HPV16 E6对ZEB1介导的宫颈癌细胞上皮间质转化的促进作用  

Promoting Effect of HPV16 E6 on ZEB1-Mediated EMT in Cervical Cancer Cells

作　　者：赵文君 左娜 葛彩云 蔡红兵[1] ZHAO Wenjun;ZUO Na;GE Caiyun;CAI Hongbing(Dept.of Gynecological Oncology,Zhongnan Hospital of Wuhan Universit;Hubei Key Laberatory of Tumor Biological Behavir;Hubei Cancer Clinical Study Center,Wuhan 430071,China)

机构地区：[1]武汉大学中南医院妇瘤科/肿瘤生物学行为湖北省重点实验室/湖北省肿瘤医学临床研究中心

出　　处：《武汉大学学报（医学版）》2018年第5期693-698,共6页Medical Journal of Wuhan University

基　　金：国家自然科学基金资助项目(编号:81272866)

摘　　要：目的：探讨宫颈癌细胞中HPV16E6病毒蛋白对E盒结合锌指蛋白1（ZEB1）及其介导的上皮间质转化（EMT）的影响。方法：构建HPV16E6质粒并转染至宫颈癌细胞C33A（E6-C33A组）,同时设置Control-C33A组;设计合成靶向E6特异性shRNA,使Siha细胞中E6低表达（shE6-Siha组）,设置Control-Siha组。划痕实验比较各组细胞迁移情况;荧光定量PCR和Western Blot检测EMT相关指标（E-cadherin、vimentin）及ZEB1的表达;生物信息学方法预测HPV16E6和ZEB1的三级结构及二者的相互作用,免疫共沉淀进行验证。结果：划痕实验显示E6-C33A组细胞迁移能力显著高于Control-C33A组;而shE6-Siha组细胞迁移能力显著低于Control-Siha组;与Control-C33A组相比,E6-C33A组中ZEB1、vimentin的表达量明显升高,E-cadherin表达则降低;与Control-Siha组相比,shE6-Siha组ZEB1、vimentin表达下降,E-cadherin表达升高。免疫共沉淀证明HPV16E6与ZEB1存在相互作用。结论：HPV16E6可促进ZEB1的表达,诱发宫颈癌细胞的EMT。Objective：To explore the influence of HPV16 E6 on the expression of zinc finger E-box binding homobox1（ZEB1）and the ZEB1-mediated Epithelial-to-mesenchymal transition（EMT）in cervical cancer cells.Methods：In this study,recombinant plasmid HPV16 E6 was transfected into cervical cancer cells C33 Ato gain a cell line which can express HPV16 E6 while the empty vector was used as the control.The target-specific shRNA of E6 was well designed according to its gene sequence and transfected into cervical cancer cell SiHa to gain a cell line in which the expression level of E6 can be lowered while the empty vector was used as the control.The expression level of EMT related indicators（E-cadherin,vimentin）and ZEB1 was measured by quantitative PCR and Western Blot.Bioinformatics methods were used to predict the tertiary structure of HPV16 E6 and ZEB1 and interaction between them,and the results were confirmed by Co-immunoprecipitation.Results：The scratch test showed that the migration ability of E6-C33 Agroup was significantly higher than the control-C33 Agroup,while the cell migration ability of shE6-Siha group was significantly lower than the control-Siha group.Compared with the control-C33 Agroup,the expressions of ZEB1 and vimentin were much higher in E6-C33 Agroup,while the expression of E-cadherin was lower notably.Compared with control-Siha,vimentin and ZEB1 expression decreased,and E-cadherin expression increased in shE6-Siha.Co-immunoprecipitation showed that HPV16 E6 interacts with ZEB1.Conclusion：HPV16 E6 can promote the expression of ZEB1 and accelerate ZEB1-mediated EMT in cervical cancer cells.

关 键 词：宫颈癌 HPV16 E6 E盒结合锌指蛋白1 上皮-间质转化 

分 类 号：R737.6[医药卫生—肿瘤]