TGF-β1对胶质瘤U87细胞上皮-间质转化的诱导作用

doi:10.13481/j.1671-587x.20160609

吉林大学学报(医学版) ›› 2016, Vol. 42 ›› Issue (06): 1087-1091.doi: 10.13481/j.1671-587x.20160609

TGF-β1对胶质瘤U87细胞上皮-间质转化的诱导作用

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

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

收稿日期:2016-07-08 出版日期:2016-11-28 发布日期:2016-12-02
通讯作者: 赵丽艳,副教授,硕士研究生导师(Tel:0431-88796607,E-mail:970201043@qq.com);赵刚,教授,博士研究生导师(Tel:0431-88782331,E-mail:zhaogang@jdnwk.com) E-mail:970201043@qq.com;zhaogang@jdnwk.com
作者简介:宋扬(1990-),男,吉林省长春市人,在读医学博士,主要从事胶质瘤基础和临床方面的研究。
基金资助:
吉林省科技厅科技发展计划项目资助课题（20150414024GH）

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

SONG Yang¹, ZHAO Liyan², CHEN Yong¹, LYU Xiaoyan², CUI Jiayue³, LI Yunqian¹, ZHAO Gang¹

1. Department of Neurosurgery, First Hospital, Jilin University, Changchun 130021, China;
2. Department ofClinical of Medical Laboratory, Second Hospital, Jilin University, Changchun 130041, China;
3. Department of Histology and Embryology, School of Basic Medical Sciences, Jilin University, Changchun 130021, China

Received:2016-07-08 Online:2016-11-28 Published:2016-12-02

摘要/Abstract

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^-6g·L^-1 TGF-β1组，以0×10^-6g·L^-1TGF-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^-6g·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^-6g·L^-1TGF-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^-6g·L^-1TGF-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^-1TGF-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。

关键词: 1, 胶质瘤细胞, 上皮-间质转化, 钙黏蛋白, 转录因子, 转化生长因子&beta

Abstract:

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-β1 groups,and 0×10^-6g·L^-1 TGF-β1 treatment 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 Zeb1 proein) 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, and 20.0×10^-6 g·L^-1 TGF-β1 groups 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^-6g·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^-1TGF-β1 group became more stretching,long and narrow, and the cell migration ability was also significantly enhanced.Conclusion: Within a certain concentration range,TGF-β1 could induce EMT in glioma U87 cells in a concentration-dependent manner.

Key words: transforming growth factor beta 1, epithelial to mesenchymal transition, cadherin, transcription factor, glioma cell

中图分类号:

R739.4

宋扬, 赵丽艳, 陈勇, 吕晓艳, 崔佳乐, 李蕴潜, 赵刚. TGF-β1对胶质瘤U87细胞上皮-间质转化的诱导作用[J]. 吉林大学学报(医学版), 2016, 42(06): 1087-1091.

SONG Yang, ZHAO Liyan, CHEN Yong, LYU Xiaoyan, CUI Jiayue, LI Yunqian, ZHAO Gang. Induction of TGF-β1 on epithelial to mesenchymal transition in glioma U87 cells[J]. Journal of Jilin University Medicine Edition, 2016, 42(06): 1087-1091.

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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

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