西多福韦对HPV18阳性人宫颈癌HeLa细胞的凋亡作用及机制研究  

The apoptosis-inducing effects and the apoptosis mechanism of cidofovir for cervical cancer cell line HPV-18^+ HeLa cells in vitro

作　　者：李淑芹[1] 杨靖[1,2,3] 李蓓[2] 廖华帆 张凯[1] 杨兵[1] 李丹丹[1] 徐祥[2] 丁娜娜[3] 李明远[3] 

机构地区：[1]湖北医药学院附属人民医院肝病研究所,湖北十堰442000 [2]湖北医药学院基础医学院微生物学教研室,湖北十堰442000 [3]四川大学华西基础医学与法医学院微生物学教研室,四川成都610041

出　　处：《西部医学》2017年第5期598-603,共6页Medical Journal of West China

基　　金：2015年湖北省省级大学生创新创业训练计划项目(201510929024);湖北省教育厅科学研究计划项目(B2016113);十堰市科技局市级科学技术研究与开发专项基金项目(16K70);湖北医药学院研究生启动金资助计划项目(2015QDJZR08)

摘　　要：目的以抗肿瘤药物顺铂(Cisplatin,DDP)为阳性对照,研究抗病毒药物西多福韦(Cidofovir,CDV)对宫颈癌HPV18阳性HeLa细胞的凋亡现象和凋亡机制,探讨CDV在预防人乳头瘤病毒(HPV)感染和治疗宫颈癌方面的潜力和应用价值。方法利用流式细胞仪观察DAPI单染后HeLa细胞中凋亡细胞的形态及数目;利用Western blotting技术分析HeLa细胞中E6、p53蛋白相对表达情况;利用免疫荧光技术研究HeLa细胞中p53蛋白的亚细胞定位情况。结果流式细胞检测显示CDV和DDP能明显抑制宫颈癌HeLa细胞的增殖,使细胞周期阻滞在S期并引发细胞凋亡;Western Blotting结果显示CDV和DDP抑制HeLa细胞中E6蛋白表达,激活p53蛋白表达;细胞免疫荧光结果显示CDV和DDP可以抑制E6对p53的核输出。结论 CDV和DDP可抑制HeLa细胞增殖,呈浓度和时间依赖性;降低E6蛋白表达,恢复p53蛋白活性,进而调控细胞周期阻滞和细胞凋亡,因而CDV有可能成为宫颈癌治疗的新策略。Objective To investigate the inducement effects of Cidofovir （CDV） on apoptosis of cervical cancer cell line HeLa cells and explore the possible mechanisms, we explored the mechanism of apoptosis and offer new therapy for cervical cancer. Methods Flow Cytometry was used to detect the shape and size of apoptotic cells after DAPI staining. Western blotting was used to identify the expression of E6 and p53 protein in HeLa cells. Cell climbing immunofluores- cence technique was used to locate the subcellular position of p53 in HeLa cells. Results Flow Cytometry results showed that Cidofovir and Cisplatin both could make cells arrested in S-phase, and trigger programmed cell death （PCD）. West- ern Blotting showed that Cidofovir and Cisplatin inhibited E6 protein expression and activated p53 in HeLa cells. The immunofluorescenee results showed Cidofovir and Cisplatin could inhibit nuclear export of p53by E6 protein. Conclusion Cidofovir and Cisplatin can inhibit HeLa ceils proliferation in a concentration- and time-dependent manner, reduce the expression of E6 protein, and recover p53 protein activity. Consequently, Cidofovir regulates cell cycle arrest and apoptosis, which makes it another potential cervical cancer treatment strategy.

关 键 词：宫颈癌 人乳头瘤病毒 HELA细胞 西多福韦 顺铂 细胞凋亡 

分 类 号：R737.33[医药卫生—肿瘤]