载体介导的RNAi技术抑制宫颈癌CaSki细胞E7基因的表达

Inhibiting E7 Oncogene in CaSki Cells of Cervical Cancer by Vector-based RNA Interference Technique

目的探讨H PV 16E 7特异性小干扰RNA(s iRNA)表达载体对宫颈癌C aSk i细胞E 7基因的抑制作用。方法构建H PV 16 E 7特异性s iRNA表达载体,利用脂质体转染C aSk i细胞,以荧光定量RT-PCR及W esternb lot检测其对E 7 mRNA及蛋白表达的影响。结果3种表达载体P 1、P 2、P 3均能抑制C aSk i细胞E 7基因mRNA和蛋白的表达,其中载体P 1抑制作用最强,在转染后3周,对E 7 mRNA和蛋白的抑制率分别为92.86%和81.0%。结论H PV 16E 7 s iRNA表达载体能有效地抑制宫颈癌C aSk i细胞E 7基因的表达。

Objective To test of the effect of vector-based RNA interference （RNAi） technique on inhibiting HPV16E7 gene in CaSki cells of cervical cancer. Methods The HPV16E7-specific siRNA expression vectors P1, P2 and P3 were constructed and transfected into CaSki cells by liposome. The expression of HPV16E7 mRNA and protein were detected by real-time RT-PCR and Western blot. Results The expression of HPV16E7 mRNA and protein decreased with the transfection of P1, P2 and P3. Vector P1 had the strongest inhibition effect, with an inhibition rates of 92. 86% and 81. 0% for the expression of HPV16E7 mRNA and protein respectively three weeks after transfection. Conclusion The expression of E7 gene in CaSki cells can be inhibited by HPV16E7-specific siRNA expression vector.