AIFM2抑制铁死亡介导肝癌细胞索拉菲尼耐药  

AIFM2 mediates sorafenib resistance by inhibiting ferroptosis in hepatocellular carcinoma cells

作　　者：刘帆[1] 魏士博[1] LIU Fan;WEI Shi-bo(Department of General Surgery,the Fourth Affiliated Hospital of China Medical University,Shenyang 110032,China)

机构地区：[1]中国医科大学附属第四医院普外科,辽宁沈阳110032

出　　处：《解剖科学进展》2022年第5期567-572,共6页Progress of Anatomical Sciences

基　　金：辽宁省高等学校科学研究经费项目(LJKZ0770)。

摘　　要：目的探讨AIFM2对肝癌细胞索拉菲尼耐药的影响及对铁死亡的调控。方法采用实时定量PCR检测30例肝细胞癌组织与相应癌旁组织和肝癌细胞系及正常肝细胞系中AIFM2 mRNA的表达水平;采用浓度梯度法培养索拉非尼耐药肝癌细胞,sh-AIFM2转染肝细胞癌细胞以抑制AIFM2表达,sh-Ctrl作为对照,实时定量PCR检测转染效率,Western blot法检测转染后AIFM2蛋白的表达;CCK-8法检测细胞活力,克隆形成实验检测细胞的增殖能力;DCFH-DA染色法检测ROS表达,脂质过氧化试剂盒检测MDA的表达,微量还原型谷胱甘肽(GSH)试剂盒检测MDA表达,流式细胞术检测铁离子的表达。结果AIFM2在肝细胞癌组织中相对癌旁高表达,AIFM2在肝癌细胞中的表达高于正常肝细胞系;AIFM2基因敲低促进肝癌耐药细胞活性的下降,并促进肝癌耐药细胞发生铁死亡,其中ROS、MDA表达增加,GSH表达降低,而铁离子表达并没有发生明显改变。结论AIFM2抑制铁死亡介导肝癌细胞索拉非尼耐药,联合应用AIFM2抑制剂和索拉非尼可能是治疗肝细胞癌耐药的有效途径。Objective To investigate the effect of AIFM2 on sorafenib resistance and ferroptosis in hepatocellular carcinoma cells.Methods Real time quantitative PCR was used to detect the expression of AIFM2 mRNA in 30 cases of hepatocellular carcinoma and corresponding adjacent tissues,and to detect the expression of AIFM2 mRNA in hepatocellular carcinoma cell lines and normal liver cell lines.Sh-AIFM2 was transfected into HCC cells to inhibit the expression of AIFM2.The transfection efficiency was detected by real-time quantitative PCR,and the expression of AIFM2protein was detected by Western blot.CCK-8 assay was used to detect cell viability,and clone formation experiment was used to detect cell proliferation.The expression of ROS was detected by DCFH-DA staining.The expression of MDA was detected by lipid peroxidation kit.The expression of GSH was detected by micro reduced glutathione(GSH)kit,and the expression of iron ions was detected by flow cytometry.Results The expression of AIFM2 in HCC tissues was higher than that in adjacent normal tissues,and the expression of AIFM2 in HCC cells was higher than that in normal liver cell line.Knockdown of AIFM2 decreased the cell viability of sorafenib resistance cells and promoted ferroptosis.Knockdown of AIFM2 promoted the level of ROS,MDA and inhibited the expression of GSH,however,the expression of iron ions did not change significantly.Conclusion AIFM2 inhibits ferroptosis and mediates sorafenib resistance in hepatocellular carcinoma cells.The combination of AIFM2 inhibitors and sorafenib may be an effective way to treat hepatocellular carcinoma drug resistance.

关 键 词：肝细胞癌 索拉菲尼耐药 铁死亡 AIFM2 

分 类 号：R735.7[医药卫生—肿瘤]