氧化克班宁促进人乳腺癌MCF-7细胞微管解聚的机制研究  

Study on the mechanisms of promoting microtubule depolymerization by oxocrebanine in human breast cancer MCF⁃7 Cells

作　　者：于蕾[1] 闫彩凤 宋辉[1] 郎朗 于晶涵 马瑗晗 王正文 YU Lei;YAN Caifeng;SONG Hui;LANG Lang;YU Jinghan;MA Yuanhan;WANG Zhengwen(College of Pharmacy/Drug Engineering Technology Research Center,Harbin University of Commerce,Harbin 150076,China;Hepatobiliary and Pancreatic Surgery,Hainan Cancer Hospital,Haikou 570000,China)

机构地区：[1]哈尔滨商业大学药学院/药物工程技术研究中心,黑龙江哈尔滨150076 [2]海南省肿瘤医院肝胆胰外科,海南海口570000

出　　处：《海南医科大学学报》2025年第4期264-269,共6页Journal of Hainan Medical University

基　　金：海南省自然科学基金项目(820RC776);海南省卫生健康科技创新联合项目(WSJK2024MS162);2021年哈尔滨商业大学教师“创新”项目支持计划项目(LH2020H070);黑龙江省中医药科研课题项目(ZHY2024-098)。

摘　　要：目的:探究氧化克班宁通过Rho/ROCK信号通路,促进人乳腺癌MCF-7细胞微管解聚的作用机制。方法:首先采用免疫荧光法,研究氧化克班宁对MCF-7细胞骨架关键蛋白F-actin的影响;其次,采用Western Blot法检测氧化克班宁作用MCF-7细胞后,F-actin的蛋白表达水平、Rho/ROCK信号通路关键性蛋白Rho A、ROCK1和ROCK2的蛋白表达水平,以及经氧化克班宁和抑制剂GSK429286A联用处理后,MCF-7细胞内Rho/ROCK下游蛋白MYPT和LIMK的磷酸化水平。结果:氧化克班宁作用MCF-7细胞48 h后,与空白对照组相比,F-actin蛋白荧光强度逐渐降低,F-actin蛋白表达水平显著降低(P<0.01),Rho A、ROCK1和ROCK2蛋白表达水平显著降低(P<0.01);氧化克班宁与抑制剂GSK429286A联用,MCF-7细胞内pMYPT1及pLIMK1蛋白表达水平均显著降低(P<0.01)。结论:氧化克班宁可破坏细胞骨架,干预Rho/ROCK通路,调控其下游蛋白MYPT和LIMK磷酸化水平,促进微管解聚,抑制人乳腺癌MCF-7细胞生长。Objective:This paper explores the mechanism of oxocrebanine in promoting microtubule depolymerization of human breast cancer MCF‑7 cells through the Rho/ROCK signaling pathway.Methods:Firstly,immunofluorescence was used to study the effect of oxocrebanine on F‑actin,a key protein in the skeleton of MCF‑7 cells.Secondly,the Western Blot method was used to detect the protein expression levels of F‑actin and Rho A,ROCK1 and ROCK2,key proteins of the Rho/ROCK signaling pathway.At last,the phosphorylation level of Rho/ROCK downstream proteins MYPT and LIMNK were detected by Western Blot method while MCF‑7 cells were treated with GSK429286A and oxocrebanine.Results:After MCF‑7 cells were treated with oxocrebanine for 48 h,the fluorescence intensity of F‑actin protein gradually decreased,the expression level of F‑actin protein was also significantly reduced(P<0.01),and the expression levels of Rho A,ROCK1 and ROCK2 protein were significantly reduced(P<0.01).After MCF‑7 cells were treated with GSK429286A and oxocrebanine,the phosphorylation levels of MYPT and LIMK,Rho/ROCK downstream proteins,were reduced in a dose‑dependent manner(P<0.01).Conclusion:Oxocrebanine can destroy the cytoskeleton,interfere in the Rho/ROCK pathway,regulate the phosphorylation levels of its downstream proteins MYPT and LIMK,promote microtubule depolymerization and inhibit human breast cancer MCF‑7 cells growth.

关 键 词：海南地不容 氧化克班宁 MCF-7细胞 微管 Rho/ROCK通路 

分 类 号：R285[医药卫生—中药学]