HPV16E6肺癌细胞模型的构建

Establishment of HPV16E6 lung cancer cell model

目的构建HPV16E6肺癌细胞模型，为研究针对HPV16E6特异性抗原的肿瘤免疫奠定基础。方法①人工合成HPV16E6基因序列，构建HPV16E6基因的重组质粒载体；②将慢病毒包被含HPV16E6基因的pEGFP—N1质粒载体并感染H1299肺癌细胞，经RT-PCR和Westernblot检测，验证HPV16E6基因在H1299肺癌细胞的过表达；③构建HPV16E6肺癌细胞裸鼠模型，免疫组化验证HPV16E6蛋白在肺癌细胞中的表达。结果①成功构建含HPV16E6基因的重组质粒载体；②成功将慢病毒感染H1299肺癌细胞，经实时定量PCR和Western blot检测成功验证HPV16E6基因在H1299肺癌细胞中过表达；③成功构建HPV16E6肺癌细胞裸鼠模型，成功验证HPV16E6蛋白在肺癌细胞中的表达。结论使用慢病毒感染方法成功构建了稳定的HPV16E6肺癌细胞模型。

Objective To construct HPV16E6 lung cancer cell models to lay the foundation for tumor immunity research which was directed to HPV16E6 specific antigen. Methods （1）HPV16E6 gene sequences were artificially synthesized and recombinant plasmid vectors containing HPV16E6 gene were constructed.（2） pEGFP-N1 plasmid vectors containing HPV16E6 gene were packaged by lentivirus and infected H1299 lung cancer cells. HPV16E6 gene overexpression in H1299 lung cancer cells was verified by RT-PCR and Western blot. （3）Nude mice models containing lung cancer cells were contructed and HPV16E6 protein expression in lung cancer cells was verified by immunohistochernistry. Results （1） Recombinant plasmid vectors containing HPV16E6 gene were constructed successfully. （2）H1299 lung cancer cells were successfully infected by lentivirus and HPV16E6 gene overexpression in H1299 lung cancer cells was successfully verified by RT-PCR and Western blot. （3）Nude mice models containing lung cancer cells were successfully contruscted and HPV16E6 protein expression in lung cancer cells was successfully verified by immunohistochemistry. Conclusions Stable model containing lung cancer cells could be successfully contruscted by lentivirus infection.