HPV16 E6对高迁移率族蛋白B1主动释放及宫颈癌细胞侵袭能力的影响  

The effects of HPV16 E6 on active release of high mobility group box-B1 and the invasion ability of cervical cancer cells

作　　者：王景[1] 李伟[2] 蔡红兵[3] 程欣[3] 

机构地区：[1]襄阳市中心医院(湖北文理学院附属医院)妇产科,湖北襄阳441021 [2]襄阳市第一人民医院(湖北医药学院附属襄阳医院)妇产科,湖北襄阳441000 [3]武汉大学中南医院妇瘤科,湖北省肿瘤医学临床研究中心,肿瘤生物学行为湖北省重点实验室,湖北武汉430071

出　　处：《肿瘤》2013年第4期309-313,共5页Tumor

基　　金：国家自然科学基金面上项目(编号:81272866)

摘　　要：目的:探讨人乳头瘤病毒(human papillomavirus,HPV)16E6对宫颈癌细胞中高迁移率族蛋白B1(high mobility group box-B1,HMGB1)主动释放及细胞侵袭能力的影响。方法:构建HPV16E6表达质粒,并转染至宫颈癌C33A细胞,FCM法分析细胞存活率;半定量RT-PCR法检测转染24h前后C33A细胞中HPV16E6基因的表达水平;蛋白质印迹法检测转染前后C33A细胞的细胞核、细胞质以及细胞培养液中HMGB1蛋白水平的变化情况;Transwell实验检测细胞侵袭能力的变化。结果:HPV16E6基因转染C33A细胞后,细胞核中HMGB1蛋白水平明显降低(P<0.01),而细胞质及细胞培养液中HMGB1蛋白水平均明显升高(P<0.01)。转染后细胞存活率未见明显差异,但细胞穿膜个数明显多于未转染组(P<0.01)。结论:HPV16E6可诱导宫颈癌细胞内HMGB1的主动释放,提高肿瘤细胞的侵袭能力。Objective： To explore the effects of HPV （human papillomavirus）16 oncogene E6 on active release of HMGB1 （high mobility group box-B1） and invasive ability of cervical cancer cells. Methods： HPV16 E6 recombinant plasmid was constructed and verified by restriction enzyme digestion and DNA sequencing. The survival rate of C33A cells which were transfected with HPV16 E6 recombinant plasmids was analyzed by FCM （flow cytometry）. The expression of HPVl 6 E6 gene was detected by semi- quantitative RT-PCR （reverse transcription-PCR） in C33A cells before （as the control group） and after transfection with HPV16 E6 recombinant plasmids for 24 h, then the level changes of HMGB1 protein in nucleus, cytoplasms and culture medium detected by Western blotting in different groups. The change of invasive ability of C33A cells after transfection was detected by Transwell method. Results： The expression level of HMGB1 protein in nucleus of C33A cells in the transfection group was lower than that in the control group （P 〈 0.01）, while the expression of HMGB1 protein significantly increased in cytoplasm and cell medium （P 〈 0.01）. The number of C33A cells go through the membrane in the transfection group was significantly more than that in the control groups （P 〈 0.01）, but there was no difference in survival rate of C33A cells between the two groups. Conclusion： HPV16 E6 can facilitate the active release of HMGB1 in cervical cancer cells, and then enhance their abilities of invasion and migration.

关 键 词：宫颈肿瘤 高迁移率族蛋白质类 人乳头瘤病毒16 肿瘤浸润 

分 类 号：R737.33[医药卫生—肿瘤]