Bmi-1基因RNAi对胃癌细胞SGC7901及AGS作用的初步研究

Effects of RNAi of Bmi-1 gene on gastric carcinoma SGC7901 and AGS cells

目的:构建Bmi-1基因的RNAi表达载体,观察其对胃癌细胞SGC7901与AGS增殖、侵袭能力的影响。方法:利用慢病毒表达体系pHelper1.0/pHelper2.0/pGCL-GFP,构建Bmi-1基因的RNAi重组质粒vshRNA-Bmi-1。适时定量PCR和蛋白质印迹法分别检测稳定转染vshRNA-Bmi-1后胃癌细胞中Bmi-1 mRNA及蛋白的表达;MTT法及小室侵袭实验检测胃癌细胞增殖和侵袭能力的变化。结果:稳定转染vshRNA-Bmi-1后SGC7901及AGS细胞Bmi-1 mRNA表达均明显下降,抑制率分别为85%及80%。SGC7901细胞中Bmi-1蛋白表达无明显变化,而在AGS细胞中明显下降。Bmi-1干扰后SGC7901细胞的生长及侵袭能力无明显变化,而AGS细胞增殖减慢、侵袭能力减弱。结论:成功构建Bmi-1基因的RNAi表达载体,vshRNA-Bmi-1重组质粒明显下调Bmi-1 mR-NA在胃癌细胞SGC7901和AGS中的表达。Bmi-1基因的RNAi能抑制胃癌细胞AGS的增殖和侵袭能力。

OBJECTIVE：To construct the Bmi1 RNAi expression vector and investigate its influence on the proliferation and invasiveness of gastric carcinoma SGC7901 and AGS cells.METHODS： The recombinant vshRNA-Bmi-1 plasmid of Bmi1 RNAi was constructed by the lentiviral expression system,pHelper1.0/pHelper2.0/pGCL-GFP.Bmi-1 mRNA and protein expressions of stably transfected gastric carcinoma cells were identified respectively by real-time PCR and Western blot analyses and the changes of the proliferation and invasiveness of gastric carcinoma were detected by MTT and an invasion assay.RESULTS：The Bmi-1 mRNA expressions of stably transfected gastric carcinoma SGC7901 and AGS cells were significantly decreased with inhibitory rates of 85% and 80% respectively.The expression of Bmi-1 protein in gastric carcinoma AGS cells was significantly decreased,whereas there was no significant difference in SGC7901 cells.The proliferation and invasiveness of AGS cells significantly reduced but no significant changes were observed in SGC-7901 cells.CONCLUSION： Successful construction of the Bmi1 RNAi expression vector with reduced Bmi-1 expression of AGS cells demonstrates the role of Bmi-1 RNAi in restraining proliferation and invasiveness of gastric carcinoma cells.