索拉菲尼诱导的细胞凋亡和自噬对HeLa细胞耐药性的影响  

Effect of sorafenib induced apoptosis and autophagy on drug resistance in HeLa cells

作　　者：杨铠菲 朱静格 张洋洋 赵俊果 高雨悦 胡焕焕 姬国杰 YANG Kaifei;ZHU Jingge;ZHANG Yangyang;ZHAO Junguo;GAO Yuyue;HU Huanhuan;JI Guojie(Department of School of Life Science and Technology,Sanquan College of Xinxiang Medical University,Xinxiang 453000,China;Key Laboratory of Fertility,Sanquan College of Xinxiang Medical University,Xinxiang 453000,China;Experimental Teaching Center of Biologyand Basic Medical Sciences,Sanquan College of Xinxiang Medical University,Xinxiang 453000,China)

机构地区：[1]新乡医学院三全学院生命科学技术学院,河南新乡453000 [2]新乡医学院三全学院生育力保存重点实验室,河南新乡453000 [3]新乡医学院三全学院生物与基础医学实验教学中心,河南新乡453000

出　　处：《基础医学与临床》2024年第4期467-473,共7页Basic and Clinical Medicine

基　　金：河南省科技攻关项目(232102310303);新乡医学院三全学院骨干教师培养计划(SQ2023GGJS06);新乡医学院三全学院学术技术带头人培养计划(SQ2023XSJSDTR01)。

摘　　要：目的 探究索拉菲尼(sorafenib)通过细胞凋亡和自噬对人宫颈癌细胞系HeLa增殖的影响,及对HeLa细胞耐药性的影响。方法 用间歇诱导法构建耐药细胞株,以0、2.5、5.0、7.5、10.0、15.0、20.0μmol/L为梯度的索拉菲尼处理HeLa细胞,每个浓度维持1周,并筛选出可稳定传代的耐药细胞株;MTT法检测索拉菲尼对细胞增殖的影响;流式细胞术分析细胞周期分布情况;RT-qPCR检测亲本株和耐药株在不同药物浓度下耐药和凋亡基因表达水平的变化。Western blot检测与自噬相关的标志物蛋白LC3-Ⅰ和LC3-Ⅱ的变化。结果 成功获得稳定的耐药株;经药物处理的细胞更多的被阻滞在G1期。在耐药细胞中,凋亡抑制基因Bcl-2表达量显著降低;凋亡基因Bax表达量明显增加;耐药基因表达明显升高。Western blot中耐药细胞LC3-Ⅱ/LC3-Ⅰ显著高于亲本细胞(P<0.05)。结论 索拉菲尼通过阻滞细胞周期抑制HeLa细胞增殖并诱导其自噬。细胞自噬在宫颈癌治疗过程中可能参与了细胞耐药的发展,促进细胞存活;在一定程度下,药物诱导的细胞自噬能激活耐药细胞的凋亡信号通路,促进细胞耐药逆转。Objective To explore the effect of sorafenib on HeLa cell proliferation by inducing cell apoptosis and autophagy and its impact on drug resistance.Methods The drug-resistant cell strains were constructed through intermittent induction method,with concentrations of 0,2.5,5.0,7.5,10.0,15.0,20.0μmol/L.HeLa cells were incubated with increasing concentrations of sorafenib with each concentration for 1 week.The drug-resistant cell strains with stable passages were collected.MTT assay was used to detect the effect of sorafenib on cell proliferation.Cell cycle distribution was analyzed by flow cytometry.The change in the expression of drug-resistant and apoptotic genes in the parents and drug-resistant cell strains under different drug concentrations was examined by semi-quantitative PCR.The changes of apoptotic related marker proteins LC3-Ⅰand LC3-Ⅱwere detected by Western blot.Results Stable drug-resistant strains were successfully obtained;Drug-treated cells were more blocked in the G1 phase.In drug-resistant cells,the expression of apoptosis suppressor gene Bcl-2 was significantly decreased and the apoptotic gene Bax as well as the drug-resistant genes were all significantly increased(P<0.05).The LC3-Ⅱ/LC3-Ⅰratio of drug-resistant cells was significantly higher than that of parent cells(P<0.05).Conclusions Sorafenib may block the cell cycle,suppress malignant cell proliferation and promote autophage.On one hand,autophagy participates in the development of cell drug resistance and promotes cell survival.On the other hand,drug-induced autophagy may activate some of apoptotic signaling pathway in drug-resistant cells and promote the reversal of cell drug resistance.

关 键 词：索拉菲尼 宫颈癌 细胞耐药性 自噬 凋亡 细胞周期 

分 类 号：R737.33[医药卫生—肿瘤]