MgCl2 and ZnCl2 promote human umbilical vein endothelial cell migration and invasion and stimulate epithelial-mesenchymal transition via the Wnt/β-catenin pathway.

Abstract

Previous studies have demonstrated that magnesium and zinc ions promote the migration and epithelial-mesenchymal transition (EMT) of cancer/endothelial cells. However, the impact of MgCl2 and ZnCl2 on the migration, invasion and EMT of human umbilical vein endothelial cells (HUVECs) and the involved mechanisms remain unclear. In the present study, HUVECs were incubated with various doses of MgCl2 and ZnCl2. The optimum concentrations of MgCl2 and ZnCl2 were selected by MTT assay. The migration and invasion capabilities of HUVECs were analyzed by Transwell assays. Subsequently, the expression of matrix metalloproteinase (MMP)-2 and MMP-9 mRNA and protein were determined by reverse transcription-quantitative polymerase chain reaction, western blotting and ELISA. MMP-2 and MMP-9 activities were measured by gelatin zymography. Immunofluorescence staining was performed to investigate cytoskeletal dynamics using Acti-stain™ 488 Fluorescent Phalloidin. Subsequently, the expression of EMT-related markers at the mRNA and protein levels and the activation of Wnt/β-catenin signaling were analyzed. The results identified increases in MMP-2 and MMP-9 expression and activity, indicating that MgCl2 and ZnCl2 promoted HUVEC migration and invasion. In addition, MgCl2 and ZnCl2 treatment induced cytoskeleton remodeling and stimulated EMT via activation of the Wnt/β-catenin signaling pathway, characterized by a decrease in E-cadherin and increases in N-cadherin, vimentin and Snail. These results suggest that MgCl2 and ZnCl2 may enhance the migration and invasion capabilities of HUVECs and promote EMT through the Wnt/β-catenin pathway.