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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 study the inhibitory effect of RNA interference (RNAi) expression vector on HPV16 E6 gene in cervical cancer. Methods RNAi expression vector targeting HPV16 E6 gene was constructed and transfected into cervical cancer cell line caski by lipofectamine. Fluorescent quantitative PCR and flow cytometry were used to detect the expression of HPV16 E6 mRNA and protein. Results The expression vectors of m RNA and protein of HPV16 E6 were all inhibited by the three expression vectors. The expression of HPV16 E6 mRNA in the caski B cell transfected with B vector was only 21.7% of the blank group, and the protein inhibition rate was 98.1 %. Conclusion RNAi expression vector can effectively inhibit the expression of HPV16 E6 gene.