【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。更多还原

【Abstract】 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 p53protein in HeLa cells.Cell climbing immunofluorescence 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).Western Blotting showed that Cidofovir and Cisplatin inhibited E6 protein expression and activated p53 in HeLa cells.The immunofluorescence results showed Cidofovir and Cisplatin could inhibit nuclear export of p53 by E6protein.Conclusion Cidofovir and Cisplatin can inhibit HeLa cells 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.更多还原