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

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MRPL35 mRNA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，Annexin Ⅴ-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MRPL35 mRNA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MRPL35 mRNA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。

Effect of silencing mitochondrial ribosomal protein L35 gene on growth of human esophageal cancer TE-1 cells

Objective: To investigate the effect of lentivirus-mediated silencing of mitochondrial ribosomal protein L35 (MRPL35) gene on the growth of human esophageal cancer TE-1 cells,and to clarify its mechanism.Methods: Three kinds of human esophageal cancer cells, TE-1, ECA109 and KYSE150, were selected. The relative expression levels of MRPL35 mRNA in three kinds of cells by real-time quantitative PCR. The esophageal cancer TE-1 cells were divided into shMRPL35 group and shCtrl group,and the cells were infected with si-RNA lentivirus and si-RNA lentivirus; the esophageal cancer cell line stably silenting the MRPL35 gene was established. Real-time quantitative PCR and Western blotting methods were used to detect the efficiency of MRPL35 gene silencing.The cell growth curves in various groups were detected by CCK-8 method, and the apoptotic rates were detected by flow cytometry after Annexin Ⅴ-PE/7AAD double staining.Results: Three kinds of esophageal cancer cells expressed MRPL35 gene, 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 MRPL35 in the TE-1 cells in shMRPL35 group were significantly lower than those in shCtrl group(P<0.05). Compared with shCtrl group, the cell growth speed in shMRPL35 group was decreased (P<0.05), and the apoptotic rate was significantly increased (P<0.01).Conclusion: Silencing MRPL35 gene can inhibit the proliferation of esophageal cancer TE-1cells and plays a role through the apoptotic pathway.