去甲斑蝥素通过Nrf2/HO-1通路介导铁死亡抑制小细胞肺癌H446细胞增殖  

Norcantharidin inhibits the proliferation of small cell lung cancer H446 cells via Nrf2/HO-1 pathway mediated ferroptosis

作　　者：苏帆 刘单 邓述恺 SU Fan;LIU Dan;DENG Shukai(Department of Respiratory and Critical Care Medicine,Affiliated Hospital of Southwest Medical University,Sichuan Luzhou 646000,China)

机构地区：[1]西南医科大学附属医院呼吸与危重症医学科,四川泸州646000

出　　处：《现代肿瘤医学》2025年第4期583-589,共7页Journal of Modern Oncology

摘　　要：目的:探讨去甲斑蝥素(norcantharidin,NCTD)对小细胞肺癌H446细胞增殖的影响,研究NCTD影响H446细胞增殖与铁死亡的相关性。方法:采用梯度浓度(0、2、4、8、16、32、64μg/mL)NCTD处理小细胞肺癌H446细胞24、48、72 h后,CCK-8法检测去甲斑蝥素对细胞增殖的影响,计算半数抑制浓度(IC50)用于后续试验。设置对照组、NCTD组、NCTD+Fer-1组,透射电子显微镜观察给药后细胞线粒体超微结构变化,流式细胞术检测细胞内活性氧(reactive oxygen species,ROS)水平,FerroOrange亚铁离子荧光探针试剂盒检测Fe^(2+)水平,Western blot实验检测各组细胞中铁死亡相关蛋白溶质载体家族7成员11(solute carrier family 7 member 11,SLC7A11),谷胱甘肽过氧化物酶4(glutathione peroxidase 4,GPX4),核因子红细胞系2相关因子2(nuclear factor erythroid 2-related factor 2,Nrf2),血红素加氧酶-1(heme oxygenase-1,HO-1)的表达水平。结果:去甲斑蝥素可抑制小细胞肺癌H446细胞增殖,且呈一定的时间和浓度依赖趋势(P<0.05)。透射电镜下可见去甲斑蝥素引起细胞铁死亡典型形态学改变。去甲斑蝥素可上调细胞内Fe^(2+)、ROS水平,而铁死亡抑制性蛋白SLC7A11、GPX4、Nrf2、HO-1的表达均下调(P<0.05);这些指标均可被铁死亡抑制剂Ferrostatin-1(Fer-1)逆转。结论:去甲斑蝥素能够抑制小细胞肺癌H446细胞增殖能力,这一过程可能是通过下调Nrf2/HO-1通路的表达,升高细胞内Fe^(2+)、ROS水平,从而诱导铁死亡实现的。Objective:To investigate the effect of norcantharidin(NCTD)on the proliferation of small cell lung cancer H446 cells,and to study the correlation between the proliferation of H446 cells and ferroptosis.Methods:H446 cells were treated with gradient concentrations of NCTD(0,2,4,8,16,32,64μg/mL)for 24 h,48 h,and 72 h.The effect of NCTD on cell proliferation was detected by CCK-8 assay,and the median inhibitory concentration(IC 50)was calculated for subsequent experiments.The experiment was divided into control group,NCTD group and NCTD+Fer-1 group.The ultrastructural morphological changes of mitochondria were observed by transmission electron microscopy.The level of reactive oxygen species(ROS)in the cells was detected by flow cytometry,and the expression of Fe^(2+)was detected by FerroOrange fluorescent probe kit.Protein expression level of ferroptosis-related recombinant solute carrier family 7 member 11(SLC7A11),glutathione peroxidase 4(GPX4),nuclear factor erythroid 2-related factor 2(Nrf2),and heme oxygenase-1(HO-1)in H446 cells was detected by Western blot.Results:The proliferation of H446 cells was inhibited by NCTD in a time and concentration dependent manner(P<0.05).Typical morphological changes of ferroptosis induced by NCTD were observed under transmission electron microscopy.NCTD up-regulated the intracellular Fe^(2+)and ROS levels,while down-regulated the expression of ferroptosis inhibitory proteins SLC7A11,GPX4,Nrf2 and HO-1(P<0.05).These changes could be reversed by ferroptosis inhibitor Ferrostatin-1(Fer-1).Conclusion:NCTD could inhibit the proliferation of H446 cells,which may induce ferroptosis by down-regulating the expression of Nrf2/HO-1 pathway,increasing intracellular Fe^(2+)and ROS levels.

关 键 词：小细胞肺癌 去甲斑蝥素 铁死亡 Nrf2/HO-1通路 

分 类 号：R734.2[医药卫生—肿瘤]