青蒿琥酯通过p53/SLC7A11/GPX4轴诱导人骨肉瘤细胞铁死亡  

Artesunate induces ferroptosis of human osteosarcoma cells through p53/SLC7A11/GPX4 axis

作　　者：李明 蒋江梅 颜丽君 朱青 吴太鼎 陈龙菊 LI Ming;JIANG Jiangmei;YAN Lijun;ZHU Qing;WU Taiding;CHEN Longju(Hubei Provincial Key Laboratory of Occurrence and Intervention of Rheumatic Diseases,Health Scinence Center of Hubei Minzu University,Enshi 445000,China)

机构地区：[1]风湿性疾病发生与干预湖北省重点实验室,湖北民族大学医学部,湖北恩施445000

出　　处：《中国病理生理杂志》2024年第9期1606-1611,共6页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81260192);风湿性疾病发生与干预湖北省重点实验室2024年度开放基金项目(No.PT022405)。

摘　　要：目的:探讨青蒿琥酯(Art)对骨肉瘤的促铁死亡作用及其机制。方法:将人骨肉瘤U-2 OS细胞分为4组:正常对照组、ferrostatin-1(Fer-1)组、Art组和Art+Fer-1组。CCK-8法检测细胞活力;生化方法检测细胞ROS、Fe^(2+)和GSH水平;透射电镜观察细胞线粒体形态;RT-qPCR检测p53、溶质载体家族7成员11(SLC7A11)和谷胱甘肽过氧化生物酶4(GPX4)的mRNA水平;Western blot检测p53、SLC7A11和GPX4蛋白水平。结果:Art显著抑制了U-2 OS细胞的生长。Art通过促进Fe^(2+)的积累和ROS的形成、抑制GSH的产生,诱发了OS细胞铁死亡,而铁死亡抑制剂Fer-1则抑制了U-2 OS细胞铁死亡。此外,Art还改变了U-2 OS细胞的线粒体形态,表现为线粒体缩小,线粒体膜密度增高,线粒体嵴减少。Art抑制了铁死亡通路上SLC7A11和GPX4 mRNA及蛋白的表达、上调了铁死亡上游调控因子p53的表达,从而诱导铁死亡。结论:Art能通过p53/SLC7A11/GPX4通路诱导骨肉瘤细胞铁死亡。AIM:To explore the promotion of ferroptosis by artesunate(Art)and its underlying mechanism in osteosarcoma.METHODS:Human osteosarcoma U-2 OS cells were divided into 4 groups:control,ferrostatin-1(Fer-1),Art,and Art+Fer-1 groups.Cell viability was measured by CCK-8 assay,while reactive oxygen species(ROS),Fe^(2+),and glutathione(GSH)levels were measured using biochemical assays.Mitochondrial morphology was evaluated by transmission electron microscopy.The mRNA and protein levels of p53,solute carrier family 7 member 11(SLC7A11)and glutathione peroxidase 4(GPX4)were determined by RT-qPCR and Western blot,respectively.RESULTS:It was found that Art significantly reduced the growth of U-2 osteosarcoma cells,as well as inducing ferroptosis by promoting the accumulation of Fe^(2+)and ROS and reducing GSH levels,while the ferroptosis inhibitor ferrostatin-1 inhibited ferroptosis.It was also observed that Art affected mitochondrial morphology,resulting in smaller mitochondria,higher mitochondrial membrane density,and reduced numbers of cristae.Treatment with Art also downregulated both the mRNA and protein expression of the ferroptosis-associated genes SLC7A11 and GPX4,while upregulating expression of p53,an upstream regulator of ferroptosis,thus inducing ferroptosis.CONCLUSION:Artesunate induces ferroptosis in osteosarcoma cells through the p53/SLC7A11/GPX4 signaling pathway.

关 键 词：骨肉瘤 铁死亡 p53/SLC7A11/GPX4信号通路 青蒿琥酯 

分 类 号：R738[医药卫生—肿瘤] R363.2[医药卫生—临床医学] Q255[生物学—细胞生物学]