Phenethyl Isothiocyanate (PEITC) and Benzyl Isothiocyanate (BITC) Inhibit Human Melanoma A375. S2 Cell Migration and Invasion by Affecting MAPK Signaling Pathway In Vitro

Background/Aim: Numerous evidence has shown that PEITC and BITC inhibit cancer cell migration and invasion. In this study, we investigated the anti-metastatic mechanisms of PEITC and BITC in human melanoma cancer A375.S2 cells in vitro. Materials and Methods: We used a cell viability assay, an in-vitro scratch wound healing assay, a transwell assay for cell migration and invasion, a gelatin zymography assay, western blotting and EMSA to examine the anti-metastatic mechanisms of PEITC and BITC in A375.S2 cells. Results: Sublethal concentrations of PEITC (0, 1, 2 and 2.5 mu M) and BITC (0, 0.5, 1 and 2 mu M) inhibited mobility, migration and invasion of A375.S2 cells that were assayed by wound healing and Transwell filter. PEITC and BITC inhibited MMP-2 activity in A375.S2 cells, as assessed by gelatin zymography assay. Results from western blotting indicated that PEITC (2.5 mu M) and BITC (2 mu M) decreased the levels of p-p38 following 24 and 48 h treatment. PEITC (1-2.5 mu M) reduced the levels of p-JNK1/2 proteins following 48-h treatment but BITC increased p-JNK1/2 levels following 24-h treatment. PEITC (2.5 mu M) reduced the levels of p-ERK1/2 proteins following 48-h treatment but BITC (0.5-2 mu M) increased p-ERK1/2 levels following 24- and 48-h treatment. PEITC and BITC affect cell migration and invasion of A375.S2 cells via MAPK pathway. PEITC and BITC inhibited MMP-2 activity. PEITC increased NF-kappa B expression but BITC decreased NF-kappa B expression in the nucleus. Furthermore, NF-kappa B p65 binding to DNA was decreased following 2.5 mu M PEITC treatment, but increased following treatment with 1-2 mu M. However, 0.5-2 mu M BITC treatment decreased the binding of NF-kappa B to DNA in A375.S2 cells, as assessed by electrophoretic mobility shift (EMSA) assay. Conclusion: Based on these observations, we suggest that PEITC and BITC can be used as anti-metastastic agents of human melanoma cells in the future.