伊维菌素对去势抵抗性前列腺癌铁死亡的影响  

Effect of ivermectin on ferroptosis of castration-resistant prostate cancer

作　　者：杜永峰 房晓雪 司明贵 马金鹏 苏信旺 李东龙 王科亮[1] DU Yongfeng;FANG Xiaoxue;SI Minggui;MA Jinpeng;SU Xinwang;LI Donglong;WANG Keliang(Fourth Hospital of Harbin Medical University,Harbin 150001,China)

机构地区：[1]哈尔滨医科大学附属第四医院泌尿外科,哈尔滨150001 [2]哈尔滨医科大学附属第四医院中心手术室,哈尔滨150001 [3]甘肃省中心医院泌尿外科

出　　处：《现代泌尿生殖肿瘤杂志》2025年第1期32-40,共9页Journal of Contemporary Urologic and Reproductive Oncology

基　　金：黑龙江省卫生健康委科研课题(2020-082);国家自然科学基金面上项目(82172782);黑龙江省自然科学基金项目(LH2021H040);哈尔滨医科大学“少帅揭榜”项目(HMUMIF-22007)。

摘　　要：目的探讨伊维菌素(IVM)对去势抵抗性前列腺癌(CRPC)增殖的影响及其诱导铁死亡的分子机制。方法体外培养人CRPC细胞系DU145,分别设置对照组、IVM处理组、IVM和铁死亡抑制剂(Fer-1)联合处理组,利用CCK8、丙二醛(MDA)定量分析、细胞内活性氧(ROS)荧光染色及细胞内Fe^(2+)含量测定实验来验证IVM对CRPC增殖和铁死亡的影响。利用Western Blot检测IVM对铁死亡相关蛋白谷胱甘肽过氧化物酶4(GPX4)的影响。利用生物信息学技术筛选GPX4上游转录因子并验证其对CRPC生存期的影响,敲低E2F1检测E2F1对GPX4表达的影响。利用Western Blot检测IVM对E2F1蛋白表达的影响,并通过细胞划痕、CCK8、集落形成、MDA定量分析、ROS荧光染色及细胞内Fe^(2+)含量测定实验来验证E2F1对CRPC迁移、增殖的影响及其与铁死亡之间的关系。结果CCK8、MDA定量分析、ROS荧光染色及细胞内Fe^(2+)含量测定结果显示IVM通过促进铁死亡抑制CRPC增殖(均P<0.05)。IVM显著抑制GPX4蛋白的表达(P<0.01)。成功筛选出GPX4的转录因子E2F1,生存分析显示低表达E2F1可显著延长CRPC患者的生存期(P<0.05),敲低E2F1显著抑制GPX4表达(P<0.05)。IVM抑制E2F1蛋白表达(P<0.05),敲低E2F1显著促进铁死亡抑制CRPC迁移和增殖(均P<0.05)。结论IVM通过下调E2F1抑制GPX4蛋白表达促进CRPC铁死亡。Objective To investigate the effect of ivermectin(IVM)on the proliferation of castration-resistant prostate cancer(CRPC)and the molecular mechanism of ferroptosis.Methods Human CRPC cell line DU145 was cultured in vitro.The control group,IVM group,IVMand Ferrostatin-1(Fer-1)group were set up respectively.CCK8,Malondialdehyde IVM(MDA)quantitative analysis,Reactive Oxygen Species(ROS)fluorescence staining and determination of intracellular Fe^(2+) content were used to verify the effect of IVM on the proliferation and ferroptosis of CRPC.The effect of IVM on ferroptosis related protein glutathione peroxidase 4(GPX4)was detected by Western Blot.Bioinformatics techniques were used to screen the upstream transcription factors of GPX4 and to verify their effects on the survival of CRPC.The effect of E2F1 on GPX4 expression was detected by knocking down E2F1.The effect of IVM on E2F1 protein expression was detected by Western Blot,and the effect of E2F1 on migration,proliferation and ferroptosis of CRPC was verified by cell scratch test,CCK8,colony formation,MDA quantitative analysis,ROS fluorescence staining and determination of intracellular Fe^(2+) content.Results CCK8,MDA quantitative analysis,ROS fluorescence staining and determination of intracellular Fe^(2+) content showed that IVM inhibited the proliferation of CRPC by promoting ferroptosis(all P<0.05).IVM significantly inhibited the expression of GPX4 protein(P<0.05).The transcription factor E2F1 of GPX4 was successfully screened.Survival analysis showed that low expression of E2F1 could significantly prolong the survival time of patients with CRPC(P<0.05).Knocking down E2F1 significantly inhibited the expression of GPX4(P<0.05).IVM inhibited the expression of E2F1 protein(P<0.05).Knocking down E2F1 significantly promotes ferroptosis and inhibits migration and proliferation of CRPC allP<0.05).Conclusions IVM promotes ferroptosis of CRPC by down-regulating E2F1 and inhibiting GPX4 protein expression.

关 键 词：伊维菌素 去势抵抗性前列腺癌 铁死亡 E2F1因子 谷胱甘肽过氧化物酶4 

分 类 号：R73[医药卫生—肿瘤]