蒽醌修饰物KA-4s抑制SKOV3/DDP细胞增殖并诱导铁死亡  

Anthraquinone derivative KA-4s inhibiting the proliferation and inducing ferroptosis of SKOV3/DDP cells

作　　者：赵英丹 李欣晓 许淑妹 陈强健 杨盈盈 侯华新[3] 黎丹戎[1,2] ZHAO Yingdan;LI Xinxiao;XU Shumei;CHEN Qiangjian;YANG Yingying;HOU Huaxin;LI Danrong(College of Oncology,Guangxi Medical University,Nanning 530021,China;Life Sciences Institute,Guangxi Medical University,Nanning 530021,China;College of Pharmacy,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》2024年第3期356-363,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:探讨蒽醌修饰物KA-4s在体外对人顺铂耐药卵巢癌SKOV3/DDP细胞增殖的影响和诱导细胞铁死亡的机制。方法:将SKOV3/DDP细胞分为对照组和不同浓度(2μmol/L、5μmol/L和7μmol/L)的蒽醌修饰物KA-4s组。采用MTT法检测单药KA-4s和顺铂(DDP)对SKOV3/DDP细胞活力的影响,划痕实验检测细胞的迁移能力,线粒体绿色荧光探针(Mito-Tracker Green)检测线粒体形态改变,透射电镜观察线粒体超微结构。以铁死亡诱导剂RSL3为阳性对照组,用DCFA-DA荧光探针检测细胞内活性氧(ROS)水平,比色法检测细胞内总铁蛋白含量,western blotting检测铁死亡相关蛋白GPX4、FSP1的表达情况。结果:KA-4s和顺铂作用48 h后,卵巢癌SKOV3/DDP细胞增殖均明显受到抑制,相比顺铂,KA-4s的抑制作用更强(P<0.001),并能抑制细胞迁移。经KA-4s处理后线粒体受损,线粒体结构改变,膜密度增大,嵴减少甚至消失。与空白对照组相比,阳性RSL3组和KA-4s组SKOV3/DDP细胞内ROS水平升高(均P<0.001),5μmol/L KA-4s组总铁离子含量显著升高(P<0.001),卵巢癌SKOV3/DDP细胞中GPX4、FSP1蛋白的表达均降低(P<0.01),但正常卵巢IOSE80细胞中GPX4表达均无改变。结论:KA-4s能抑制SKOV3/DDP细胞增殖、迁移并诱导细胞铁死亡。Objective:To investigate the effect of anthraquinone derivative KA-4s on the proliferation of human cisplatin-resistant ovarian cancer SKOV3/DDP cells in vitro and the mechanism of inducing ferroptosis.Methods:SKOV3/DDP cells were divided into control group and different concentrations(2μmol/L,5μmol/L and 7μmol/L)of anthraquinone derivative KA-4s groups.The effects of KA-4s and cisplatin(DDP)alone on the viability of SKOV3/DDP cells were determined by MTT assay,the migration ability of SKOV3/DDP cells was detected by scratch test,the mitochondrial morphological changes were detected by mitochondrial green fluorescent probe(Mito-Tracker Green),and the mitochondrial ultrastructure was observed by transmission electron microscopy.Ferroptosis inducer RSL3 was used as a positive control group,intracellular reactive oxygen species(ROS)levels were detected by DCFA-DA fluorescent probe,the total intracellular ferritin content was detected by colorimetry,and the expression of ferroptosis-related proteins GPX4 and FSP1 was detected by western blotting.Results:After treatment with KA-4s and DDP for 48 h,the proliferation of ovarian cancer SKOV3/DDP cells was significantly inhibited,and KA-4s had a stronger inhibitory effect than DDP(P<0.001),and could inhibit cell migration.After KA-4s treatment,mitochondria were damaged,mitochondrial structure changed,membrane density increased,and ridge decreased or even disappeared.Compared with the blank control group,the intracellular ROS levels of the positive RSL3 group and KA-4s group were increased(all P<0.001),the total ironcontent in 5μmol/L KA-4s group was significantly increased(P<0.001),and the expression of GPX4 and FSP1 proteins in ovarian cancer SKOV3/DDP cells was decreased(P<0.01),but GPX4 expression in the normal ovarian IOSE80 cells was unchanged.Conclusion:KA-4s can inhibit the proliferation,migration and induce ferroptosis of SKOV3/DDP cells.

关 键 词：蒽醌修饰物 SKOV3/DDP细胞 铁死亡 谷胱甘肽过氧化物酶4 活性氧 

分 类 号：R963[医药卫生—微生物与生化药学]