摘要
目的:探讨α-常春藤皂苷(α-Hederin)诱导肝癌Bel-7402细胞发生铁死亡(ferroptosis)的可能机制。方法:采用梯度浓度的α-常春藤皂苷处理肝癌Bel-7402细胞24 h,MTT法检测细胞活力并计算半数抑制浓度(IC_(50))。将细胞分为空白对照组、α-常春藤皂苷(25μmol/L)组和α-常春藤皂苷(50μmol/L)组。谷胱甘肽试剂盒测定分组后细胞内GSH含量,Western blot检测铁死亡相关蛋白胱氨酸-谷氨酸的反向转运体亚基xCT、谷胱甘肽过氧化物酶4(GPX4)、二价金属转运蛋白1(DMT1)的表达,DCFH-DA探针用以检测细胞活性氧(ROS)水平。利用铁死亡抑制剂进行细胞功能恢复试验,测定了同时使用Ferrostatin-1和α-常春藤皂苷后的细胞活力、GSH含量、铁死亡相关蛋白表达水平以及ROS水平。结果:随α-常春藤皂苷浓度增加Bel-7402细胞的存活率显著降低,24 h对应Bel-7402细胞的IC_(50)为40.32μmol/L。α-常春藤皂苷能减少细胞内GSH含量,下调xCT和GPX4的表达,上调DMT1的表达和升高ROS水平。Ferrostatin-1可有效抑制α-常春藤皂苷诱导的细胞铁死亡,主要表现为细胞活力、GSH含量恢复以及ROS生成减少。结论:α-常春藤皂苷可通过抑制GPX4和xCT的表达,促进DMT1的表达,诱导ROS大量生成并导致肝癌Bel-7402细胞铁死亡。
Objective:To investigate the possible mechanism ofα-Hederin induced ferroptosis in hepatoma Bel-7402 cells.Methods:Bel-7402 cells were treated withα-Hederin at a gradient concentration for 24 h.MTT test was used to detect changes of cell viability and calculate the half-inhibitory concentration(IC_(50)).The cells were divided into blank control group,α-Hederin(25μmol/L)group andα-Hederin(50μmol/L)group.The glutathione assay kit was used to determine cellular GSH content.Western blot analysis was used to detect the expression of xCT,GPX4 and DMT1,three maker proteins in ferroptosis.DCFH-DA probe was used to detect ROS levels.Cell function recovery experiments were performed using ferroptosis inhibitor.The cell viability,GSH content,the expression levels of ferroptosis-related proteins and ROS levels were detected for the cells intervened with Ferrostatin-1 andα-Hederin.Results:The cell viability of Bel-7402 cells decreased significantly with the increase ofα-Hederin concentration,and the IC_(50) of Bel-7402 cells at 24 h was 40.32μmol/L.α-Hederin could induce a decrease of GSH content,xCT and GPX4 expression levels.On the contrary,α-Hederin could induce a increase of DMT1 expression level and ROS production.Ferrostatin-1 could effectively inhibitα-Hederin-induced ferroptosis,which was characterized by the recovery of cell viability,GSH content and reduction in ROS production.Conclusion:α-Hederin can inhibit the expression of GPX4 and xCT and promote the expression of DMT1,which leads to the massive production of ROS and ferroptosis of hepatoma Bel-7402 cells.
作者
韩松峰
关璐璐
李世朋
王梦姣
周珂欣
席守民
HAN Songfeng;GUAN Lulu;LI Shipeng;WANG Mengjiao;ZHOU Kexin;XI Shoumin(Medical Science Research Center,Basic Medical College,Henan University of Science and Technology,Henan Luoyang 471023,China;the Second Department of General Surgery,Jiaozuo People's Hospital,Henan Jiaozuo 454000,China;Henan International Joint Laboratory of Thrombosis and Hemostasis,Henan Luoyang 471023,China)
出处
《现代肿瘤医学》
CAS
北大核心
2022年第19期3467-3472,共6页
Journal of Modern Oncology
基金
河南省重点研发与推广专项(科技攻关)项目(编号:202102310126)
河南省医学科技攻关计划联合共建项目(编号:LHGJ20191342)。
作者简介
韩松峰(1994-),男,河南郑州人,硕士在读,主要从事肝癌分子生物学研究。E-mail:499354115@qq.com;通讯作者:席守民(1968-),男,河南信阳人,硕士,教授,主要从事生物化学与分子生物学研究。E-mail:xishoumin1968@163.com。
