湖北枫杨总黄酮对非小细胞肺癌A549细胞迁移、侵袭和铁死亡的影响  

Effects of total flavonoids of Pterocarya hupehensis Skan on migration,invasion and ferroptosis of non-small-cell lung cancer A549 cells

作　　者：陈国庆 董倩男 杨锐 高瑛 刘人嘉 袁林 向阳 吴昊 CHEN Guoqing;DONG Qiannan;YANG Rui;GAO Ying;LIU Renjia;YUAN Lin;XIANG Yang;WU Hao(Medical School of Hubei Enshi University,Enshi 445000,China;Enshi Huiyi Hospital for Rheumatic Diseases,Enshi 445000,China;Hubei Provincial Key Laboratory of Occurrence and Intervention of Rheumatic Diseases,Hubei Minzu Uni-versity,Enshi 445000,China)

机构地区：[1]湖北恩施学院医学部,湖北恩施445000 [2]湖北恩施学院附属恩施慧宜中西医结合风湿医院,湖北恩施445000 [3]湖北民族大学风湿性疾病发生与干预湖北省重点实验室,湖北恩施445000

出　　处：《中国病理生理杂志》2024年第2期274-281,共8页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81960776);湖北省自然科学基金一般面上项目(No.2022CFB515);湖北省教育厅科学研究指导性项目(No.B2022495);恩施州土家族苗族自治州科技局资助项目(No.JCY20211000040)。

摘　　要：目的:探究湖北枫杨总黄酮对非小细胞肺癌A549细胞迁移与侵袭及铁死亡的影响及机制。方法:将A549细胞分为:对照组,低、中、高剂量(100、150和200μg/mL)枫杨总黄酮组,铁死亡抑制剂liproxstatin-1(Lip-1)组,Lip-1+高剂量枫杨总黄酮组。采用CCK-8法检测细胞活力;划痕实验和Transwell实验检测细胞迁移及侵袭能力;谷胱甘肽(GSH)试剂盒检测细胞脂质过氧化水平;RT-qPCR检测细胞溶质载体家族7成员11(SLC7A11)和谷胱甘肽过氧化物酶4(GPX4)的mRNA水平;Western blot检测SLC7A11、GPX4、Kelch样环氧氯丙烷相关蛋白1(Keap1)、核因子E2相关因子2(Nrf2)和血红素加氧酶1(HO-1)的蛋白表达水平。结果:与对照组相比,各剂量枫杨总黄酮组A549细胞活力降低(P<0.01),迁移及侵袭能力降低(P<0.01),GSH含量显著降低(P<0.01),SLC7A11和GPX4 mRNA及蛋白表达水平显著降低(P<0.01),Nrf2和HO-1蛋白表达水平显著降低(P<0.01),Keap-1蛋白表达水平升高(P<0.01);加入铁死亡抑制剂后,与高剂量枫杨总黄酮组相比,A549细胞活力显著升高(P<0.01),SLC7A11、GPX4、Nrf2、HO-1蛋白表达水平升高,Keap-1蛋白表达水平下降(P<0.01)。结论:湖北枫杨总黄酮能抑制A549细胞迁移与侵袭并且通过调控Keap-1/Nrf2/HO-1通路诱导肺癌A549细胞铁死亡。AIM:To investigate the impact of total flavonoids of Pterocarya hupehensis Skan(PHSTF)on the migration,invasion,and ferroptosis of non-small-cell lung cancer A549 cells.METHODS:The A549 cells were divided into control group,low-,medium-and high-dose(100,150 and 200μg/mL)PHSTF groups,ferroptosis inhibitor liproxstatin-1(Lip-1)group,and high-dose PHSTF combined with Lip-1 group,each cultured in corresponding media.Cell viability was assessed using the CCK-8 assay,while cell migration and invasion abilities were determined through scratch and Transwell assays.Cell lipid peroxidation levels were measured using the glutathione(GSH)assay kit.RT-qPCR was employed to assess the mRNA expression of solute carrier family 7 member 11(SLC7A11)and glutathione peroxidase 4(GPX4),while Western blot was utilized to examine the protein expression of SLC7A11,GPX4,Kelch-like epichlorohydrin-associated protein-1(Keap-1),nuclear factor E2-related factor 2(Nrf2)and heme oxygenase-1(HO-1).RE⁃SULTS:Compared with control group,PHSTF significantly diminished the viability of A549 cells in a time-and dose-dependent manner(P<0.01),and the cell migration and invasion were also reduced(P<0.01),along with a significant decrease in GSH level(P<0.01).Treatment with PHSTF inhibited the mRNA and protein expression levels of ferroptosis-re-lated proteins,including SLC7A11 and GPX4(P<0.01),suppressed the protein expression of Nrf2 and HO-1(P<0.01),and enhanced the expression of Keap-1(P<0.01).The Lip-1 partially restored the decrease in cell viability in-duced by PHSTF(P<0.01),significantly up-regulated the protein expression levels of SLC7A11,GPX4,Nrf2 and HO-1,and suppressed the protein expression of Keap-1(P<0.01).CONCLUSION:Total flavonoids of Pterocarya hupehen⁃sis Skan can inhibit the migration and invasion of non-small-cell lung cancer A549 cells,and induce the cell ferroptosis by regulating the Keap-1/Nrf2/HO-1 pathway.

关 键 词：湖北枫杨总黄酮 非小细胞肺癌 铁死亡 Keap-1/Nrf2/HO-1通路 

分 类 号：R734.2[医药卫生—肿瘤] R363.2[医药卫生—临床医学]