RNA干扰survivin对卵巢癌HO-8910PM细胞周期的影响

目的 探讨RNA干扰survivin基因对卵巢癌HO-8910PM细胞周期的影响.方法 实验分3个组：siR-NA组,采用脂质体法将靶向survivin siRNA荧光真核表达质粒载体pGPU6-GFP-survivin-shRNA转染卵巢癌HO-8910PM细胞;空白对照组,单纯细胞培养;阴性对照组,用表达与survivin序列无同源性的siRNA片段质粒pGPU6-GFP-NC转染细胞.于转染后48 h和72 h,用PI单染法流式细胞术检测各组HO-8910PM细胞周期的变化以及Western blot检测survivin蛋白的表达情况.结果 PI单染流式细胞仪检测细胞周期结果表明,与空白对照组和阴性对照组相比较,2个时间点siRNA组转染后的G0/G1期细胞比例明显增加,而G2/M期细胞比例均下降（P＜ 0.05）;Western blot结果证实,siRNA组的HO-8910PM细胞survivin蛋白的表达量明显下调.结论 导入survivin基因特异性的siRNA可导致卵巢癌HO-8910PM细胞周期阻滞于G0/G1期,细胞增殖受阻,且细胞sur-vivin蛋白表达量下降.

Effect of survivin gene silence on cell cycle of HO-8910PM ovarian cancer cells

Objective To explore the effect of RNA interfered silence of the survivin geng on ovarian cancer HO- 8910PM cell cycle. Methods Experiments were set up three groups： siRNA group, lipo- some targeting survivin siRNA eukaryotic expression vector pGPU6 - GFP - survivin - shRNA transfeet- ed ovarian cancer HO- 8910PM cells; Blank control group with cultured cells alone ; Negative control group, with the expression of survivin siRNA sequence but？ no homology pGPU6 - GFP - NC plasmid transfected cells. Forty- eighth and 72h after transfection, the HO - 8910PM cell cycle changes were detected by PI staining flow cytometry method, and the survivin protein expression was detected by Western blot. Results Compared with the blank control group and negative control group, PI staining flow cytometry results showed that after transfection of siRNA GO/G 1 phase cells increased , while the G 2/M phase ceils decreased （ P 〈 0.05 ） ; Western blot analysis confirmed that in siRNA of HO - 8910PM cells, survivin protein expression was significantly reduced. Conclusion Importing survivin gene -specific siRNA can lead to ovarian cancer HO -8910PM cell cycle arrest at GO/G1 phase, cell proliferation blocked, and cellular survivin protein expression decreased.