人乳头瘤病毒16E7和CulL2依赖蛋白对APOBEC3A蛋白表达及子宫颈细胞迁徙能力的影响  

Effects of Human Papillomavirus 16 E7 and CulL2-dependent Proteins on APOBEC3A Protein Expression and Cervical Cell Migration

作　　者：程娟[1] 卢永丽 杨文君[3] CHENG Juan;LU Yongli;YANG Wenjun(Surgical System Teaching and Research Section,Department of Medicine,Ya'an Vocational College,Ya'an 625000,China;Department of Obstetrics and Gynecology,Affiliated Hospital of Ya'an Vocational College,Ya'an 625000,China;Biochemistry Teaching and Research Group,Department of Pharmaceutical Sciences,Ya'an Vocational College,Ya'an 625000,China)

机构地区：[1]雅安职业技术学院医学系外科系统教研室,雅安625000 [2]雅安职业技术学院附属医院,妇产科雅安625000 [3]雅安职业技术学院药学检验系生物化学教研组,雅安625000

出　　处：《病毒学报》2019年第3期402-407,共6页Chinese Journal of Virology

摘　　要：人乳头瘤病毒(Human papillomavirus,HPV)16 E7基因编码多功能磷酸蛋白,其功能和结构均类似腺病毒,HPV 16 E7可与激活的致癌基因结合,促进肿瘤进展。但是HPV 16 E7对子宫颈细胞迁移的影响和机制尚不清楚。因此本研究探究了HPV16 E7和CulL2依赖蛋白对APOBEC3A蛋白表达及子宫颈细胞迁徙能力的影响。随机选择60名年龄在21～55岁间的妇女,分别设置空白对照组(B组)、患有人乳头瘤细胞组(P组)、添加CulL2依赖蛋白组(C组),采用流式荧光杂交法和第二代杂交捕获实验(hc2)检测其子宫颈脱落细胞,进行细胞培养,采用Transwell法检测细胞数,重组质粒DNA分析基因克隆的完整性,代谢标记细胞,进行过氧化氢酶(catalase,CAT)分析。采用实时荧光定量PCR法对CulL2和APOBEC3A基因水平进行评估测定,免疫印迹(Western Blot,WB)检测相关蛋白表达。与B组相比,P组细胞穿透基质胶的细胞数增多,即迁徙能力提高,重组质粒转染后,C组细胞穿透基质胶的细胞数亦明显增多,但增多幅度不如P组,即迁徙能力有所增强(P<0.05)。高风险HPV蛋白E7可上调细胞中APOBEC3A蛋白的表达。APOBEC3A和HPV16 E7与CulL2相互作用,表明在HPV感染期间形成的E7可调节细胞中的APOBEC3A蛋白的水平。反转录PCR(Reverse transcription-PCR,RT-PCR)结果显示,与B组比较,P组细胞基因表达增加,C组细胞中CulL2和APOBEC3A基因的mRNA表达水平明显减少(P<0.05)。本研究发现HPV16 E7和CulL2依赖蛋白可影响APOBEC3A蛋白表达,并且促进子宫颈细胞的迁徙能力。The human papillomavirus(HPV)16 E7 gene encodes a multifunctional phosphoprotein with func-tions and structures similar to those of adenovirus.HPV 16 E7 binds to activated oncogenes and promotes tumor progression.However,the effect and mechanism of HPV 16 E7 on cervical cell migration remains unclear.So,we want to investigate the effects of HPV 16 E7 and CulL2-dependent proteins on the expression of APO-BEC3A protein and the migration ability of cervical cells in this study.Sixty women aged 21~55 years were ran-domly selected,and a blank control group(Group B),a human papilloma cell group(Group P),and a CulL2-dependent protein group(Group C)were added.Flow cytometry hybridization and second-generation hybrid capture assay(hc2)were used to detect cervical exfoliated cells in cell culture.Cell number was detected by Transwell method.Recombinant plasmid DNA was used to analyze the integrity of gene clones,and metaboli-cally labelled cells were used for CAT analysis.The levels of CulL2 and APOBEC3A genes were evaluated by real-time fluorescent quantitative PCR,and the expression of related proteins was detected by Western Blot.Compared to Group B,the number of cells penetrating matrix gelatin in Group P was increased,indicating the migration ability was improved.After transfection with recombinant plasmid,the number of cells penetrating matrix gelatin in Group C also increased significantly,but less than that in Group P,suggesting the migration ability was enhanced(P<0.05).HPV protein E7 can up-regulate the expression of APOBEC3A in cells.APO-BEC3A and HPV16 E7 interact with CulL2,indicating levels of APOBEC3A protein in E7 regulatable cells were formed during HPV infection.RT-PCR results showed that compared to Group B,the gene expression in Group P was increased,and the mRNA expression levels of CulL2 and APOBEC3A genes in Group C were significantly decreased(P<0.05).HPV 16 E7 and CulL2-dependent proteins can affect the expression of APO-BEC3A protein and promote the migration of cervical cells.

关 键 词：人乳头瘤病毒(HPV) CulL2依赖蛋白 APOBEC3A蛋白 蛋白降解 

分 类 号：R373.9[医药卫生—病原生物学] R511[医药卫生—基础医学]