SHC4对Erastin诱导肝癌细胞铁死亡的调控作用及机制
Regulatory effect and mechanism of SHC4 on erastin-induced ferroptosis in hepatocellular carcinoma cells
张嘉诚 1董可帅 2余佳 2王卫星1
作者信息
- 1. 武汉大学人民医院普外科 湖北 武汉 430060
- 2. 武汉大学人民医院肝胆外科 湖北 武汉 430060
- 折叠
摘要
目的:探讨Src同源2结构域转化蛋白4(SHC4)对Erastin诱导肝癌细胞铁死亡的调控作用及机制.方法:利用GEO数据库分析Erastin干扰肝癌细胞系后SHC4的变化情况.选取HepG2和Huh-7肝癌细胞系,利用慢病毒构建稳定过表达SHC4的细胞系(SHC4组)及转入空白质粒的对照组细胞系(vec组).采用CCK-8法检测过表达SHC4对铁死亡激动剂Erastin干预的肝癌活力影响,流式细胞术检测细胞活性氧(ROS)水平,丙二醛(MDA)试剂盒检测细胞MDA水平,谷胱甘肽(GSH)试剂盒检测细胞GSH水平.Western Blot检测铁死亡通路蛋白NRF2、GPX4、xCT的蛋白的表达变化.结果:Erastin干预肝癌细胞后,SHC4的表达明显增加.与对照组相比,过表达SHC4增加肝癌细胞的存活率,同时相较于vec组ROS和MDA水平降低,GSH水平升高,NRF2、xCT、GPX4蛋白明显增加.结论:SHC4通过调控NRF2-xCT/GPX4通路抑制Erastin诱导肝癌细胞铁死亡进程,进而促进肝癌的进展.
Abstract
Objective:To investigate the regulatory effect of Src homology 2 domain containing 4(SHC4)on erastin-induced ferroptosis in hepatocellular carcinoma(HCC)and the underlying mechanisms.Methods:The GEO database was used to analyze the changes in SHC4 after ferroptosis agonist eras-tin interference in liver cancer cell lines.HepG2 and Huh-7 hepatocarcinoma cell lines were selected and stably overexpressed SHC4 cell lines(SHC4 group),and blank plasmid control cell lines(vec group)were constructed by lentivirus.CCK-8 method was used to detect the effect of SHC4 overex-pression on the viability of liver cancer cells treated with Erastin.Flow cytometry was used to detect the level of reactive oxygen species(ROS),malonaldehyde(MDA),and glutathione(GSH)in cells.Western Blot was used to detect the protein expression changes of ferroptosis-related proteins,NRF2,GPX4,and xCT.Results:The expression of SHC4 was significantly increased in erastin-treated HCC cell lines.Compared with the control group,overexpression of SHC4 increased the sur-vival rate of hepatocellular carcinoma cells.Meanwhile,ROS and MDA levels decreased,GSH lev-els increased,and NRF2,xCT,and GPX4 protein expression increased significantly,compared with that respectively in the vec group.Conclusion:SHC4 inhibits ferroptosis by erastin in hepatocellular carcinoma cells by regulating the NRF2-xCT/GPX4 pathway to promote the progression of hepatocel-lular carcinoma.
关键词
SHC4/铁死亡/肝细胞肝癌/Erastin/谷胱甘肽过氧化物酶4/NRF2Key words
SHC4/Ferroptosis/Hepatocellular Carcinoma/Erastin/Glutathione Peroxidase 4/NRF2引用本文复制引用
基金项目
国家自然科学基金青年基金资助项目(82003063)
湖北省自然科学基金资助项目(2020CFB213)
出版年
2024
NETL
NSTL
万方数据