Rapid human melanoma cell death induced by sanguinarine through oxidative stress

Eur J Pharmacol. 2013 Apr 5;705(1-3):109-18. doi: 10.1016/j.ejphar.2013.02.035. Epub 2013 Mar 13.

Abstract

Sanguinarine is a natural isoquinoline alkaloid derived from the root of Sanguinaria canadensis and from other poppy fumaria species, and is known to have a broad spectrum of pharmacological properties. Here we have found that sanguinarine, at low micromolar concentrations, showed a remarkably rapid killing activity against human melanoma cells. Time-lapse videomicroscopy showed rapid morphological changes compatible with an apoptotic cell death, which was further supported by biochemical markers, including caspase activation, poly(ADP-ribose) polymerase (PARP) cleavage and DNA breakdown. Pan-caspase inhibition blocked sanguinarine-induced cell death. Sanguinarine treatment also induced an increase in intracellular calcium concentration, which was inhibited by dantrolene, and promoted cleavage of BAP-31, thus suggesting a putative role for Ca(2+) release from endoplasmic reticulum and a cross-talk between endoplasmic reticulum and mitochondria in the anti-melanoma action of sanguinarine. Sanguinarine disrupted the mitochondrial transmembrane potential (ΔΨm), released cytochrome c and Smac/DIABLO from mitochondria to cytosol, and induced oxidative stress. Overexpression of Bcl-XL by gene transfer did not prevent sanguinarine-mediated cell death, oxidative stress or release of mitochondrial apoptogenic proteins. However, preincubation with N-acetyl-l-cysteine (NAC) prevented sanguinarine-induced oxidative stress, PARP cleavage, release of apoptogenic mitochondrial proteins, and cell death. Pretreatment with glutathione (GSH) also inhibited the anti-melanoma activity of sanguinarine. Thus, pretreatment with the thiol antioxidants NAC and GSH abrogated the killing activity of sanguinarine. Taking together, our data indicate that sanguinarine is a very rapid inducer of human melanoma caspase-dependent cell death that is mediated by oxidative stress.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylcysteine / pharmacology
  • Antineoplastic Agents / pharmacology*
  • Benzophenanthridines / pharmacology*
  • Calcium / metabolism
  • Caspase 3 / metabolism
  • Caspase 7 / metabolism
  • Cell Death / drug effects
  • Cell Line, Tumor
  • Cell Survival / drug effects
  • Glutathione / pharmacology
  • Humans
  • Isoquinolines / pharmacology*
  • Melanoma / drug therapy*
  • Melanoma / metabolism
  • Melanoma / physiopathology
  • Membrane Potential, Mitochondrial / drug effects
  • Membrane Proteins / metabolism
  • Mitochondria / drug effects
  • Mitochondria / physiology
  • Oxidative Stress / drug effects
  • Reactive Oxygen Species / metabolism
  • bcl-X Protein / metabolism

Substances

  • Antineoplastic Agents
  • BCAP31 protein, human
  • BCL2L1 protein, human
  • Benzophenanthridines
  • Isoquinolines
  • Membrane Proteins
  • Reactive Oxygen Species
  • bcl-X Protein
  • sanguinarine
  • Caspase 3
  • Caspase 7
  • Glutathione
  • Calcium
  • Acetylcysteine