Bmi-1-siRNA对食管癌细胞株EC9706的影响

Effects of Bmi-1-siRNA on human esophageal EC9706 cells

摘要:

目的：探讨Bmi-1-siRNA对食管癌EC9706细胞的影响。方法：将不同浓度（50、100、150 nmol/L）的Bmi-1-siRNA转染EC9706细胞24、48、72 h，MTT法检测各组各时间段细胞增殖情况，免疫细胞化学检测转染后EC9706细胞内Bmi-1、端粒酶逆转录酶（human telomerase reverse transcriptase，hTERT）、P16蛋白的表达，原位杂交方法检测Bmi-1 mRNA的表达。结果：Bmi-1-siRNA对EC9706细胞的生长抑制率明显升高，且具有时间依赖性和剂量依赖性（F=1.322、2.881，P＜0.001）；EC9706细胞中Bmi-1、hTERT蛋白表达明显降低，P16蛋白表达升高（F=12.229、23.556、19.414，P＜0.001）。结论：Bmi-1-siRNA有效抑制EC9706细胞的增殖，其机制可能为降低Bmi-1 mRNA及Bmi-1、hTERT蛋白的表达，上调P16蛋白的表达。

Abstract:

Objective：To explore the effects of Bmi-1-siRNA on human esophageal EC9706 cells. Methods：Different concentrations of Bmi-1-siRNA（50，100，150 nmol/L） was used to transfect EC9706 with different time exposures（24，48，72 h）. Changes of cell cycle in EC9706 cells before and after the transfection were detected by MTT. Levels of protein of Bmi-1，telomerase reverse transcrip－tase（hTERT），P16 in transfected cells were measured by immunocytochemistry. Levels of Bmi-1mRNA in transfected cells were measured by situ hybridization. Results：Bmi-1-siRNA effectively inhibited cell proliferation of EC9706 cells in a dose and time dependent way（F=1.322，2.881；P<0.001）. Expression of Bmi-1 and hTERT protein was decreased and that of P16 was increased（F=12.229，23.556，19.414；P<0.001）. Conclusions：RNAi of Bmi-1 in EC9706 cell line could effectively inhibit EC9706 cell proliferation，whose mechanism may relate with down-regulation of Bmi-1 mRNA，Bmi-1，hTERT protein expression and up-regulation of P16 protein expression.