摘要
目的探讨雄黄对食管癌细胞增殖、侵袭及铁死亡的影响。方法不同浓度雄黄(0、10、20、40、60、80、100μmol·L^(-1))或雄黄1/2IC_(50)、IC_(50)、2IC_(50)作用于食管癌细胞Eca109、KYSE150,采用CCK-8法检测其抑制率;克隆形成实验检测克隆形成;划痕实验检测细胞迁移;Transwell小室实验检测细胞侵袭;免疫荧光染色检测细胞谷胱甘肽过氧化物酶4(GPX4)、上皮性钙黏附素(E-cadherin)、锌指转录因子(Slug)、神经性钙黏附蛋白(N-cadherin)表达;Western blot检测细胞E-cadherin、Slug、N-cadherin蛋白表达;ELISA检测细胞丙二醛(MDA)、超氧化物歧化酶(SOD)、还原型谷胱甘肽(GSH)、谷胱甘肽过氧化物酶(GPXS)活性;透射电镜观察细胞超微结构;普鲁士蓝染色观察细胞内铁颗粒分布。结果雄黄呈浓度和时间依赖性抑制Eca109、KYSE150细胞增殖,其IC_(50)分别为63.683μmol·L^(-1)、50.344μmol·L^(-1);与空白组相比,雄黄2IC_(50)组Eca109、KYSE150细胞迁移数、细胞侵袭数、N-cadherin、Slug、GPX4表达及MDA含量明显降低(P<0.05),E-cadherin表达和SOD、GSH、GPXS活性明显升高(P<0.05);且雄黄2IC_(50)组Eca109、KYSE150细胞内线粒体嵴变短变少甚至消失,发现大小各异的蓝染颗粒。结论雄黄可抑制Eca109、KYSE150细胞增殖、迁移和侵袭,诱导细胞铁死亡,可能是治疗食管癌潜在药物。
Aim To investigate the effects of realgar on the proliferation,invasion and ferroptosis of esophageal cancer cells.Methods Different concentrations of realgar(0,10,20,40,60,80,100μmol·L^(-1))or realgar 1/2IC_(50),IC_(50),2IC_(50)were applied to esophageal cancer cells Eca109 and KYSE150,and the inhibitory rate was detected by CCK-8 method;the clone formation ability was detected by clone formation assay;cell migration ability was detected by scratch assay;cell invasion was detected by Transwell assay;glutathione peroxidase 4(GPX4),epithelial cadherin(E-cadherin),zinc finger transcription factor(Slug),and neurocadherin(N-cadherin)expression were detected by immunofluorescence staining;E-cadherin,Slug,N-cadherin protein expression were detected by Western blot;intracellular malondialdehyde(MDA),superoxide dismutase(SOD),glutathione(GSH),glutathione peroxidase(GPXS)activity were assessed by enzyme linked immunosorbent assay(ELISA);cellular ultrastructure was observed by transmission electron microscopy;intracellular iron particle distribution was visualized by Prussian blue staining.Results Realgar inhibited the proliferation of Eca109 and KYSE150 cells in a concentration-and time-dependent manner,and the IC_(50)of realgar effect on Eca109 and KYSE150 cells was 63.683μmol·L^(-1)and 50.344μmol·L^(-1),respectively.Compared with control group,the migration number,cell invasion number,N-cadherin,Slug,GPX4 expression and MDA content of Eca109 and KYSE150 cells in the realgar 2IC_(50)group were significantly reduced(P<0.05),while E-cadherin expression and SOD,GSH and GPXS activities were significantly raised(P<0.05);and the mitochondrial cristae became shorter,fewer or even disappeared,and blue-stained particles of different sizes were found in Eca109 and KYSE150 cells in the realgar 2IC_(50)group.Conclusions Realgar can inhibit the proliferation of Eca109 and KYSE150 cells,inhibit cell migration and invasion,and induce cellular ferroptosis,which may be a potential drug for the treatment of esophageal cancer.
作者
陈发章
王君
徐海珍
郭占芳
杨如意
CHEN Fa-zhang;WANG Jun;XU Hai-zhen;GUO Zhan-fang;YANG Ru-yi(Medical College of Qinghai University,Affiliated Hospital of Qinghai University,Xining 810001,China;Dept of Integrated Traditional Chinese and Western Medicine,Affiliated Hospital of Qinghai University,Xining 810001,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2023年第1期108-115,共8页
Chinese Pharmacological Bulletin
基金
青海省科学技术厅课题(No2021-ZJ-730)。
关键词
食管癌
雄黄
铁死亡
细胞增殖
细胞迁移
细胞侵袭
esophageal cancer
realgar
ferroptosis
cell proliferation
cell migration
cell invasion
