knockdown HO8910 cell line. Conclusion: The lentivirus-mediated RhoA gene silence can suppress the invasion and migration of ovary cancer HO8910 cells.
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Project Supported:
Project supported by the Science and Technology Program of Guangdong Province (No. 2010B031600185)
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