基于ATF2-Nrf2通路探讨肺岩宁调控非小细胞肺癌铁死亡敏感性的作用机制  

Regulation of Feiyanning on sensitivity of non-small cell lung cancer to ferroptosis based on ATF2-Nrf2 pathway

作　　者：高红芳 周兰 周己扬 沈彬雯 徐振晔[2] GAO Hongfang;ZHOU Lan;ZHOU Jiyang;SHEN Binwen;XU Zhenye(Department of Oncology,Shanghai Yangpu Hospital of Traditional Chinese Medicine,Shanghai 200090,China;DepartmentⅡof Oncology,Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China)

机构地区：[1]杨浦区中医医院肿瘤科,上海200090 [2]上海中医药大学附属龙华医院肿瘤二科,上海200032

出　　处：《上海中医药杂志》2025年第4期64-71,共8页Shanghai Journal of Traditional Chinese Medicine

基　　金：国家中医药管理局全国名老中医药专家传承工作室建设项目(国中医药人教函[2022]75号);上海市杨浦区科委、卫健委中医专项课题(YPZM202107)。

摘　　要：目的探讨肺岩宁对非小细胞肺癌铁死亡敏感性的影响及其机制。方法利用肺岩宁干预铁死亡诱导剂埃拉斯汀(Erastin)诱导的非小细胞肺癌A549细胞铁死亡,通过细胞计数试剂盒(CCK-8)检测细胞活性改变,通过谷胱甘肽(GSH)检测试剂盒、亚铁离子(Fe2+)检测试剂盒、丙二醛(MDA)检测试剂盒分别检测细胞内GSH水平、Fe2+水平及MDA水平改变。通过高内涵成像检测活性氧(ROS)敏感荧光探针标记的细胞内氧化应激改变。通过Western blot法检测激活转录因子2(ATF2)、核因子2相关因子2(Nrf2)、溶质载体家族7成员11(SLC7A11)及谷胱甘肽过氧化物酶4(GPX4)表达情况。结果与对照组比较,低浓度肺岩宁对A549细胞存活率影响差异无统计学意义(P>0.05),而中浓度肺岩宁(P<0.01)及高浓度肺岩宁(P<0.001)可明显降低A549细胞存活率。与各浓度Erastin组比较,肺岩宁联合各浓度Erastin组均可明显降低A549细胞存活率,差异有统计学意义(P<0.05,P<0.01)。与肺岩宁组、Erastin组、Erastin联合肺岩宁组比较,铁死亡抑制剂铁抑素-1(Ferrostatin-1)干预后各组细胞存活率均明显提高,差异有统计学意义(P<0.05,P<0.01,P<0.001)。Western blot检测显示,与对照组比较,Erastin能明显上调A549细胞内ATF2表达(P<0.05),下调Nrf2(P<0.001)、SLC7A11(P<0.05)及GPX4表达(P<0.01)。与对照组比较,肺岩宁可显著下调ATF2(P<0.05)及Nrf2表达(P<0.01),而对SLC7A11及GPX4表达无显著影响(P>0.05)。此外,与对照组比较,Erastin联合肺岩宁可显著下调ATF2(P<0.05)、Nrf2(P<0.001)、SLC7A11(P<0.001)及GPX4表达(P<0.001)。而与Erastin组比较,Erastin联合肺岩宁可显著下调ATF2(P<0.001)、Nrf2(P<0.05)、SLC7A11(P<0.05)及GPX4表达(P<0.05)。结论肺岩宁可以增强非小细胞肺癌细胞对铁死亡诱导剂的敏感性,其机制可能与其调控的ATF2-Nrf2通路有关。Objective To explore the effect of Feiyanning(FYN)on sensitivity of non-small cell lung cancer(NSCLC)to ferroptosis and its mechanism.Methods Erastin-induced NSCLC A549 cells were treated with FYN.Cell viability was examined using cell counting kit-8(CCK-8)assay.Intracellular level of reduced glutathione(GSH),Fe2+and malondialdehyde(MDA)were examined using GSH assay kit,ferrous ion content assay kit and MDA assay kit,respectively.Intracellular reactive oxygen species(ROS)was labeled using ROS-sensitive probe and determined through high-content imaging system.The expressions of activating transcription factor 2(ATF2),nuclear factor E2-related factor 2(Nrf2),solute carrier family 7 member 11(SLC7A11)and glutathione peroxidase 4(GPX4)were examined using Western blot.Results Although low concentration of FYN showed no significant effect on the cell viability of A549 cells,medium and high concentration of FYN significantly decreased the cell viability of A549 cells(P<0.01,P<0.001)compared with that of the control group.Compared with each concentration group of Erastin,FYN combined with each concentration of Erastin significantly reduced the cell viability of A549 cells,and the difference was statistically significant(P<0.05,P<0.01).Compared with FYN group,Erastin group,and Erastin combined with FYN group,the cell viability in each group was significantly improved after the intervention of Ferrostatin-1(P<0.05,P<0.01,P<0.001).Western blot analysis showed that compared with the control group,Erastin could significantly up-regulate ATF2 expression(P<0.05),down-regulate the expressions of Nrf2(P<0.001),SLC7A11(P<0.05)and GPX4(P<0.01)in A549 cells.Compared with the control group,FYN significantly down-regulated the expressions of ATF2(P<0.05)and Nrf2(P<0.01),but had no significant effect on SLC7A11 and GPX4 expressions(P>0.05).In addition,compared with the control group,Erastin combined with FYN significantly down-regulated the expressions of ATF2(P<0.05),Nrf2(P<0.001),SLC7A11(P<0.001)and GPX4(P<0.001).Compared with Erasti

关 键 词：非小细胞肺癌 铁死亡 肺岩宁 作用机制 中药研究 

分 类 号：R73[医药卫生—肿瘤]