下调SCD1表达增敏RSL3诱导铁死亡抑制膀胱癌细胞生长

Down-regulating SCD1 expression sensitizes RSL3 induced ferroptosis and inhibited bladder cancer cell growth

目的:探究SCD1蛋白表达对铁死亡激活剂RSL3诱导膀胱癌细胞铁死亡能力的作用及机制。方法:SCD1小干扰RNA(siRNA)、特异性SCD1抑制剂(A9)处理或联合RSL3处理T24细胞;CCK-8、克隆集落形成实验检测抑制SCD1表达或联合RSL3对细胞增殖及克隆集落形成能力的影响;丙二醛(MDA)及脂质过氧化物(LPO)含量测定分析下调SCD1对RSL3诱导膀胱癌细胞T24铁死亡能力的影响;最后通过蛋白质印迹检测谷胱甘肽过氧化酶4(GPX4)表达。结果:siRNA沉默及A9处理均降低膀胱癌细胞T24增殖及克隆集落形成能力;相对单用组,两种抑制SCD1方法联合RSL3进一步增加膀胱癌细胞MDA及LPO含量,并且增加对增殖及克隆集落形成的抑制程度。此外,抑制SCD1增强RSL3对GPX4蛋白表达的抑制作用。结论:下调SCD1通过增强RSL3对GPX4蛋白表达的抑制,诱发铁死亡发挥抑制膀胱癌细胞生长的作用。

Objective To explore the effect and mechanism of SCD1 protein expression on the ability of ferroptosis activator RSL3 to induce ferroptosis in bladder cancer cells.Method T24 cells were treated with SCD1 small interfering RNA(siRNA),specific SCD1 inhibitor(A9)or combined with RSL3;The effects of inhibiting SCD1 expression or combining with RSL3 on cell proliferation and clonogenesis were detected by CCK-8 and clonogenesis assay;Malondialdehyde(MDA)and lipid peroxides(LPO)were determined to investigate the effect of down-regulated SCD1 on the ability of RSL3 to induce ferroptosis in bladder cancer cells T24;Finally,glutathione peroxidase 4(GPX4)expression was detected by western blot.Results Both siRNA silencing and A9 treatment decreased the proliferation and colony formation ability of bladder cancer cells.Compared with the single treatment group,The two SCD1 inhibition methods combined with RSL3 further increased the MDA and LPO content in the cells,and enhanced the degree of inhibition on the proliferation and clonal colony formation of bladder cancer cells.In addition,inhibition of SCD1 enhanced the inhibitory effect of RSL3 on GPX4 protein expression.Conclusion Down-regulation of SCD1 can inhibit the growth of bladder cancer cells by enhancing the inhibition of GPX4 protein expression by RSL3 and inducing ferroptosis.