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 si RNA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。
Objective To observe the role ofβ-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 ofβ-catenin siRNA or the S33Y-β-catenin cDNA expression vector for descending or over expression ofβ-catenin.Western Blot was applied to detect the expression ofβ-catenin,Wnt5a,E-cad-herin andα-SMA proteins in each group.Results After the addition of TGF-β2,the expression of Wnt5a protein andβ-catenin protein in SRA01/04 cells was significantly up-regulated;the relative expression of E-cadherin protein decreased,and the rel-ative expression ofα-SMA protein increased.After transfection withβ-catenin-siRNA,the expression ofβ-catenin protein was significantly down-regulated,while there was no significant change in the expression of Wnt5a protein;After stimulation withβ-catenin-siRNA+TGF-β2,the expression of E-cadherin protein was up-regulated,whereas the expression ofβ-catenin,Wnt5a and a-SMA proteins were down-regulated.After transfection of S33Y-β-catenin cDNA,the expression ofβ-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 andβ-catenin in normal lens epithe-lial cells,and induced EMT;overexpression or degradation ofβ-catenin can promote or decrease the expression of TGF-β2-in-duced Wnt5a,and participate in TGF-β2-induced EMT.
作者
王景瑶
包秀丽
WANG Jingyao;BAO Xiui(College of Graduate,Inner Mongolia Medical University,Hohhot 010059,China;Department of Ophthalmology,The Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010050,China)
出处
《内蒙古医科大学学报》
2024年第2期117-121,共5页
Journal of Inner Mongolia Medical University
基金
内蒙古自治区自然科学基金项目(2019MS08116)。
作者简介
第一作者:王景瑶(1996—),女,2021级在读硕士研究生。E-mail:754212491@qq.com;通信作者:包秀丽,女,医学博士,主任医师,硕士研究生导师。研究方向:白内障的临床和基础研究、小儿眼科。E-mail:ophbaxili@hotmail.com。
