皮质酮对Erastin诱导黑色素瘤细胞铁死亡的促进作用

Corticosterone accelerated Erastin-induced ferroptosis in melanoma

目的:探讨皮质酮(corticosterone)对铁死亡激活剂Erastin诱导黑色素瘤细胞铁死亡的影响。方法:单独或联合使用10μmol/L的二甲基亚砜(dimethyl sulfoxide, DMSO)、Erastin、皮质酮处理B16-F10细胞。采用MTT法检测细胞活性,透射电子显微镜观察细胞形态的改变,分别用铁离子比色法、谷胱甘肽(glutathione reduced, GSH)、丙二醛(malondialdehyde, MDA)试剂盒检测细胞内铁离子、GSH和MDA的含量,通过荧光探针DCFH-DA检测细胞内活性氧(reactive oxygen species, ROS)水平。结果:与对照组比较,Erastin组、皮质酮组和Erastin+皮质酮组的细胞活性在处理48、72 h后显著下降(P<0.05)。电镜结果发现Erastin组、皮质酮组、Erastin+皮质酮组发生线粒体缩小,结构紧密,嵴减少或消失,Erastin+皮质酮组甚至出现线粒体肿胀、空泡变。Erastin组、皮质酮组和Erastin+皮质酮组细胞内铁、MDA、ROS含量较对照组显著升高(P<0.05),而Erastin组和Erastin+皮质酮组细胞中GSH水平明显低于对照组(P<0.05),上述变化Erastin+皮质酮组比Erastin组更为明显。结论:皮质酮可促进Erastin诱导黑色素瘤细胞铁死亡。

Objective:To investigate the effect of corticosterone on Erastin-induced ferroptosis in melanoma.Methods:The B16-F10 cells were treated with 10 μmol/L dimethyl sulfoxide(DMSO),Erastin and corticosterone alone or in combination.MTT assay was used to detect the cell activity.Transmission electron microscope was used to observe the cell mitochondrial morphology.The iron, glutathione(GSH) and malondialdehyde(MDA) kit were used to detect the contents of iron, GSH and MDA.And the level of ROS was detected by fluorescent probe DCFH-DA.Results:Compared with the control group, the cell activity of Erastin group, corticosterone group and Erastin+corticosterone group decreased significantly after 48 and 72 hours treatment(P<0.05).We observed that mitochondria were shrunken and compacted with reduction or disappearance of cristae in the Erastin, corticosterone, and Erastin+corticosterone treatment group.Mitochondria in Erastin+corticosterone group even swelled and vacuolated.The contents of iron, MDA and ROS in Erastin group, corticosterone group and Erastin+corticosterone group were significantly higher than those in the control group(P<0.05),while the level of GSH in Erastin group and Erastin+corticosterone group was significantly lower than that in the control group(P<0.05).The changes of above indexes were more obvious in the Erastin+corticosterone group than in the Erastin group.Conclusion:Corticosterone accelerated Erastin-induced ferroptosis in melanoma.