慢病毒靶向携带Akt1 shRNA对胃癌细胞SGC-7901凋亡和增殖的影响  

Effect of lentivirus targeted to bring Akt1 shRNA on apoptosis and proliferation of gastric carcinoma SGC-7901 Cells

作　　者：谭韡[1] 于红刚[1] 罗和生[1] 周巍[1] TAN Wei YU Honggang LUO Hesheng ZHOU Wei(Department of Gastroenterology, Renmin Hospital of Wuhan University Hubei Key Laboratory of Digestive System Disease, Hubei Province, Wuhan 430060, Chin)

机构地区：[1]武汉大学人民医院消化内科消化系统疾病湖北省重点实验室,湖北武汉430060

出　　处：《中国医药导报》2017年第17期23-27,共5页China Medical Herald

基　　金：国家自然科学基金资助项目(81401959)

摘　　要：目的探讨慢病毒载体靶向携带Akt1基因短发夹RNA(Akt1-shRNA)对胃癌细胞SGC-7901凋亡和增殖的影响。方法构建表达Akt1-shRNA的慢病毒载体,转染293T细胞后获得Akt1-shRNA的慢病毒颗粒,病毒感染胃癌SGC-7901细胞(SGC-LV-shAkt1)为实验组细胞,SGC7901细胞和转染慢病毒空载体SGC7901细胞(SGCLV-sh CON)为对照组细胞。Real-Time PCR检测Akt1 m RNA表达水平,Western blot检测胃癌细胞Akt1、phosphoAkt(ser473)和bcl-2蛋白表达水平,然后流式双染法、生长速率等方法检测Akt1 shRNA对SGC-7901细胞凋亡及增殖能力的影响。结果成功构建Akt1基因RNAi的慢病毒载体LV-shAkt1,病毒滴度为5×108 TU/mL。SGC-7901细胞转染后,实验组(SGC-LV-shAkt1)Akt1 m RNA和Akt1、p-Akt、bcl-2蛋白表达水平较对照组(SGC-7901、SGC-LV-sh CON)降低(P<0.01);(38.7±3.5)%的实验组细胞凋亡(P<0.05)。SGC-LV-shAkt1细胞生长曲线平缓,生长速度较另两种细胞(SGC-7901、SGC-LV-sh CON)明显降低。结论靶向Akt1的RNAi能有效抑制胃癌细胞SGC-7901的生长,诱导胃癌细胞凋亡,其机制可能与抑制PI3K/Akt信号通路,下调抗凋亡蛋白bcl-2有关。Objective To investigate the apoptosis and proliferation effect of lentiviral vectors shRNA for Akt1 to human gastric carcinoma SGC-7901 cells.Methods The lentiviral vectors Akt1-shRNA were constructed, lentivirus Akt1-shRNA was transfected into human gastric carcinoma SGC-7901 cells, the SGC-7901 cell strained with Akt1 gene silencing perpetually（SGC-LV-sh CON） was as conrol group cell and amplified（SGC LV-shAkt1） was as experiment group cell.Effect of Akt1 on SGC-7901 cells were measured by RT-PCR, Western-blot, flow cytometry of annexin V-FITC/PI, tumor growth curve.Results Akt1 RNAi lentiviral vectors LV-shAkt1 was established successfully,virus titer was 5 ×108TU/mL.After SGC-7901 cell transfection, the expression of Akt1 m RNA in experiment group（SGC-LV-shAkt1） was weaker than that in the control group（SGC-7901, SGC-LV-sh CON）, and Akt1, p-Akt and bcl-2 protein expressions in experiment group was inhabited（P 〈0.01）,（38.7±3.5）% cell in experiment group happened apotosis（P〈 0.05）.The cell growth curve of SGC-LV-shAkt1 cell was gentle, compared with SGC-7901 cell or SGCLV-sh CON cell, the growth was lower.Conclusion The lentiviral vectors mediated shRNA for Akt1 can significantly inhibit the expression of Akt1 gene in vitro and markedly inhibit SGC-7901 cell growth and induce the apoptosis of gastric carcinoma cell.The mechanism may be down-regulation of PI3K/Akt signal pathway and anti-apoptosis protein bcl-2 expression.

关 键 词：AKT1 慢病毒载体 胃癌 凋亡 

分 类 号：R735.2[医药卫生—肿瘤]