RNAi表达载体对宫颈癌HPV16E6基因表达的抑制作用

The Inhibitory Effects of siRNA Expression Vector on the Expression of Human Papillomavirus E6 Gene

目的 研究RNA干扰(RNA interference,RNAi)表达载体对宫颈癌HPV16 E6基因的抑制作用。方法 构建针对HPV16 E6基因的RNAi质粒表达载体,脂质体法转染宫颈癌caski细胞株,应用荧光定量PCR及流式细胞术检测其对HPV16 E6 m RNA及蛋白表达的影响。结果 构建的三种表达载体均抑制了HPV16 E6的m RNA及蛋白的表达,其中转染B号载体的caski B细胞HPV16 E6 m RNA表达仅相当于空白组的2 1.7% ,蛋白抑制率达98.1%。结论 RNAi表达载体可以有效的抑制HPV16 E6基因的表达。

Objective To investigate the inhibitory effects of RNAi(RNA interference)expression vector on the expression of human papilomavirus E6 gene. Methods siRNA expression vectors were constructed to be aimed directly at HPV16 E6 gene. The recombinants were transfected into cervical cancer cell line, caski, with liposomes. Expression of E6 was detected with fluorescence quantitative PCR(FQ-PCR) and flow cytometry. Results Three kinds of expression vectors could reduce the expression of E6 mRNA and protein all in caski-B cell. The expression of E6 mRNA reduced to 21.7% of control group, and the protein inhibition ratio reached 98 1%. Conclusion The RNAi expression vector can effectively inhibit the expression of HPVE6 gene.