TGF-β1对胶质瘤U87细胞上皮-间质转化的诱导作用  

Induction of TGF-β1 on epithelial to mesenchymal transition in glioma U87 cells

作　　者：宋扬[1] 赵丽艳[2] 陈勇[1] 吕晓艳[2] 崔佳乐[3] 李蕴潜[1] 赵刚[1] 

机构地区：[1]吉林大学第一医院神经外科,吉林长春130021 [2]吉林大学第二医院检验科,吉林长春130041 [3]吉林大学基础医学院组织学与胚胎学系,吉林长春130021

出　　处：《吉林大学学报（医学版）》2016年第6期1087-1091,I0013,共6页Journal of Jilin University：Medicine Edition

基　　金：吉林省科技厅科技发展计划项目资助课题(20150414024GH)

摘　　要：目的:探讨转化生长因子β1(TGF-β1)诱导胶质瘤U87细胞发生上皮-间质转化(EMT)的作用,为阐明胶质瘤侵袭性生长的机制提供依据。方法:以不同浓度TGF-β1体外处理胶质瘤U87细胞,分为2.5×10^(-6)、5.0×10^(-6)、10.0×10^(-6)和20.0×10^(-6)g·L^(-1) TGF-β1组,以0×10^(-6)g·L^(-1) TGF-β1组作为空白对照组。Western blotting法检测各组U87细胞中上皮标志物(上皮钙黏蛋白)和间质标志物(神经钙黏蛋白和波形蛋白)以及转录因子(Snail、Slug和Zeb1蛋白)的相对表达水平;倒置显微镜下观察U87细胞形态表现;划痕愈合实验检测U87细胞相对迁移距离。结果:随着TGF-β1浓度的升高,胶质瘤U87细胞中上皮钙黏蛋白相对表达水平逐渐降低,2.5×10^(-6)、5.0×10^(-6)、10.0×10^(-6)和20.0×10^(-6) g·L^(-1) TGF-β1组U87细胞中上皮钙黏蛋白相对表达水平低于空白对照组(P<0.05或P<0.01);神经钙黏蛋白和波形蛋白相对表达水平则随着TGF-β1浓度的升高逐渐升高,神经钙黏蛋白相对表达水平分别比空白对照组升高11.5%(P>0.05)、39.4%(P>0.05)、51.0%(P<0.05)和76.9%(P<0.05);10.0×10^(-6)和20.0×10^(-6)g·L^(-1) TGF-β1组U87细胞中波形蛋白相对表达水平均高于空白对照组(P<0.01)。5.0×10^(-6)、10.0×10^(-6)和20.0×10^(-6) g·L^(-1) TGF-β1组U87细胞中Snail、Slug和Zeb1蛋白的相对表达水平高于空白对照组(P<0.05或P<0.01)。与空白对照组比较,10.0×10^(-6) g·L^(-1) TGF-β1组U87细胞呈拉伸状和更为狭长,细胞迁移能力明显增强。结论:一定浓度范围内,TGF-β1能以浓度依赖方式诱导胶质瘤U87细胞发生EMT。Objective：To investigate the induction of transforming growth factor beta 1（TGF-β1）on epithelial to mesenchymal transition（EMT）in glioma U87 cells,and to provide evidence to clarify the mechanism of glioma invasion growth.Methods：The glioma U87 cells were treated with different concentrations of TGF-β1 in vitro and divided into 2.5×10^-6,5.0×10^-6,10.0×10^-6,and 20.0 ×10^-6g·L^（-1） TGF-β1groups,and 0×10^-6g·L^（-1）TGF-β1treatment was regarded as blank control group.The relative expression levels of epithelial marker（E-cadherin）,mesenchymal markers（N-cadherin,Vimentin）,and transcription factors（Snail,Slug,and Zeb1proein）in the U87 cells in various groups were detected by Western blotting method;the morphology of U87 cells was observed by inverted microscope;the relative wound space of U87 cells was examined by scratch healing assay.Results：With the increasing of concentrations of TGF-β1,the relative expression levels of E-cadherin in glioma U87 cells in 2.5×10^-6,5.0×10^-6,10.0×10^-6,and 20.0×10^-6 g·L^（-1） groups were gradually decreased compared with blank control group（P〈0.05 or P〈0.01）.The relative expression levels of N-cadherin and Vimentin showed an increasing trend,and the relative expression levels of N-cadherin were increased by 11.5%（P〈0.05）,39.4%（P〈0.05）,51.0%（P〈0.05）,and 76.9%（P〈0.05）in 2.5×10^-6,5.0×10^-6,10.0×10^-6,and20.0×10^-6 g·L^（-1） TGF-β1groups compared with blank control groups.The relative expression levels of Vimentin in the U87 cells in TGF-β1 10×10^-6 and 20×10^-6g·L^（-1） groups were higher than those in blank control group（P〈0.01）.The relative expression levels of Snail,Slug and Zeb1 proteins in the U87 cells in 5.0×10^-6,10.0×10^-6,and 20.0×10^-6 g·L^（-1） groups were higher than those in blank control group（P〈0.05 or P〈0.01）.Compared with blank control group,the morphology of U87 cells in 10×10^-6g·L^（-1） TGF-β1group became more stretching,long and narrow,

关 键 词：胶质瘤细胞 转化生长因子Β1 上皮-间质转化 钙黏蛋白 转录因子 

分 类 号：R739.4[医药卫生—肿瘤]