百里醌通过诱导铁死亡调控人肺腺癌A549/DDP细胞顺铂的耐药性

Thymoquinone regulates cisplatin resistance of human lung adenocarcinoma A549/DDP cells byinducing ferroptosis

目的:研究百里醌对A549/DDP(人肺腺癌顺铂耐药细胞)细胞顺铂耐药性的影响,并探讨其作用机制。方法:不同浓度百里醌或者顺铂处理A549/DDP细胞,采用CCK8和平板克隆实验检测细胞增殖能力;流式细胞术检测细胞凋亡;流式细胞术检测不同分组中细胞ROS水平;Western blot检测SLC7A11、GPX4、ACSL4等铁死亡相关蛋白的表达。结果:百里醌对A549/DDP细胞的增殖具有抑制作用,并呈剂量和时间依赖性。顺铂联合百里醌与单纯应用顺铂组相比,A549/DDP细胞增殖能力减弱,凋亡率升高。此外,与顺铂单独处理组相比,顺铂联合使用百里醌后,细胞中ROS水平明显升高,ACSL4水平升高,SLC7A11和GPX4蛋白水平明显降低。在加入铁死亡抑制剂Ferrostatin-1后可逆转A549/DDP细胞中ROS水平以及ACSL4、SLC7A11和GPX4等相关蛋白的变化,差异具有统计学意义。结论:百里醌通过诱导铁死亡调控人肺腺癌A549/DDP细胞顺铂的耐药性。

Objective:To investigate the effects of thymoquinone on the cisplatin resistance of human lung adenocarcinoma A549/DDP cells and its mechanism.Methods:A549/DDP cells were treated with different concentrations of TQ or cisplatin,and cell proliferation ability of cells was detected by CCK8 assay and colony forming assay.Cell apoptosis was observed flow cytometry.ROS levels in different group were determined by flow cytometry.Western blot was used to detect the ferroptosis related markers SLC7A11,GPX4 and ACSL4.Results:Thymoquinone inhibited the proliferation of A549/DDP cells in a dose-and time-dependent manner.Thymoquinone combined with cisplatin decreased the proliferation and increased the apoptosis significantly in A549/DDP cells,compared with cisplatin alone group.Thymoquinone combined with cisplatin increased ROS level,ACSL4 level,and decreased SLC7A11 as well as GPX4,which could be reversed by ferroptosis inhibitor Ferrostatin-1.Conclusion:Thymoquinone regulates cisplatin resistance of human lung adenocarcinoma A549/DDP cells through inducing ferroptosis.