Phenethyl isothiocyanate triggers apoptosis in human malignant melanoma A375.S2 cells through reactive oxygen species and the mitochondria-dependent pathways

Hum Exp Toxicol. 2014 Mar;33(3):270-83. doi: 10.1177/0960327113491508. Epub 2013 Jun 11.


We have reported previously that phenethyl isothiocyanate (PEITC) induces apoptosis in human osteosarcoma U-2 OS cells. Cytotoxic activity of PEITC towards other cancer cells such as human malignant melanoma and skin cancer cells has not been reported. In this study, the anticancer activity of PEITC towards human malignant melanoma cancer A375.S2 cells was investigated. To determine the mechanisms of PEITC inhibition of cell growth, the following end points were determined in A375.S2 cells: cell morphological changes, cell cycle arrest, DNA damage and fragmentation assays and morphological assessment of nuclear change, reactive oxygen species (ROS) and Ca(2+) generations, mitochondrial membrane potential disruption, and nitric oxide and 10-N-nonyl acridine orange productions, expression and activation of caspase-3 and -9, B-cell lymphoma 2 (Bcl-2)-associated X protein (Bax), Bcl-2, poly (adenosine diphosphate-ribose) polymerase, and cytochrome c release, apoptosis-inducing factor and endonuclease G. PEITC induced morphological changes in time- and dose-dependent manner. PEITC induced G2/M phase arrest and induced apoptosis via endoplasmic reticulum stress-mediated mitochondria-dependent pathway. Western blot analysis showed that PEITC promoted Bax expression and inhibited Bcl-2 expression associated with the disintegration of the outer mitochondrial membrane causing cytochrome c release, and activation of caspase-9 and -3 cascade leading to apoptosis. We conclude that PEITC-triggered apoptotic death in A375.S2 cells occurs through ROS-mediated mitochondria-dependent pathways.

Keywords: A375.S2 melanoma cells; PEITC; ROS; apoptosis; mitochondria.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anticarcinogenic Agents / pharmacology*
  • Apoptosis / drug effects*
  • Blotting, Western
  • Calcium / metabolism
  • Cardiolipins / metabolism
  • Caspase 3 / metabolism
  • Cell Cycle / drug effects
  • Cell Line, Tumor
  • Cell Shape / drug effects
  • Comet Assay
  • DNA Damage
  • Flow Cytometry
  • Fluorescent Antibody Technique
  • Humans
  • Indicators and Reagents
  • Isothiocyanates / pharmacology*
  • Melanoma / drug therapy*
  • Melanoma / pathology
  • Membrane Potential, Mitochondrial / drug effects
  • Microscopy, Confocal
  • Mitochondria / drug effects
  • Mitochondria / metabolism*
  • Nitric Oxide / metabolism
  • Reactive Oxygen Species / metabolism*


  • Anticarcinogenic Agents
  • Cardiolipins
  • Indicators and Reagents
  • Isothiocyanates
  • Reactive Oxygen Species
  • Nitric Oxide
  • phenethyl isothiocyanate
  • Caspase 3
  • Calcium