RNA干扰介导的prohibitin基因沉默对BRL-3A细胞增殖的影响

Effect of RNAi-mediated gene silencing of prohibitin on the proliferation of rat liver cell line BRL-3A

目的观察RNA干扰介导的prohibitin基因沉默对大鼠肝细胞BLR-3A细胞增殖及细胞周期的影响。方法化学合成针对prohibitin基因的小干扰RNA(siRNA),使用脂质体将其转染至BRL-3A细胞中,应用Real-time PCR和Western blot-ting分别检测siRNA转染后细胞prohibitin mRNA和蛋白的表达,MTT法检测转染后的细胞增殖情况,流式细胞术检测转染后细胞周期的变化。结果与对照组比较,prohibitin siRNA转染BRL-3A细胞后,prohibitin mRNA和蛋白表达明显下降;细胞活力升高;G1期细胞减少,S期细胞明显增多(P<0.05)。结论 Prohibitin siRNA能有效抑制大鼠肝细胞BLR-3A细胞prohibitin mRNA和蛋白的表达,并促进细胞增殖。

Objective To investigate the effects of silencing prohibitin gene by small, interfering RNA（siRNA） on the cell prolif- eration and the cell cycle of the rat liver cell line BRL-3A. Methods The siRNAs targeting prihibitin were synthesized,which were transfected into BRL-3A cells using a liposome approach. The expressions of prohibitin mRNA and protein in BRL-3A cells after transfection were determined by quantitative real-time PCR and Western blotting,respectively. The cell proliferation and cell cycle were detected by MTT method and flow cytometry（FCM）, respectively. Results As compared with the control,the expression lev- els of prohibitin mRNA and protein were decreased after transfection with prohibitin siRNA, the cell viability was increased, the number of Gl-phase cells were decreased and S-phase cells were significantly increased（P〈0.05）. Conclusion Prohibitin siRNA can effectively suppress the expressions of prohibitin mRNA and protein in rat liver cell line BRL-3A,and promote the cell prolifer- ation in vitro.