双硫仑/铜诱导肝癌细胞铁死亡的作用机制  

Mechanism for ferroptosis of hepatocellular carcinoma cells induced by disulfiram/copper

作　　者：周超婷 敬强安 张萍[2] 吴韵怡 任雪营 王莹 饶国武[1] 童向民[2] ZHOU Chaoting;JING Qiang′an;ZHANG Ping(College of Pharmacy,Zhejiang University of Technology,Hangzhou310014,CHINA)

机构地区：[1]浙江工业大学药学院,浙江杭州310014 [2]浙江省人民医院临床医学研究所 [3]浙江省新华医院医学检验科 [4]杭州市第一人民医院GCP机构办公室

出　　处：《江苏医药》2023年第4期330-333,F0003,共5页Jiangsu Medical Journal

基　　金：浙江省医药卫生科技计划(2021KY842)。

摘　　要：目的 探讨双硫仑/铜(DSF/Cu)诱导肝癌细胞铁死亡的作用机制。方法 采用DSF/Cu 0、0.5、1.0、1.5、2.0和4.0μmol/L处理人肝癌细胞株Huh7,CCK-8法检测细胞活力,激光共聚焦显微镜观察细胞内脂质过氧化物和谷胱甘肽(GSH)水平的变化,流式细胞术检测细胞内活性氧(ROS)水平,Western blot和实时荧光定量PCR检测铁死亡相关基因谷胱甘肽过氧化物酶4(GPX4)的蛋白和mRNA表达。结果 随着DSF/Cu浓度的升高,细胞活力逐渐降低(P<0.05)。与DSF/Cu 0μmol/L比较,DSF/Cu处理细胞后,脂质过氧化物积累,细胞内ROS水平升高。与DSF/Cu 0μmol/L比较,DSF/Cu 2.0μmol/L能够下调细胞GPX4蛋白和mRNA的表达(P<0.05),降低细胞GSH水平(P<0.05)。铁死亡抑制剂去铁胺能够部分恢复DSF/Cu抑制的细胞活力(P<0.05)。结论 DSF/Cu可抑制肝癌细胞活力,并通过抑制GPX4表达来诱导肝癌细胞发生铁死亡。Objective To explore the mechanism for ferroptosis of hepatocellular carcinoma cells induced by disulfiram/copper(DSF/Cu).Methods Human hepatocellular carcinoma cell line Huh7was treated with DSF/Cu 0,0.5,1.0,1.5,2.0and 4.0μmol/L.The cell viability was detected by CCK-8method.The changes of lipid peroxide and glutathione(GSH)levels in the cells were observed by laser scanning confocal microscope.The intracellular reactive oxygen species(ROS)levels were detected by flow cytometry.The protein and mRNA expressions of glutathione peroxidase 4(GPX4)were detected by Western blot and real-time quantitative PCR,respectively.Results With the increase of DSF/Cu concentration,the cell viability was decreased gradually(P<0.05).Compared with 0μmol/L,the lipid peroxide was accumulated and intracellular ROS level was increased in the cells treated with DSF/Cu.Compared with 0μmol/L,DSF/Cu 2.0μmol/L downregulated the expressions of GPX4protein and mRNA and reduced the level of GSH(P<0.05).The ferroptosis inhibitor DFO could partially restore the cell viability inhibited by DSF/Cu(P<0.05).Conclusion DSF/Cu can inhibit the cell viability of the hepatocellular carcinoma cells and induce the ferroptosis by inhibiting GPX4.

关 键 词：双硫仑 铜 谷胱甘肽过氧化物酶4 肝细胞癌 铁死亡 

分 类 号：R735[医药卫生—肿瘤]