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

Anticancer Res. 2017 Nov;37(11):6223-6234. doi: 10.21873/anticanres.12073.

Abstract

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 μM) and BITC (0, 0.5, 1 and 2 μM) 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 μM) and BITC (2 μM) decreased the levels of p-p38 following 24 and 48 h treatment. PEITC (1-2.5 μM) 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 μM) reduced the levels of p-ERK1/2 proteins following 48-h treatment but BITC (0.5-2 μM) 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-κB expression but BITC decreased NF-κB expression in the nucleus. Furthermore, NF-κB p65 binding to DNA was decreased following 2.5 μM PEITC treatment, but increased following treatment with 1-2 μM. However, 0.5-2 μM BITC treatment decreased the binding of NF-κB 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.

Keywords: BITC; MMP-2; NF-κB; PEITC; invasion; migration.

MeSH terms

  • Anticarcinogenic Agents / pharmacology*
  • Apoptosis / drug effects
  • Cell Movement / drug effects*
  • Cell Proliferation / drug effects
  • Gene Expression Regulation, Neoplastic / drug effects
  • Humans
  • In Vitro Techniques
  • Isothiocyanates / pharmacology*
  • MAP Kinase Signaling System / drug effects*
  • Melanoma / drug therapy
  • Melanoma / metabolism
  • Melanoma / pathology*
  • NF-kappa B / metabolism
  • Neoplasm Invasiveness
  • Signal Transduction / drug effects*
  • Tumor Cells, Cultured

Substances

  • Anticarcinogenic Agents
  • Isothiocyanates
  • NF-kappa B
  • phenethyl isothiocyanate
  • benzyl isothiocyanate