沉默线粒体核糖体蛋白L35基因对人食管癌TE-1细胞生长的影响

摘要

Objective:To investigate the effect of lentivirus-mediated silencing of mitochondrial ribosomal protein L35 (MRPL35) gene on the growth of human esophageal cancer TE-1cells, and to clarify its mechanism.Methods:Three kinds of human esophageal cancer cells, TE-1, ECA109and KYSE150, were selected.The relative expression levels of MRPL35mRNA in three kinds of cells by real-time quantitative PCR.The esophageal cancer TE-1cells were divided into shMRPL35group and shCtrl group, and the cells were infected with si-RNA lentivirus and si-RNA lentivirus;the esophageal cancer cell line stably silenting the MRPL35gene was established.Real-time quantitative PCR and Western blotting methods were used to detect the efficiency of MRPL35gene silencing.The cell growth curves in various groups were detected by CCK-8method, and the apoptotic rates were detected by flow cytometry after AnnexinⅤ-PE/7AAD double staining.Results:Three kinds of esophageal cancer cells expressed MRPL35gene, and the expression levels were not statistically significant between them (P＞0.05) .The results of real-time quantitative PCR and Western blotting methods showed that the mRNA and protein levels of MRPL35in the TE-1cells in shMRPL35group were significantly lower than those in shCtrl group (P＜0.05) .Compared with shCtrl group, the cell growth speed in shMRPL35group was decreased (P＜0.05) , and the apoptotic rate was significantly increased (P＜0.01) .Conclusion:Silencing MRPL35gene can inhibit the proliferation of esophageal cancer TE-1cells and plays a role through the apoptotic pathway.%目的:探讨慢病毒介导沉默线粒体核糖体蛋白L35 (MRPL35) 基因对人食管癌TE-1细胞生长的影响, 并阐明其机制.方法:选择3种人食管癌TE-1、ECA109和KYSE150细胞, 采用实时定量PCR法检测3种细胞中MRPL35mRNA相对表达水平.食管癌TE-1细胞分为shMRPL35组和shCtrl组, 分别加入沉默靶基因带有嘌呤霉素抗性的si-RNA慢病毒和带有嘌呤霉素抗性的阴性对照si-RNA慢病毒进行感染, 建立稳定沉默MRPL35基因的食管癌细胞系, 实时定量PCR及Western blotting法检测各组细胞中MRPL35基因沉默效率, 采用CCK-8法检测各组细胞生长曲线, AnnexinⅤ-PE/7AAD双染色后流式细胞术检测细胞凋亡率.结果:3种食管癌细胞均表达MRPL35基因, 3种细胞中MRPL35mRNA表达水平比较差异无统计学意义 (P＞0.05) .实时定量PCR法和Western blotting法检测, shMRPL35组TE-1细胞中MRPL35mRNA和蛋白表达水平明显低于shCtrl组 (P＜0.05) .与shCtrl组比较, shMRPL35组TE-1细胞生长速度明显降低 (P＜0.05) , 凋亡率明显升高 (P＜0.01) .结论:沉默MRPL35基因可抑制食管癌细胞TE-1增殖, 并通过凋亡途径发挥作用.