基于HPV16的新型伪病毒肿瘤疫苗对Hela细胞的杀伤作用研究

Study on killing effect of pseudovirus tumor vaccine of human papilloma virus type 16 on Hela cells

目的:构建基于人乳头状瘤病毒(human papilloma virus,HPV)16型的肿瘤疫苗,并检测其对Hela肿瘤细胞的杀伤活性。方法:利用昆虫杆状病毒表达系统表达病毒蛋白,在体外变性与复性过程中将病毒蛋白包装绿色荧蛋白EGFP和白喉毒素(Diphtheriatoxin,DT)A链(DT-A)的双表达质粒,形成伪病毒疫苗。透射电镜观察病毒样颗粒(virus-like particle,VLP)的结构,并转染Hela肿瘤细胞,荧光显微镜和流式细胞仪检测其转染效率和对Hela肿瘤细胞的杀伤能力。结果:透射电镜观察证实病毒蛋白可自我组装成VLP,在转染Hela肿瘤细胞后,荧光显微镜和流式细胞仪成功检测到荧光的表达和对Hela肿瘤细胞的杀伤作用。结论:基于HPV16的新型伪病毒肿瘤疫苗能成功转染Hela肿瘤细胞并产生杀伤活性,为肿瘤的基因治疗提供了依据。

Objective：To construct human papilloma virus type 16 （HPV16） tumor vaccine and to examine its kill activity on Hela cells. Methods： Virus protein was expressed by Bac -to -Bac expression system, EGFP and DT - A double expression plasmid together with virus protein were formed into pseudovirus in the course of degenera- tion and regeneration in vitro. The construction was identified by transmission electron microscope, the transfection and kill activity were detected by fluoroscope and FCM. Results： The pseudovirus could selfassemble into VLP, fluoroscope and FCM could detect the expression of fluorescence and kill activity successfully. Conclusion： Pseud- ovirus tumor vaccine of human papilloma virus type 16 could transfect and kill Hela cells, which might provide a new strategy for tumor gene therapy.