β-连环素调控Wnt5a参与人晶状体上皮细胞转分化
β-CATENIN REGULATES Wnt5a AND PARTICIPATES IN EPITHELIAL MESENCHYMAL TRANSDIFFERENTIATION OF HUMAN LENS EPITHELIAL CELLS
王景瑶 1包秀丽2
作者信息
- 1. 内蒙古医科大学研究生院,内蒙古 呼和浩特 010059
- 2. 内蒙古医科大学附属医院眼科,内蒙古 呼和浩特 010050
- 折叠
摘要
目的 观察在转化生长因子-β2(transform growth factor-β2,TGF-β2)调控Wnt5a表达中,β-连环蛋白(β-catenin)的作用及其对TGF-β2诱导的上皮间充质转化(epithelial mesenchymal transition,EMT)的调控作用.方法 以体外培养的人晶状体上皮细胞(SRA01/04细胞)为研究对象,加入外源性10 ng/mL TGF-β2来诱导细胞EMT.在加入前48 h,采用脂质体介导转染技术瞬时转染β-catenin siRNA或S33Y-β-catenin cDNA表达载体,作为β-catenin降表达或过表达的干预处理.应用Western Blot印迹法检测各组中β-catenin、Wnt5a、E-cadherin、α-SMA蛋白的表达情况.结果 加入TGF-β2后,SRA01/04细胞Wnt5a蛋白、β-catenin蛋白的表达明显上调;E-cadherin蛋白相对表达量下降,α-SMA蛋白的相对表达量增加.转染β-catenin-siRNA后,β-catenin蛋白的表达明显下调,而Wnt5a蛋白表达无明显变化;β-catenin-siRNA+TGF-β2刺激后,E-cadherin蛋白表达上调,而β-catenin、Wnt5a和a-SMA蛋白的表达下调.转染S33Y-β-catenin cDNA后,β-catenin蛋白表达明显升高,而Wnt5a蛋白表达无明显变化.S33Y-β-catenin+TGF-β2刺激后,α-SMA蛋白的表达明显上调,E-cadherin蛋白的表达下调.结论 TGF-β2诱导正常人晶状体上皮细胞过表达Wnt5a和β-catenin蛋白,并诱导EMT;β-catenin的过表达或降表达能促进或降低TGF-β2诱导Wnt5a的表达,参与TGF-β2诱导的EMT.
Abstract
Objective To observe the role of β-catenin in the regulation of Wnt5a expression by transform growth factor-β2(TGF-β2)and its effect on TGF-β2-induced epithelial mesenchymal transition(EMT).Methods Take human lens epithelial cells(SRA01/04 cells)which were cultured in vitro as research object,added exogenous 10 ng/mL TGF-β2 to induce cellular EMT.48 hours prior to the addition of TGF-β2,liposome-mediated transfection technology was employed as an intervention treatment.This process involved the transient transfection of β-catenin siRNA or the S33Y-β-catenin cDNA expression vector for descending or over expression of β-catenin.Western Blot was applied to detect the expression of β-catenin,Wnt5a,E-cad-herin and α-SMA proteins in each group.Results After the addition of TGF-β2,the expression of Wnt5a protein and β-catenin protein in SRA01/04 cells was significantly up-regulated;the relative expression of E-cadherin protein decreased,and the rel-ative expression of α-SMA protein increased.After transfection with β-catenin-siRNA,the expression of β-catenin protein was significantly down-regulated,while there was no significant change in the expression of Wnt5a protein;After stimulation with β-catenin-siRNA+TGF-β 2,the expression of E-cadherin protein was up-regulated,whereas the expression of β-catenin,Wnt5a and a-SMA proteins were down-regulated.After transfection of S33Y-β-catenin cDNA,the expression of β-catenin protein was significantly elevated,whereas there was no significant change in the expression of Wnt5a protein.α-SMA protein expression was significantly up-regulated and E-cadherin protein expression was down-regulated after stimulation with S33Y-β-catenin+TGF-β2.Conclusions TGF-β2 induced overexpression of Wnt5a and β-catenin in normal lens epithe-lial cells,and induced EMT;overexpression or degradation of β-catenin can promote or decrease the expression of TGF-β2-in-duced Wnt5a,and participate in TGF-β2-induced EMT.
关键词
TGF-β/Wnt5a/β-catenin/EMT/后囊膜混浊Key words
TGF-β/Wnt5a/β-catenin/EMT/PCO引用本文复制引用
基金项目
内蒙古自治区自然科学基金项目(2019MS08116)
出版年
2024
NETL
NSTL
万方数据