Objective To investigate the inner relationship of mechanical properties and migration during epithelial-mesenchymal transition (EMT) of melanoma A375 cells. Methods EMT was induced by TGF-β1 in epithelial-like melanoma A375 cells, and further morphological observation and analysis of A375 cells were performed. The effect of TGF-β1 on EMT-related protein expression was detected by immunofluorescence. Cell scratch and transwell assay were used to detect cell migration and invasion. The cell stiffness was measured by atomic force microscopy (AFM). The cytoskeletal proteins of G-actin and F-actin were determined by flow cytometry. Results Melanoma A375 cells were successfully induced by TGF-β1 stimulation to establish the EMT phenotype. TGF-β1 increased the expression of mesenchymal marker of vimentin and decreased the expression of epithelial marker of E-cadherin in a timedependent manner, which further enhanced cell migration and invasion. TGF-β1-induced EMT also altered the biomechanical properties of melanoma A375 cells, which showed a significant increase in cytoskeletal G-actin/F-actin ration and an obvious decrease in cell stiffness. Conclusions The cytoskeleton of melanoma A375 cells undergoes depolymerization and cell stiffness decreases in response to TGF-β1-induced EMT, which enhances the ability of cell migration and invasion. These research findings provide new strategy in mechanobiology for clinical treatment of cancer metastasis.