慢病毒介导的HPV16-E7基因RNAi细胞模型的建立

 建立HPV16-E7基因的RNAi细胞模型，可为进一步研究HPV16-E7蛋白作用及致癌机制提供物质前提。选择HPV16-E7基因特异的siRNA片段，构建慢病毒siRNA重组表达载体，经293FT病毒包装细胞转染产生重组病毒，并以此感染HPV16阳性SiHa宫颈癌细胞、抗菌素筛选和分子生物学鉴定，建立了稳定表达HPV16-E7-siRNA的RNAi细胞模型。该细胞模型，对目标基因（E7）表达的抑制效率，也以蛋白免疫印迹和RT-PCR确证。由此得出，利用慢病毒载体和HPV16-E7基因特异的siRNA片段，可以建立一种稳定、有效和可行的RNAi细胞模型，为进一步研究E7蛋白的作用及致癌机制奠定了重要的物质基础。

Construction of Lentivirus-mediated HPV16-E7 Gene RNAi Cell Model

The HPV16-E7 ggene RNAi cell model was constructed. A recombinant lentiviral siRNA expression vector was constructed using a HPV16-E7 oncogene specific siRNA fragment, and an RNAi cell model stably expressing the HPV16-E7-siRNA was established by transfection of 293FT virus-packiging cells with the viral vector followed by infection of HPV16-positive SiHa carcinoma cells with recombinant virus, antibiotica selection and molecular biological characterization. RNAi cell model was successfully established, stably transcribing the HPV16-E7-siRNA fragment, and the inhitory effect on target gene (E7) was also characterized by western blotting and RT-PCR. The RNAi cell model expressing the HPV16-E7 gene-specific siRNA fragment using lentiviral vector was stable, effective and feasible, which lays the foundation for the role study and carcinogenic mechanism of E7 protein.