siRNA表达载体抑制宫颈癌SiHa细胞增殖的实验研究

Experimental Study on Inhibition of Cervical Carcinoma SiHa Cell Proliferation by siRNA Expression Vector

目的观察HPV16E7 siRNA表达载体对宫颈癌SiHa细胞增殖的影响。方法利用脂质体将HPV16E7特异性siRNA表达载体转染SiHa细胞,通过荧光定量RT-PCR检测E7 mRNA的变化;流式细胞仪检测E7蛋白的变化和细胞周期的分布;MTT法检测细胞增殖的变化。结果HPV16E7 siRNA表达载体可明显抑制SiHa细胞E7 mRNA和E7蛋白的表达,抑制率分别为92.15%、84.30%;阻滞SiHa细胞由G1期进入S期;抑制细胞的生长。结论HPV16E7 siRNA表达载体可以特异高效地抑制宫颈癌SiHa细胞E7基因的表达,从而调控肿瘤细胞周期,抑制肿瘤细胞增殖。

Objective To observe the effect of HPV16E7 specific expression vector on cell proliferation in cervical carcinoma SiHa cells. Methods The HPV16E7 siRNA expression vector and empty expression vector were transfected into SiHa cells by liposome. The effects on E7 mRNA and E7 protein expression, cell cycle phase and cell growth rate were examined respectively by real-time RT-PCR, FCM and MTT assay. Results The HPV16E7 siRNA expression vector significantly inhibited the expression levels of E7 mRNA and E7 protein, the inhibition rates being 92.15% and 84.30% respectively. It also inhibited the transition from G1 phase to S phase and the growth of SiHa ceil llne. Conclusion HPV1E7 specific siRNA expression vector could specifically and efficiently inhibit the expression of E7 gene and hence it could regulate cell cycle and inhibit cell proliferation in cervical carcinoma SiHa cells.