TUBA1A regulates epithelial-mesenchymal transition in glioma cells by modulating Smad-dependent signaling

WANG Shuren; SHAO Weiwei; LAN Jianyun

doi:10.3969/j.issn.1674-4136.2024.06.010

Abstract：Objective To explore the effect and mechanism of TUBA1A expression on epithelial-mesenchymal transition(EMT) of glioma cells.Methods The Glivis database was used to analyze the expression of TUBA1A in glioma tissues,and Western blot experiments were used to verify the differences between Chinese glioma tissues and normal brain tissues at all levels.Exogenous administration of TGF-β1 induced EMT in glioma U87 and U251cells,and was verified by detecting changes in EMT-related proteins ZO-1,N-cadherin and Vimentin.After U87 andU251cells were transfected with TUBA1A siRNA,changes in cell invasion and migration were analyzed by Transwell,and changes in MMP-2,MMP-9,ZO-1,N-cadherin and Vimentin were verified by Western blot.Further,the expression of Smad pathway-related proteins after TUBA1A siRNA transfection was detected.Results Glivis database analysis showed that TUBA1A is overexpressed in glioblastoma ( GBM),and Western blot results also showed that the expression of TUBA1A in Chinese high-grade glioma tissues was significantly higher than that in low-grade glioma tissues and normal brain tissue.After exogenous TGF-β1 intervention,the EMT-related protein ZO-1 was significantly reduced,while the expressions of N-cadherin and Vimentin were significantly increased.After U87 andU251cells were transfected with TUBA1A siRNA,the cell invasion and migration abilities were significantly reduced,the expression of MMP-2,MMP-9,N-cadherin and Vimentin proteins was significantly reduced,and the expression of ZO-1 was significantly increased.It was further found that the phosphorylation levels of Smad pathway-related proteins Smad2 and Smad3 were significantly reduced.Conclusions TUBA1A is closely related to the EMT transformation process of glioma cells induced by TGF-β1,and enhances the epithelial-mesenchymal transition ability of cells through activation of the Smad pathway.