华蟾素通过调控HIF1α的SUMO修饰促进胃癌细胞铁死亡  

Cinobufotalin promotes ferroptosis of gastric cancer cells by regulating the SUMO modification of HIF1α

作　　者：郑梓莹 张燕 陈文[1] 林玉婷 叶磊[1] ZHENG Zi-ying;ZHANG Yan;CHEN Wen;LIN Yu-ting;YE Lei(Dept of Traditional Chinese Medicine,The Second Attached Hospital of Fujian Medical University,Quanzhou Fujian 362000,China)

机构地区：[1]福建医科大学附属第二医院中医科,福建泉州362000

出　　处：《中国药理学通报》2024年第7期1342-1349,共8页Chinese Pharmacological Bulletin

基　　金：福建省自然科学基金项目(No 2021J01270)。

摘　　要：目的本研究旨在探讨华蟾素(cinobufotalin,CB)调控缺氧诱导因子1α(hypoxia inducible factor 1 alpha,HIF1α)的小泛素样修饰相关蛋白(small ubiquitin-like modifier,SUMO)修饰对胃癌细胞铁死亡的影响。方法使用不同浓度CB处理人正常胃黏膜上皮细胞(GES-1)和人胃癌细胞(MGC803)。MTT检测细胞活力并计算CB在胃癌细胞中的IC_(50)值,Transwell检测侵袭。铁死亡激动剂(erastin)或抑制剂(ferrostatin-1)处理癌细胞并评估胃癌细胞脂质活性氧(reactive oxygen species,ROS)水平、MDA水平和细胞凋亡、细胞内总铁水平和Fe^(2+)的水平。Western blot检测SUMO1和HIF1α的表达,免疫沉淀(immunoprecipitation,IP)检测SUMO1和HIF1α的相互作用。建立异种移植瘤模型,使用CB或erastin治疗,评估CB在体内对肿瘤生长和胃癌细胞铁死亡的影响。结果2μmol·L^(-1)以下CB未对GES-1细胞活力产生影响。与Con组相比,CB处理剂量依赖性地抑制MGC803细胞活力和侵袭。与Con组比较,CB处理提高胃癌细胞脂质ROS、MDA、总铁和Fe^(2+)水平,并促进细胞凋亡(均P<0.05)。CB联合erastin增强铁死亡,而ferrostatin-1处理则抑制CB诱导的胃癌细胞铁死亡。机制上,CB抑制SUMO1表达,减少HIF1α的SUMO化修饰,进而抑制其表达。CB诱导的胃癌细胞铁死亡能够被SUMO1过表达载体逆转。体内实验证实,CB能够抑制肿瘤生长并诱导胃癌细胞铁死亡。结论CB通过抑制HIF1α的SUMO化修饰进而诱导胃癌细胞铁死亡。Aim To investigate the effect of cinobufotalin(CB)on the small ubiquitin-like modifier(SUMO)modification of hypoxia inducible factor 1 alpha(HIF1α)on ferroptosis of gastric cancer cells.Methods Human normal gastric mucosal epithelial cells(GES-1)and gastric cancer cells(MGC803)were treated with various concentrations of CB.MTT assay was employed to determine cell viability and calculate the IC 50 value of CB in gastric cancer cells.Cell invasion was evaluated using Transwell assay.Cancer cells were subjected to treatment with ferroptosis agonists(erastin)or inhibitors(ferrostatin-1)to assess the levels of lipid reactive oxygen species(ROS),malondialdehyde(MDA),cell apoptosis,intracellular total iron,and Fe^(2+)in gastric cancer cells.Western blot analysis was conducted to detect the expression of SUMO1 and HIF1α,while immunoprecipitation(IP)was utilized to investigate the interaction between SUMO1 and HIF1α.An allograft tumor model was established and treated with CB or erastin to assess the impact of CB on tumor growth and ferroptosis in gastric cancer cells in vivo.Results CB at concentrations below 2μmol·L^(-1)had no impact on the viability of GES-1 cells.Compared to the control group,CB treatment dose-dependently inhibited the viability and invasion of MGC803 cells.CB treatment increased the levels of lipid reactive oxygen species(ROS),malondialdehyde(MDA),total iron,and Fe^(2+)in gastric cancer cells,and promoted cell apoptosis(all P<0.05),compared to the control group.The combination of CB and erastin enhanced ferroptosis,while ferrostatin-1 treatment suppressed CB-induced ferroptosis in gastric cancer cells.Mechanistically,CB inhibited the expression of SUMO1,reduced the SUMOylation of HIF1α,and consequently suppressed its expression.The ferroptosis induced by CB in gastric cancer cells could be reversed by overexpression of SUMO1.In vivo experiments confirmed that CB inhibited tumor growth and induced ferroptosis in gastric cancer cells.Conclusion CB induces ferroptosis in gastric cancer cells by in

关 键 词：华蟾素 胃癌 缺氧诱导因子1Α 小泛素样修饰蛋白 铁死亡 细胞凋亡 

分 类 号：R284.1[医药卫生—中药学] R341[医药卫生—中医学] R329.25R591.1R735.2