Effect of silencing ZEB1 gene on epithelial to mesenchymal transition in glioma U87 cells / 吉林大学学报(医学版)

ABSTRACT

Objective: To investigate the effects of silencing zinc finger E-box binding homeoboxl (ZEB1) gene on the expressions of mesenchymal markers and cell migration in the glioma U87 cells, and to clarify the effect of ZEB1 on the epithelial to mesenchymal transition (EMT) in the glioma cells. Methods: The constructed ZEB1 shRNA interfering plasmid and control plasmid (shCtrl) were transfected into the glioma U87 cells and the interfering effects were detected by Western blotting method. The glioma U87 cells were divided into control group (the glioma U87 cells were transfected with shCtrl), EMT group (EMT was induced by TGF-fil in the glioma U87 cells transfected with shCtrl) and ZEB1 silence group (EMT was induced by TGF-fil in the glioma U87 cells transfected with ZEB1 shRNAs plasmid). The protein expression levels of mesenchymal markers (N-cadherin, Vimentin), and matrix metalloproteinase-9 (MMP-9) in the glioma U87 cells were measured by Western blotting method. The scratch-healing assay was performed to examine the migration ability of glioma cells. Results: The Western blotting results showed that the expression levels of ZEB1 in the glioma U87 cells transfected with shZEBl # 1 and shZEBl # 2 were significantly lower than that in the cells transfected with shCtrl (P<0. 05 or P< 0. 01), and the inhibitory effect of shZEBl #2 on the ZEB1 expression was more obvious, indicating that ZEB1 was stably transfected into the U87 cells. Compared with control group, the expression levels of mesenchymal markers N-cadherin, Vimentin, and MMP-9 in EMT group were significantly increased (P<0. 05 or P<0. 01). Compared with EMT group, the expression levels of the above proteins in ZEB1 silencing group were markedly reduced (P< 0. 05 or P<0. 01). The cell migration rate in EMT group was obviously elevated compared with control group (P< 0. 01), and the cell migration rate of the glioma U87 cells in ZEB1 silence group was significantly lower than that in EMT group (P<0. 01). Conclusion: Silencing ZEB1 gene expression can inhibit the EMT in the glioma U87 cells and reduce the cell migration abilities, suggesting ZEB1 as an important therapeutic target of invasive glioma.