蒽醌修饰物4X触发内质网应激诱导卵巢癌细胞死亡模式研究  

Study on the death mode of ovarian cancer cells induced by anthraquinone modifier 4X through triggering endoplasmic reticulum stress

作　　者：许淑妹 杨盈盈 赵英丹 陈强健 张秋萍 侯华新[3] 黎丹戎[1,2] Xu Shumei;Yang Yingying;Zhao Yingdan;Chen Qiangjian;Zhang Qiuping;Hou Huaxin;Li Danrong(College of Oncology,Guangxi Medical University,Nanning 530021,China;Life Sciences Institute,Guangxi Medical University,Nanning 530021,China;Pharmaceutical College,Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学肿瘤医学院,南宁530021 [2]广西医科大学生命科学研究院,南宁530021 [3]广西医科大学药学院,南宁530021

出　　处：《广西医科大学学报》2023年第6期922-929,共8页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.82260714);广西自然科学基金资助项目(No.2020GXNSFDA238016);区域性高发肿瘤早期防治研究教育部重点实验室自主课题项目(No.GKE-ZZ202237)。

摘　　要：目的:探究蒽醌修饰物4X诱导卵巢癌细胞SKOV3和顺铂(DDP)耐药卵巢癌细胞SKOV3/DDP的死亡模式和作用机制。方法:分别将SKOV3细胞和SKOV3/DDP细胞分为对照组和不同浓度(2μmol/L、4μmol/L和8μmol/L)蒽醌修饰物4X组。采用MTT法检测细胞增殖率,苏木精-伊红(HE)染色观察细胞形态,线粒体绿色荧光探针观察线粒体形态,透射电镜观察细胞超微结构,western blotting法检测内质网应激相关蛋白(GRP78、PERK)、凋亡蛋白(CHOP)和铁死亡关键蛋白(GPX4)的表达。加入凋亡抑制剂Z-VAD-FMK,与蒽醌修饰物4X共处理,用流式细胞术检测细胞凋亡率。结果:蒽醌修饰物4X作用48 h后,SKOV3和SKOV3/DDP细胞增殖均明显受到抑制(P<0.05)。经蒽醌修饰物4X处理后SKOV3和SKOV3/DDP细胞质出现明显空泡,且部分空泡累及细胞核,SKOV3和SKOV3/DDP均有明显的线粒体损伤;经蒽醌修饰物4X处理后细胞线粒体固缩,膜密度增高,嵴消失。与对照组相比,蒽醌修饰物4X组SKOV3和SKOV3/DDP细胞凋亡率升高(P<0.05),联用ZVAD-FMK可抑制蒽醌修饰物4X对细胞凋亡的促进作用(P<0.05)。蒽醌修饰物4X可不同程度地下调SKOV3和SKOV3/DDP细胞GRP78、PERK、ATF4和CHOP的表达,上调GPX4表达(均P<0.05)。结论:蒽醌修饰物4X可能在诱导内质网应激的同时诱导SKOV3和SKOV3/DDP细胞发生凋亡和铁死亡,从而发挥抗卵巢癌作用。Objective:To investigate the death mode and mechanism of ovarian cancer cell SKOV3 and cisplatin-resistant ovarian cancer cell SKOV3/DDP induced by anthraquinone modifier 4X.Methods:SKOV3 and SKOV3/DDP cells were divided into control group and different concentrations(2μmol/L,4μmol/L and 8μmol/L)of anthraquinone modifier 4X group,respectively.The cell proliferation rate was detected by MTT assay,the cell morphology was observed by hematoxylin-eosin(HE)staining,the mitochondrial morphology was observed by mito-trayer-green fluorescence probe,and the cell ultrastructure was observed by transmission electron microscopy.The expressions of endoplasmic reticulum stress-related proteins(GRP78,PERK),apoptosis protein(CHOP)and ferroptosis key protein(GPX4)were detected by western blotting.Apoptosis inhibitor Z-VAD-FMK was added and co-treated with anthraquinone modifier 4X,and the apoptosis rate of cells was detected by flow cytometry.Results:After treatment with anthraquinone modifier 4X for 48 hours,the proliferation of SKOV3 and SKOV3/DDP cells was significantly inhibited(P<0.05).After treatment with anthraquinone modifier 4X,there were obvious vacuoles in the cytoplasm of SKOV3 and SKOV3/DDP,and some vacu-oles involved the nucleus.Mitochondrial damage was observed in both SKOV3 and SKOV3/DDP.After treatment with anthraquinone modifier 4X,the mitochondria of the cells were shrunk,the membrane density increased,and the ridge disappeared.Compared with the control group,the apoptosis rate of SKOV3 and SKOV3/DDP cells in the anthraquinone modifier 4X group increased(P<0.05),and the promoting effect of anthraquinone modifier 4X on cell apoptosis was inhibited by adding Z-VAD-FMK(P<0.05).Anthraquinone modifier 4X could upregulate the expressions of GRP78,PERK,ATF4 and CHOP in SKOV3 and SKOV3/DDP cells,and downregulate the expression of GPX4 to varying degrees(all P<0.05).Conclusion:Anthraquinone modifier 4X May induce endoplasmic reticulum stress and the apoptosis and ferroptosis in SKOV3 and SKOV3/DDP cells,thus playing

关 键 词：卵巢癌 蒽醌修饰物 内质网应激 铁死亡 凋亡 

分 类 号：R737.31[医药卫生—肿瘤]