Rapid human melanoma cell death induced by sanguinarine through oxidative stress.

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.

• Authors: Ana Burgeiro, Ana C Bento, Consuelo Gajate, Paulo J Oliveira, Faustino Mollinedo
• Journal: European journal of pharmacology (Eur J Pharmacol) Vol. 705 Issue 1-3 Pg. 109-18 (Apr 05 2013) ISSN: 1879-0712 [Electronic] Netherlands
• PMID: 23499690 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2013 Elsevier B.V. All rights reserved.

Chemical References

• 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

Topics

• 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, metabolism, physiopathology)
• Membrane Potential, Mitochondrial (drug effects)
• Membrane Proteins (metabolism)
• Mitochondria (drug effects, physiology)
• Oxidative Stress (drug effects)
• Reactive Oxygen Species (metabolism)
• bcl-X Protein (metabolism)