雄激素受体在去势抵抗性前列腺癌细胞铁死亡中的作用研究

The role of androgen receptor in ferroptosis of castration-resistant prostate cancer cells

目的探讨雄激素受体(AR)在去势抵抗性前列腺癌(CRPC)细胞铁死亡中的作用。方法将CRPC细胞系22Rv1和VCaP设置对照组、erastin (10μM)组和erastin+Lip(1μM)组,噻唑蓝(MTT)检测细胞活力,流式细胞仪测定可变铁池(LIP),生化法检测谷胱甘肽(GSH)和丙二醛(MDA)水平,蛋白质印迹法(Western blot)检测AR和谷胱甘肽过氧化酶4(GPX4)蛋白表达,实时定量聚合酶链式反应(qPCR)分析AR的m RNA表达。结果铁死亡诱导剂Erastin以剂量依赖和时间依赖的方式抑制22Rv1和VCaP的细胞活力(P<0.01),特异性铁死亡抑制剂阻止Erastin诱导的细胞生长抑制(P<0.001),与对照组相比,铁死亡诱导剂Erastin促进铁死亡相关性LIP增加,GPX4表达和GSH水平下调,而MDA显著增加,而铁死亡抑制剂Lip使用后逆转(P<0.001)。Erastin降低了AR的表达水平,而Lip则上调了AR的蛋白表达水平,AR的过表达逆转Erastin的细胞生长抑制作用(P<0.01)。结论 AR能逆转铁死亡诱导剂Erastin诱导的细胞生长抑制作用,在CRPC细胞铁死亡中起着重要作用。

Objective To explore the role of androgen receptor(AR) in castration-resistant prostate cancer(CRPC) cell ferroptosis. Methods CRPC cells 22 Rv1 and VCaP were divided into three groups including the control group, group erastin(10 μM) and group erastin + Lip(1 μM). Cell viability was measured by Methylthiazolyldiphenyl-tetrazolium bromide(MTT), labile iron pool(LIP) was detected by flow cytometry, and glutathione(GSH) and Malondialdehyde(MDA) levels were analyzed by biochemical method. The protein expressions of AR and glutathione peroxidase 4(GPX4) were assessed by western blot, and m RNA expression of AR by Quantitative Real-Time Polymerase Chain Reaction(qPCR). Results Ferroptosis inducer Erastin obviously inhibited the cell viability of 22 Rv1 and VCaP cells in a dose-dependent and time-dependent manner(P<0.01). The specific ferroptosis inhibitor prevented Erastin-induced cell growth inhibition(P<0.001).Compared with the control group, ferroptosis inducer Erastin promoted the increase of ferroptosis-related LIP, downregulated GPX4 and GSH expressions, but increased MDA level significantly. Ferroptosis inhibitor Lip reversed these changes described above(P<0.001). Erastin decreased the expression of AR, while Lip increased the expression of AR, and AR overexpression reversed Erastin-induced cell growth inhibitio(P<0.01). Conclusion AR can reverse Erastin-induced cell growth inhibition and play an important role in the iron death of CRPC cells.