摘要
目的基于肿瘤蛋白53(p53)/溶质载体家族7成员11(SLC7A11)/谷胱甘肽过氧化物酶4(GPX4)通路探讨去甲斑蝥素诱导卵巢癌细胞铁死亡的作用机制。方法培养人卵巢癌细胞系SKOV3,采用CCK-8法与EdU染色检测细胞增殖能力;划痕实验和Transwell小室实验检测SKOV3细胞的侵袭和迁移能力;试剂盒检测细胞内Fe^(2+)离子、谷胱甘肽(GSH)、丙二醛(MDA)含量,荧光显微镜观察细胞内活性氧(ROS)水平;透射电镜观察细胞线粒体超微形态;Western blot检测细胞p53、SLC7A11、GPX4蛋白表达水平。结果去甲斑蝥素呈浓度依赖性抑制SKOV3细胞活力,IC_(50)值为21.91μmol/L,显著抑制卵巢癌细胞的增殖、迁移及侵袭;显著升高Fe^(2+)水平,降低GSH水平,升高MDA和ROS水平,并使线粒体体积缩小、双层膜密度增加和线粒体嵴减少;显著升高p53蛋白表达水平,降低SLC7A11、GPX4蛋白表达水平。并且Fer-1可部分逆转去甲斑蝥素对SKOV3细胞增殖、迁移及侵袭的调控作用。结论去甲斑蝥素通过调控p53/SLC7A11/GPX4信号通路,促进细胞铁死亡,抑制卵巢癌增殖、迁移及侵袭。
Objective The mechanism of norcantharidin-induced ferroptosis in ovarian cancer cells was investigated based on the tumor protein 53(p53)/solute carrier family 7 member 11(SLC7A11)/glutathione peroxidase 4(GPX4)pathway.Methods Human ovarian cancer cell line SKOv3 was cultured and the proliferation of cells was detected by CCK-8 method and EdU staining.The invasion and migration ability of SKOV3 cells were detected by wound healing and Transwell assay.The contents of Fe^(2+)ion,glutathione(GSH)and malondialdehyde(MDA)were detected by the kit,and the levels of intracellular reactive oxygen species(ROS)were observed by fluorescence microscope.The ultrastructure of mitochondria was observed by transmission electron microscope.The expressions of p53,SLC7A11 and GPX4 were detected by Western blot.Results The norcantharidin inhibited SKOV3 cell viability in a concentrationdependent manner with IC_(50) value of 21.91μmol/L,which significantly inhibited the proliferation,migration and invasion of ovarian cancer cells.The level of Fe2 was significantly increased,the level of GSH level was decreased,the levels of MDA and ROS were increased,mitochondrial volume was reduced,double membrane density was increased and mitochondrial ridge was decreased.The expression of p53 protein was significantly increased,and the expressions of SLC7A11 and GPX4 were decreased.And Fer-1 could partially reverse the regulation of norcantharidin on the proliferation,migration and invasion of SKOV3 cells.Conclusion Norcantharidin may promote ferroptosis by regulating p53/SLC7A11/GPX4 signaling pathway to inhibit the proliferation,migration and invasion of ovarian cancer.
作者
魏晶
李卓
WEI Jing;LI Zhuo(Ward of Gynecological Radiotherapy,Liaoning Cancer Hospital,Shenyang 110044,China)
出处
《解剖科学进展》
2025年第2期241-244,249,共5页
Progress of Anatomical Sciences
基金
新锐肿瘤支持治疗课题研究项目(cphcf-2022-174)。
