TGF-β1对HLEC发生EMT影响分析

doi:10.6040/j.issn.1673-3770.0.2016.477

山东大学耳鼻喉眼学报 ›› 2017, Vol. 31 ›› Issue (4): 76-79.doi: 10.6040/j.issn.1673-3770.0.2016.477

TGF-β1对HLEC发生EMT影响分析

王慧¹,温晏¹,闫丽¹,于晓明¹,宁宏²

1.山东省交通医院眼科, 山东济南 250031;
2.中国医科大学第四医院眼科, 辽宁沈阳 110000

收稿日期:2016-11-17 出版日期:2017-08-16 发布日期:2017-08-16
通讯作者: 于晓明. E-mail:qiaosong2443@163.com
基金资助:
辽宁省自然科学基金(2013021029)

The effect of transforming growth factor-β1 on epithelial-mesenchymal transition in human lens epithelial cells.

WANG Hui¹, WEN Yan¹, YAN Li¹, YU Xiaoming¹, NING Hong

1. Department of Ophthalmology, Shandong Provincial Traffic Hospital, Jinan 250031, Shandong, China;2. Department of Ophthalmology, The Fourth Hospital of China Medical University, Shenyang 110000, Liaoning, China

Received:2016-11-17 Online:2017-08-16 Published:2017-08-16

摘要/Abstract

摘要： 目的研究TGF-β1对人晶状体上皮细胞(HLEC)上皮间质转分化(EMT)的影响。方法根据处理不同,将细胞分为TGF-β1组、siRNA-TGF-β1组和空白对照组,培养24 h后观察细胞形态,Transwell法检测各组细胞迁移能力,实时荧光定量PCR检测各组细胞中TGF-β1、α-SMA、E-cadherin和vimentin基因表达,Western blotting法检测各组细胞中PI3K、Akt和p-Akt蛋白表达。结果 TGF-β1组迁移细胞数为(31.2±4.7)个,高于siRNA-TGF-β1组(14.6±5.3)个及空白对照组(3.9±1.4)个,且空白对照组高于siRNA-TGF-β1组;siRNA-TGF-β1组TGF-β1、α-SMA 及Vimentin的mRNA相对表达量均低于TGF-β1组及空白对照组,且空白对照组低于TGF-β1组;siRNA-TGF-β1组E-cadherin的mRNA相对表达量均高于TGF-β1组及空白对照组,且空白对照组高于TGF-β1组;siRNA-TGF-β1组细胞中PI3K蛋白及p-Akt蛋白相对表达量均低于TGF-β1组和空白对照组,且空白对照组低于TGF-β1组,siRNA-TGF-β1组Akt蛋白相对表达量高于TGF-β1处理组和空白对照组,且空白对照组高于TGF-β1组。结论 TGF-β1可诱导HLEC发生EMT,且可促进HLEC迁移,其机制可能与细胞中PI3K/Akt信号通路活化有关。

关键词: TGF-β1, 上皮-间质转化, PI3K/Akt信号通路, 人晶状体上皮细胞

Abstract: Objective To investigate the effect of transforming growth factor(TGF)-β1 on the epithelial-mesenchymal transition(EMT)in human lens epithelial cells(HLECs). Methods HLECs were cultured separately, according their treatment group. For this purpose, cells were divided into the TGF-β1 treatment group, siRNA-TGF-β1 group, and the control group. After culturing cells for 24 h in their respective treatment groups, cell morphologies were observed. Cell migration in each treatment group was detected using the transwell migration assay. The expression levels of TGF-β1, α-SMA, E-cadherin, and vimentin genes were detected using real-time polymerase chain reaction. The expression levels of PI3K, Akt, and p-Akt proteins were detected using Western blotting. Results In the TGF-β1 treatment group, 31.2±4.7 HLECs were found to have transformed. This number was significantly higher than that in the siRNA-TGF-β1(14.6±5.3)and control groups(3.9±1.4). Specifically, the number of cells transformed in the siRNA-TGF-β1 group was higher than that in the control group. Expression levels of TGF-β1 mRNA, α-SMA mRNA, and vimentin mRNA were the highest in the TGF-β1 treatment group, followed by lower levels in the siRNA-TGF-β1 group, and the lowest levels in the control group. In contrast, the expression levels of E-cadherin mRNA were highest in the siRNA-TGF-β1 group, followed by lower levels in the control group and the lowest levels in the TGF-β1 treatment group. These differences were statistically significant(P<0.05). The expression levels of PI3K and p-Akt proteins were the highest in the cells of the TGF-β1 treatment group, followed by lower levels in the control group. These levels were lowest in the siRNA-TGF-β1 group. The relative expression levels of Akt proteins were the highest in the siRNA-TGF-β1 group, followed by lower levels in the control group and the lowest levels in the TGF-β1 treatment group; these differences were statistically significant(P<0.05). Conclusion TGF-β1 could induce HLECs towards EMT, and thus promote HLEC migration. This mechanism could be related to the activation of PI3K/Akt signaling pathway.

Key words: TGF-β1, human lens epithelial cells, Epithelial-mesenchymal transition, PI3K/Akt signaling pathway

中图分类号:

R776.1

王慧,温晏,闫丽,于晓明,宁宏. TGF-β1对HLEC发生EMT影响分析[J]. 山东大学耳鼻喉眼学报, 2017, 31(4): 76-79.

WANG Hui, WEN Yan, YAN Li, YU Xiaoming, NING Hong. The effect of transforming growth factor-β1 on epithelial-mesenchymal transition in human lens epithelial cells.[J]. JOURNAL OF SHANDONG UNIVERSITY (OTOLARYNGOLOGY AND OPHTHALMOLOGY), 2017, 31(4): 76-79.

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TGF-β1对HLEC发生EMT影响分析

The effect of transforming growth factor-β1 on epithelial-mesenchymal transition in human lens epithelial cells.

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