RNA干涉(RNAi)抑制宫颈癌细胞SiHa中E7基因表达的实验研究

The study of inhibiting the expression of E7 with RNAi in the cervical cancer cell line SiHa

目的探讨HPV16 E7特异性小干涉RNA(siRNA)表达载体对宫颈癌SiHa细胞E7基因的抑制作用。方法构建HPV16 E7特异性表达载体,利用脂质粒转染SiHa细胞,用RT-PCR及Western blot检测其对E7 mRNA及蛋白的影响。结果两条siRNA均能有效抑制E7 mRNA的转录表达,其中以pshRNA-HPV16 E71作用效果最明显,在转染后72h,对E7 mRNA和蛋白的抑制率分别为78.4%和57.6%。结论HPV16 E7 siRNA表达载体对宫颈癌SiHa细胞E7基因的表达有抑制作用。

Objective To test of the effect of vector-based RNA interference （RNAI） technique of inhibiting HPV16E7 gene in SiHa cells of cervical cancer. Methods The HPV16E7-specific siRNA expression vectors siRNA 1, siRNA 2 cere constructed and transfected into SiHa cells byliposone. The expression of HPV16E7 mRNA and protein were detected by real-tie RT-PCR and Western blot. Resuits All the 2 siRNAs could inhibit the level of E7 mRNA significantly, among which siRNA1 was most effective, with an inhibition rates of 78.4 %and57.6 % for the expression of HPV16E7 mRNA and protein respectively 72 hours after transfection. Conclusions Expression carrier of HPV16E7 and siRNa have inhibtional effect on HPV16E7 gene in SiHa cells of cervical cancer.