中文摘要：

目的 研究子宫颈癌C33A细胞中人乳头瘤病毒16 E6（HPV16 E6）蛋白过表达对内质网应激-自噬通路的影响.方法 PCR检测子宫颈癌C33A细胞中HPV的整合情况;构建HPV16 E6的真核表达载体pcDNA3.1--HPV16 E6,转染C33A细胞,即pcDNA3.1--HPV16 E6组,同时设空载体转染组pcDNA3.1-组和空白对照组（C33A组）,Western blot检测C33A细胞中HPV 16 E6蛋白表达的变化及对自噬基因Beclin 1、LC3Ⅱ、内质网应激相关基因IRE1、PERK和ATF6蛋白表达的影响.结果 C33A细胞中无HPV整合,选为干预细胞株;成功构建了HPV16 E6的真核表达载体pcDNA3.1--HPV16 E6,其可以使C33A细胞中HPV16 E6蛋白表达增强,HPV16 E6过表达可使Beclin 1、LC3Ⅱ和IRE1、PERK、ATF6蛋白表达增加,与pcDNA3.1-组和C33A组相比,差异有统计学意义（P＜0.05）.阻断内质网应激通路后,LC3Ⅱ蛋白表达降低.结论 HPV16 E6蛋白能促使子宫颈癌C33A细胞自噬增强,该过程是通过内质网应激途径实现的,这一反应可能参与HPV16致子宫颈癌的过程,为子宫颈癌的防治提供了新思路.

英文摘要：

Objective To investigate the effects of human papilloma virus 16 E6 （HPV16 E6） on endoplasmic reticulum （ER） stress-autophagic response in the cervical cancer C33A cells.Methods Polymerase chain reaction was used for detecting the integration of HPV DNA.The eukaryotic expression vector of HPV16 E6 was constructed and transfected via lipofectamine into C33A cells.Experimental cells were classified into 3 groups：pcDNA3.1--HPV16 E6 group,pcDNA 3.1-group and C33A group.Western blot was used to measure expression of protein of HPV16 E6,Beclin 1,LC3 Ⅱ,IRE1,PERK and ATF6 in transfected cells.Results here was no HPV DNA integration in C33A cells that were confirmed as the intervention cells.Eukaryotic expression vector pcDNA3.1--HPV16 E6 was constructed successfully.The eukaryotic expression vector pcDNA3.1--HPV16 E6 significantly improved the expression of protein of HPV 16 E6 in C33A cells.The protein expression of Beclin 1,LC3 Ⅱ,IRE1,PERK and ATF6 were significantly improved after transfection with vector pcDNA3.1 ＋-HPV16 E6 （P 〈 0.05）.Furthermore,LC3 Ⅱ protein level was reduced by treatment with ER stress inhibitor.Conclusion HPV16 E6 can improve autophagy through the ER stress pathway,and this response may play an important role in the process of HPV16 E6 inducing cervical cancer,providing one of the new strategies for gene therapy of cervical carcinoma.