氯氧喹通过下调Rho/Rho激酶信号通路抑制乳腺癌细胞转移  

Chloroxoquinoline inhibits invasion in breast cancer via down-regulating Rho/Rho kinase signaling pathway

作　　者：刘婧雯 杨兴莲 沈凯莉 曾玲晖[1] 孙燕[2] LIU Jingwen;YANG Xinglian;SHEN Kaili;ZENG Linghui;SUN Yan(School of Medicine,Zhejiang University City College,Hangzhou 310015,China;Department of Oncology,Zhejiang Cancer Hospital,Hangzhou 310022,China)

机构地区：[1]浙江大学城市学院医学院,浙江杭州310015 [2]浙江省肿瘤医院综合肿瘤内科,浙江杭州310022

出　　处：《浙江大学学报（医学版）》2019年第6期631-637,共7页Journal of Zhejiang University(Medical Sciences)

基　　金：浙江省医药卫生科技计划(2015RCB004);杭州市留学回国人员资助项目(2018)

摘　　要：目的:通过观察氯氧喹对乳腺癌细胞系细胞骨架的聚合状态及其对Rho/Rho激酶信号通路相关靶点蛋白磷酸化的作用,明确氯氧喹治疗乳腺癌的作用机制。方法:以不同浓度氯氧喹处理乳腺癌Bcap37和MDA-MB--453细胞系,划痕试验检测细胞迁移,Transwell试验检测细胞侵袭,罗丹明标记鬼笔环肽染色法观察F肌动蛋白的聚解状态,免疫荧光法观察α微管蛋白的表达,蛋白质印迹法检测Rho/Rho激酶信号通路关键蛋白磷酸化水平。结果:氯氧喹可剂量依赖性地抑制细胞迁移和侵袭;Bcap37和MDA-MB-453细胞经氯氧喹处理后其细胞形态发生改变,细胞体积减少;F肌动蛋白和α微管蛋白染色形态不规则,荧光染色聚集成团,且呈现剂量依赖性;氯氧喹可下调Rho/Rho激酶信号通路中肌动蛋白解聚因子Cofilin、磷酸化蛋白激酶(Limk)、Rho关联卷曲蛋白激酶2(Rock2)磷酸化表达(均P<0.01)。结论:氯氧喹可抑制乳腺癌细胞骨架聚合从而抑制细胞迁移,该作用可能与抑制Rho/Rho激酶信号通路相关。Objective:To investigate the effect of chloroxoquinoline on cytoskeleton of breast cancer cells and its relation with Rho/Rho kinase signaling pathway.Methods:Breast cancer Bcap37 and MDA-MB-453 cells were treated with different concentrations of chloroxoquinoline.Wound healing and Transwell assay were conducted to detect cell migration and invasion,respectively.Rhodamine-phalloidin staining and immunofluorescent staining were used to observe the polymerization state of F-actin and the expression ofα-Tublin in breast cancer cells,respectively.Western blot was used to detect the phosphorylation level of key protein in Rho/Rho kinase signaling pathway.Results:Compared with the control group,chloroxoquinoline treatment induced dose-dependent decrease in cell migration and invasion,and Bcap37 and MDA-MB-453 cells treated with chloroxoquinoline showed dose-dependent changes in cell morphology and decrease in cell body.The staining of F-actin andα-Tublin was irregular and clustered.Furthermore,treatment of chloroxoquinoline down-regulated the phosphorylation of the Rho/Rho kinase signaling proteins Cofilin,Limk and Rock2(all P<0.01).Conclusions:Chloroxoquinoline inhibits the cytoskeleton in breast cancer Bcap37 and MDA-MB-453 cells and inhibits cell migration.This effect may be associated with down-regulation of Rho/Rho kinase signaling pathway.

关 键 词：乳腺肿瘤/病理学 细胞骨架 rho相关激酶类/拮抗剂和抑制剂 信号传导/药物作用 肌动蛋白类 微管蛋白 抗肿瘤药/药理学 

分 类 号：R737.9[医药卫生—肿瘤]