目的：建立慢病毒介导RhoA基因稳定沉默卵巢癌HO8910细胞株，研究RhoA基因对HO8910细胞侵袭和迁移的影响。方法：构建靶向沉默RhoA基因的慢病毒载体Lenti-U6-shRhoA，将其感染HO8910细胞，经过嘌呤霉素筛选，建立稳定沉默RhoA基因的HO8910细胞。实时荧光定量PCR检测HO8910细胞中RhoA mRNA的表达；MTT检测HO8910细胞的增殖；Transwell体外迁移和侵袭实验检测RhoA基因沉默对HO8910细胞侵袭和迁移的影响。结果：成功构建靶向RhoA基因的慢病毒载体Lenti-U6-shRhoA，建立了RhoA基因稳定沉默的HO8910细胞。与Lenti-U6-NC对照组对比， RhoA基因稳定沉默HO8910细胞RhoA mRNA的表达明显降低\[（30.7±5.40）% vs（95.9±6.53）%，P<0.05\]；沉默RhoA基因能降低HO8910细胞的存活率\[（51.16±7.41）% vs（95.67 ±2.14）%，P<0.05\]；RhoA基因沉默组HO8910细胞的侵袭、迁移及黏附能力较Lenti-U6-NC对照组明显受到抑制\[迁移穿过人工基底膜细胞数（31±6） vs （84±13）个，P<0.05；穿过微孔膜的细胞数(97±12) vs (689±23)个，P<0.05；黏附率 (68.16±6.53)% vs (98.71±4.92)%，P<0.05\]。结论：慢病毒介导的RhoA基因沉默能抑制卵巢癌HO8910细胞的侵袭和迁移。

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 (\[51.16±741\]% vs \[95.67±214\]%, 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 \[9871±492\]%, P<005) 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.

广东省科技计划资助项目（No. 2010B031600185）