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
陈国庆 1董倩男 1杨锐 1高瑛 1刘人嘉 1袁林 2向阳 3吴昊4
作者信息
- 1. 湖北恩施学院医学部,湖北 恩施 445000
- 2. 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
- 3. 湖北恩施学院医学部,湖北 恩施 445000;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;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
- 4. 湖北恩施学院医学部,湖北 恩施 445000;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
- 折叠
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200 μg/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.
Abstract
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 200 μg/mL)PHSTF groups,ferroptosis inhibitor liprox-statin-1(Lip-1)group,and high-dose PHSTF combined with Lip-1 group,each cultured in corresponding media.Cell via-bility 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 em-ployed 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 epichlorohy-drin-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-de-pendent manner(P＜0.01),and the cell migration and invasion were also reduced(P＜0.01),along with a significant de-crease 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Key words
Total flavonoids of Pterocarya hupehensis Skan/non-small-cell lung cancer/ferroptosis/Keap-1/Nrf2/HO-1 pathway引用本文复制引用
基金项目
国家自然科学基金资助项目(81960776)
湖北省自然科学基金一般面上项目(2022CFB515)
湖北省教育厅科学研究指导性项目(B2022495)
恩施州土家族苗族自治州科技局资助项目(JCY20211000040)
出版年
2024
国家科技期刊平台
NSTL
维普
万方数据