HPV16-E6上调MIF对宫颈癌细胞增殖和凋亡的影响  

HPV16-E6 Influences Growth and Apoptosis of Cervical Cancer Cells Through Upregulation of MIF

作　　者：葛彩云 蔡红兵[1] 宋紫烨 李蓁[1] 徐梦菲 GE Caiyun;CAI Hongbing;SONG Ziye;LI Zhen;XU Mengfei(Department of Gynecological Oncology, Zhongnan Hospital of Wuhan University, Hubei Cancer Clinical Study Center, Hubei Key Laboratory of Tumor Biological Behaviors, Wuhan 430071, China)

机构地区：[1]武汉大学中南医院妇瘤科,湖北省肿瘤医学研究中心,肿瘤生物学行为湖北省重点实验室,武汉430071

出　　处：《肿瘤防治研究》2018年第7期468-474,共7页Cancer Research on Prevention and Treatment

基　　金：国家自然科学基金面上项目(81272866)

摘　　要：目的检测人乳头瘤病毒16型的致癌基因E6(HPV16-E6)通过巨噬细胞移动抑制因子(MIF)对宫颈癌细胞增殖和凋亡的影响。方法用HPV16-E6过表达和干扰质粒转染能分泌MIF的人宫颈癌细胞C33A、Si Ha和Caski。Western blot和荧光定量PCR法检测MIF蛋白和m RNA表达,并用ELISA法检测培养上清液中MIF蛋白量;将转染后的宫颈癌细胞与人巨噬细胞非接触性共培养,检测巨噬细胞中MIF表达;C33A转染HPV16-E6后加或不加MIF抑制剂,CCK-8法和流式细胞术检测细胞增殖、周期和凋亡情况;采用Pearson相关分析法分析HPV16-E6和MIF间的关系、Kaplan-Meier分析HPV16-E6对患者生存率的影响。结果癌细胞、上清液和巨噬细胞中MIF表达随HPV16-E6表达而上调(P<0.05);HPV16-E6过表达可诱导C33A细胞增殖、减少G0/G1期细胞和抑制细胞凋亡;抑制MIF后,细胞增殖率下降、凋亡率增加(P<0.05);HPV16-E6表达与MIF正相关(P<0.05),Kaplan-Meier分析显示HPV16-E6表达与患者总生存率和无进展生存率有关(P<0.05)。结论 HPV16-E6可诱导宫颈癌细胞及其微环境中巨噬细胞MIF表达,并可通过MIF诱导宫颈癌细胞增殖和抑制细胞凋亡而影响宫颈癌的预后。Objective To investigate the effect of human papillomavirus oncogene E6（HPV16-E6） on the growth and apoptosis of cervical cancer cells via macrophage migration inhibitory factor（MIF）. Methods Cervical cancer cells C33A, SiHa and Caski which could secrete MIF were transfected with HPV16-E6 overexpression vector or shRNA; Western blot, qRT-PCR and ELISA were used to examine MIF protein and mRNA expression and its supernatant. The cells above were co-cultured with human macrophages, and then we detected the expression of MIF in macrophages. MIF inhibitor was added or not in C33A group which was transfected with HPV16-E6; the cell proliferation, cell cycle and apoptosis were detected by CCK-8 method and flow cytometry. The association of HPV16-E6 expression with MIF and survival were analyzed using Pearson and Kaplan-Meier methods. Results The expression of MIF in cervical cancer cells, supernatant and macrophages were increased or decreased by up-regulation or down-regulation of HPV16-E6 expression（P〈0.05）. HPV16-E6 over-expression could induce C33A cells proliferation, shorten G0/G1 phase and inhibit cell apoptosis, and these effects were reversed using MIF inhibitor（P〈0.05）. There was positive correlation between HPV16-E6 expression and MIF in cervical cancer tissues（P〈0.05）. Survival analysis suggested the expression of HPV16-E6 in cervical cancer tissues had a significant relationship with overall survival rate and progression-free survival rate（P〈0.05）. Conclusion HPV16-E6 could induce the expression of MIF in cervical cancer cells and macrophages, promote the growth and inhibit the apoptosis of cervical cancer cells through up-regulation of MIF.

关 键 词：宫颈癌 巨噬细胞移动抑制因子(MIF) 人乳头瘤病毒16型的致癌基因E6(HPV16-E6) 凋亡 

分 类 号：R737.33[医药卫生—肿瘤]