半乳糖凝集素3调控转化生长因子-β诱导的A549细胞上皮间质转化

摘要

目的 探讨半乳糖凝集素3表达对转化生长因子-β(TGF-β)诱导A549细胞上皮间质转化(EMT)的影响.方法 在A549细胞系中过表达半乳糖凝集素3,TGF-β (20 nmol/L)诱导肺癌A549细胞EMT.蛋白质印迹法检测半乳糖凝集素3、钙黏附蛋白E和波形蛋白等相关蛋白的表达.免疫荧光检测钙黏附蛋白E等蛋白的表达以及细胞形态的变化,细胞计数试剂盒-8检测细胞增殖,划痕愈合实验检测细胞迁移性的变化,细胞侵袭性实验检测细胞侵袭性的变化.结果 在过表达半乳糖凝集素3的A549细胞系中,EMT的相关蛋白如钙黏附蛋白E、波形蛋白表达水平无变化,细胞增殖、侵袭、迁移能力无变化;在TGF-β诱导肺癌A549细胞EMT的模型中,半乳糖凝集素3过表达组EMT相关蛋白钙黏附蛋白E表达下调、波形蛋白表达上调.细胞增殖能力无变化,细胞侵袭、迁移能力增强.结论 半乳糖凝集素3可以促进TGF-β诱导的A549细胞上皮间质转化.%Objective To explore the role of Galectin3 in transforming growth factor-β (TGF-β)-induced epithelial-mesenchymal transition (EMT) in A549 cells.Methods Galectin3 was over-expressed in an A549 cell line.EMT was induced in lung cancer A549 cells by adding TGF-β.The expressions of Galectin3,Ecadherin,and vimentin were determined by Western blot.The protein expression of E-cadherin and the morphological changes of the cells were detected by immunofluorescence.Cellular proliferation was analyzed with cell counting kit-8,and the cellular migration and invasion was measured by scratches healing and Transwell assay,respectively.Results When only Galectin3 was over-expressed in A549 cell line,the expression levels of EMT-related proteins such as E-cadherin and vimentin were not changed,and the abilities of cellular proliferation,invasion,and migration were not changed either.When the EMT was induced by TGF-β in A549 cells,the E-cadherin expression was down-regulated and the vimentin expression was up-regulated in A549 cells with Galectin3 overexpression.There was no significant change in cellular proliferation,whereas the abilities of cellular invasion and migration were enhanced.Conclusion The TGF-β-induced EMT in A549 cells can be enhanced by Galectin3.