摘要
目的探讨上皮⁃间质转化(epithelial⁃mesenchymal transition,EMT)过程中黑色素瘤细胞力学特性的变化规律及其与细胞迁移的内在联系。方法使用免疫荧光检测TGF⁃β1对EMT相关蛋白表达的影响;使用细胞划痕和Transwell小室检测细胞迁移和侵袭能力;借助原子力显微镜检测细胞刚度;通过流式细胞术检测骨架蛋白的动态变化。结果利用TGF⁃β1成功诱导A375细胞发生EMT。TGF⁃β1呈时间依赖性可增加波形蛋白vimentin表达,降低E⁃钙黏蛋白E⁃cadherin表达,并增强细胞迁移和侵袭能力。在EMT进程中,细胞的生物力学特性(细胞刚度)发生改变,细胞骨架G⁃actin/F⁃actin比值明显上升,细胞刚度明显降低。结论EMT进程中,黑色素瘤细胞骨架发生解聚,细胞刚度降低,从而增强细胞的迁移和侵袭能力。研究结果为癌症转移的临床治疗提供力学生物学新思路。
Objective To investigate the inner relationship of mechanical properties and migration during epithelial⁃mesenchymal transition(EMT)of melanoma A375 cells.Methods EMT was induced by TGF⁃β1 in epithelial⁃like melanoma A375 cells,and further morphological observation and analysis of A375 cells were performed.The effect of TGF⁃β1 on EMT⁃related protein expression was detected by immunofluorescence.Cell scratch and transwell assay were used to detect cell migration and invasion.The cell stiffness was measured by atomic force microscopy(AFM).The cytoskeletal proteins of G⁃actin and F⁃actin were determined by flow cytometry.Results Melanoma A375 cells were successfully induced by TGF⁃β1 stimulation to establish the EMT phenotype.TGF⁃β1 increased the expression of mesenchymal marker of vimentin and decreased the expression of epithelial marker of E⁃cadherin in a time⁃dependent manner,which further enhanced cell migration and invasion.TGF⁃β1⁃induced EMT also altered the biomechanical properties of melanoma A375 cells,which showed a significant increase in cytoskeletal G⁃actin/F⁃actin ration and an obvious decrease in cell stiffness.Conclusions The cytoskeleton of melanoma A375 cells undergoes depolymerization and cell stiffness decreases in response to TGF⁃β1⁃induced EMT,which enhances the ability of cell migration and invasion.These research findings provide new strategy in mechanobiology for clinical treatment of cancer metastasis.
作者
陈瑜
姜晓敏
李亭亭
李顺
秦翔
刘贻尧
CHEN Yu;JIANG Xiaomin;LI Tingting;LI Shun;QIN Xiang;LIU Yiyao(School of Life Science and Technology,University of Electronic Science and Technology of China,Chengdu 610054,China)
出处
《医用生物力学》
CAS
CSCD
北大核心
2022年第5期874-880,共7页
Journal of Medical Biomechanics
基金
国家自然科学基金项目(U19A2006,12132004)。
关键词
上皮间质转化
细胞骨架
细胞刚度
细胞迁移
epithelial⁃mesenchymal transition(EMT)
cytoskeleton
cell stiffness
cell migration
作者简介
通信作者:刘贻尧,教授,博士生导师,E⁃mail:liuyiyao@uestc.edu.cn。
