靶向PI3Kp85α的siRNA抑制人卵巢癌细胞系生长的实验研究  

Inhibitory Effects of Small Interfering RNA Targeting on the Growth of Human Ovarian Carcinoma Cells Phosphoinositide 3-kinase Regulatory Subunit p85 alpha

作　　者：郭祥翠[1] 朱颖军[2] 林琬君[2] 

机构地区：[1]天津医科大学研究生院,天津市300070 [2]天津市中心妇产科医院

出　　处：《中国肿瘤临床》2012年第7期369-372,共4页Chinese Journal of Clinical Oncology

摘　　要：目的:探讨siRNA靶向抑制卵巢癌细胞磷脂酰肌醇3-激酶调节亚基p85α(phosphoinositide 3-kinase regulatory subunitp85 alpha,P13Kp85α)表达后,对肿瘤细胞增殖和侵袭的影响。方法:应用siRNA靶向PI3Kp85α转染人卵巢癌细胞系SKOV3,Re-altime PCR检测目的基因mRNA表达,Western blot和免疫荧光技术检测PI3Kp85α、AKT1、AKT2和Ki67的表达变化;M_rr法和an-nexin V标记评价细胞的增殖活性和凋亡,流式细胞法分析细胞周期.Transwell法分析侵袭能力。结果:转染PI3Kp85αsiRNA后PI3Kp85αmRNA表达下降;PI3Kp85α、AKT1、AKT2和Ki67的蛋白表达水平均明显降低(P<0.01)。卵巢癌细胞增殖活性明显受到抑制,细胞周期在G_0/G_1期有明显的阻滞,早期凋亡的细胞教明显增多,细胞的运动迁移能力降低(P<0.01),侵袭能力减弱(P<0.01)。结论:靶向PI3Kp85α的siRNA技术可抑制卵巢癌细胞PI3Kp85α的表达,进而抑制肿瘤细胞的生长,这可能成为卵巢癌基因治疗的新靶点。Objective： This study investigates the growth inhibition effect of small interfering RNA （ siRNA ） targeting Phos- phoinositide 3-kinase Regulatory Subunit p85 alpha （PI3Kp85α） on ovarian carcinoma cells. Methods： RNA interference （ RNAi ） tech- nology was used to observe the inhibitory effect of siRNA on the growth of human ovarian carcinoma SKOV3 cells. The expression of PI3Kp85α mRNA was detected through real-time polymerase chain reaction after the transfection. The expression of PI3Kp85α, AKT1, AKT2, and Ki-67 was also studied using Western blot and immunofluorescence staining after transfection. Moreover, MTT methods and annexin V staining were used to evaluate cell proliferation and apoptosis. Flow cytometry was used for cell cycle analysis, and tu- mor invasion was examined through Transwell analysis. Results： SKOV3 PI3Kp85α mRNA expression was significantly knocked down after transfection with siRNA. The SKOV3 cells transfected with siRNA targeting PI3Kp85α showed lower proliferation activity, whereas the expression of PI3Kp85α, AKT1, AKT2, and Ki-67 was downregulated compared with the control and nonsense siRNA transfected cells. The transfected cells had higher apoptosis rate and most cells arrested in the G0/G1 phase. Moreover, the migration and invasive ability of the ovarian carcinoma cells was attenuated. Conclusion： Silencing PI3Kp85α genes with siRNA downregulated the mRNA expression of PI3Kp85α in ovarian carcinoma endothelial cells, blocked cell proliferation and migration, arrested cell cycle, and induced apoptosis in vitro. Therefore, P13Kp85α can be a candidate gene for ovarian cancer gene therapy.

关 键 词：PI3Kp85α RNAI 卵巢癌 增殖 

分 类 号：R737.31[医药卫生—肿瘤]