靶向Akt1的siRNA抑制卵巢癌细胞系生长的体外研究  

Growth inhibition on human ovarian carcinoma cells by small interfering RNA targeting Akt1

作　　者：王纪元[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300070 [2]天津市中心妇产科医院妇科,天津300100

出　　处：《现代妇产科进展》2012年第5期345-349,共5页Progress in Obstetrics and Gynecology

摘　　要：目的:探讨靶向Akt1的siRNA抑制人卵巢癌细胞系SKOV3的Akt1表达后,对细胞增殖和侵袭的抑制作用。方法:将Akt1特异性siRNA转染人卵巢癌细胞系SKOV3,Real-time PCR检测Akt1 mRNA表达,Western blot和免疫荧光技术检测Akt1、Akt2、PI3Kp85α和Ki-67的蛋白表达;MTT法和Annexin V标记评价其对细胞增殖活性和凋亡的影响,流式细胞法分析细胞周期变化,Transwell法分析对侵袭能力的影响。结果:转染Akt1 siRNA后可明显下调Akt1 mRNA表达;Akt1、PI3Kp85α、Akt2和Ki-67蛋白的表达水平均明显降低(P<0.01);卵巢癌细胞增殖活性明显降低,细胞周期出现G0/G1阻滞,并诱发细胞凋亡,细胞侵袭能力减弱(P<0.01)。结论:靶向Akt1的siRNA可显著抑制卵巢癌细胞Akt1的表达,进而抑制肿瘤细胞的生长和侵袭,有望成为卵巢癌基因治疗的分子靶点。Objective：To investigate growth inhibition effect on ovarian carcinoma cell line SKOV3 by siRNA targeting Akt1.Methods：RNAi technology was used to observe its inhibitory effect on the growth of human ovarian carcinoma SKOV3 cells.The expressions of Akt1 mRNA were detected by real-time PCR after transfection.The expressions of Akt1,Akt2,PI3Kp85α,Ki-67 were also studied by Western blot and immunofluorescence staining after transfection.MTT methods and annexin V staining were used to evaluate cell proliferation and apoptosis.Flow cytometry was used for cell cycle analysis.Tumor invasion was examined by Transwell analysis.Results：SKOV3 mRNA expression was obviously knocked down after transfection with siRNA.Compared to control and nonsense siRNA transfected cells,SKOV3 cells transfected with siRNA targeting Akt1 showed lowering proliferation activity,while the expressions of PI3Kp85α,Akt1,Akt2,Ki-67 were downregulated.The transfected cells had higher apotosis rate and most cells arresting in the G0/G1 phase.The migration and invasive ability was attenuated.Conclusion：SiRNA targeting Akt1 down regulate expressions of Akt1,inhibit ovarian carcinoma cell proliferation and migration,arrest cell cycle and induce apoptosis in vitro.Consequently,Akt1 can be the promising candidate genes for gene therapy of ovarian cancer.

关 键 词：AKT1 RNAI 卵巢肿瘤 增殖 

分 类 号：R737.31[医药卫生—肿瘤]