HPV16E7 siRNA表达载体抑制宫颈癌SiHa细胞E7基因的研究

目的：研究HPV16E7siRNA表达载体对宫颈癌SiHa细胞E7基因的抑制作用。方法：利用脂质体将HPV16E7 siRNA表达载体psiRNA-1、psiRNA-2、psiRNA-3及空载体psiRNA转染SiHa细胞,以荧光定量RT-PCR和流式细胞仪检测不同时间点E7 mRNA和蛋白的变化。结果：载体psiRNA-1、psiRNA-2和psiRNA-3均能抑制SiHa细胞E7基因 mRNA和蛋白的表达,其中载体psiRNA-1的抑制作用最强。在抗性克隆形成后1周和4周,对E7mR-NA和蛋白抑制率分别为92.15%、84.30%和65.69%、59.11%。而空载体对E7mR-NA和蛋白的表达均无明显抑制作用。结论：HPV16E7 siRNA表达载体能较长期地抑制宫颈癌SiHa细胞E7基因的表达。

Inhibition of E7 oncogene in SiHa cells of cervical cancer by HPV16E7 specific siRNA expression vector

OBJECTIVE:To investigate the inhibition of E7 gene in SiHa cells of cervical cancer by HPV16E7 specific siRNA expression vector in vitro.METHODS:HPV16E7 specific siRNA expression vectors psiRNA-1,psiRNA-2,psiRNA-3 and empty vectcor were transfected into SiHa cells by liposome. The expression levels of HPV16E7 mRNA and protein were detected by real-time RT-PCR and flow cytometry at different time after transfection.RESULTS:The expressions of HPV16E7 mRNA and protein were inhibited by vector psiRNA-1,psiRNA-2,psiRNA-3. Among them the inhibition effect of vector psiRNA-1 was the most effective and the inhibition rates were 92.15%, 84.30% and 65.69%, 59.11% respectively at one week and four weeks after the formation of SiHa resistant clone. However, the empty vector had no significant inhibition effect.CONCUSION:The expression of E7 gene in SiHa cells can be inhibited by HPV16E7 siRNA expression vector for a relatively long time.