摘要
目的 探讨中药靛玉红衍生物E804对非小细胞肺癌(NSCLC)系A549细胞增殖和迁移的影响,并阐明Nrf2-HO-1/GPX4信号轴可能的作用机制。方法 以肺癌A549细胞为细胞模型。采用MTT和细胞划痕实验观察0、10μmol/L E804和10μmol/L E804+不同特异性抑制剂(Nec-1、CQ、Z-VAD、DFO、Fer-1和Lip-1)组细胞的增殖和迁移能力;采用DCFH-DA荧光探针法检测0、2.5、5和10μmol/LE804组细胞内活性氧(ROS)含量,比色法检测二价铁离子(Fe^(2+))含量,分光光度法检测还原型谷胱甘肽(GSH)含量,微量法检测丙二醛(MDA)含量;Western blot法检测0、2.5、5和10μmol/L E804组细胞SLC7A11、Transferrin、GPX4、SLC40A1、Nrf2和HO-1蛋白表达水平。结果 与对照组(0μmol/LE804)比较,2.5、5和10μmol/L E804升高细胞内ROS、Fe^(2+)和MDA水平,降低细胞内GSH含量(P<0.01),同时降低SLC7A11、GPX4、SLC40A1、Nrf2和HO-1蛋白表达水平(P<0.01),升高Transferrin表达水平(P<0.05)。与单独使用10μmol/LE804组比较,细胞凋亡抑制剂(Z-VAD)组和细胞铁死亡抑制剂(DFO、Fer-1和Lip-1)组能够部分逆转E804对A549细胞的增殖和迁移抑制(P<0.01)。结论 E804可抑制A549细胞增殖和迁移,并诱发铁死亡,其机制可能与抑制Nrf2-HO-1/GPX4信号轴有关。
Objective To investigate the effects of indirubatin derivative E804 on proliferation and migration of non-small cell lung cancer(NSCLC) A549 cells,and to elucidate the possible mechanism of Nrf2-HO-1/GPX4 pathway.Methods Lung cancer A549 cells were used as the cell model.The proliferation and migration of different specific inhibitors(Nec-1,CQ,Z-VAD,DFO,Fer-1 and Lip-1) in 0,10 μmol/L E804 and 10 μmol/L E804+ groups were observed by MTT and cell scratch assay.The contents of reactive oxygen species(ROS) were detected by DCFH-DA fluorescence probe method,the contents of Fe^(2+) were detected by colorimetric method,the contents of reduced glutathione(GSH) were detected by spectrophotometry,and the contents of malondialdehyde(MDA) were detected by micromethod.The expression levels of SLC7A11,Transferrin,GPX4,SLC40A1,Nrf2 and HO-1 were detected by Western blot in cells of 0,2.5,5 and 10 μmol/L E804 groups.Results Compared with the control group(0 μmol/L E804),2.5,5 and 10 μmol/L E804 significantly increased intracellular ROS,Fe^(2+) and MDA levels,and decreased intracellular GSH content(P<0.01).Meanwhile,the expression levels of SLC7A11,GPX4,SLC40A1,Nrf2 and HO-1 significantly decreased(P<0.01),and the expression level of Transferrin increased(P<0.05).Compared with the 10 μmol/L E804 group alone,the apoptosis inhibitor(Z-VAD) group and the ferroptosis inhibitor(DFO,Fer-1 and Lip-1) group could significantly reverse the inhibition of proliferation and migration of A549 cells by 10 μmol/L E804(P<0.01).Conclution E804 can induce ferroptosis and inhibit the proliferation and migration of A549 cells,which may be related to the inhibition of Nrf2-HO-1/GPX4 pathway.
作者
袁育珺
曹华华
赵敏
罗宇慧
张素梅
Yuan Yujun;Cao Huahua;Zhao Min;Luo Yuhui;Zhang Sumei(Clinical Laboratory,The Affiliated Hospital of Jiujiang University,Jiujiang 332000;Clinical Laboratory,The Traditional Chinese Medicine of Jiujiang,Jiujiang 332000;Pathology,The People's Hospital of Nanchang,Nanchang 330000;Dept of Molecular Biology and Biochemistry,Anhui Medical University,Hefei 230032)
出处
《安徽医科大学学报》
CAS
北大核心
2024年第2期331-335,343,共6页
Acta Universitatis Medicinalis Anhui
基金
江西省中医药管理局科技计划项目(编号:2023B1322)
江西省卫生健康委科技计划项目(编号:202410703)。
