Cytotoxic activity of sanguinarine and dihydrosanguinarine in human promyelocytic leukemia HL-60 cells.

Abstract

The benzo[c]phenanthridine alkaloid sanguinarine has been studied for its antiproliferative activity in many cell types. Almost nothing however, is known about the cytotoxic effects of dihydrosanguinarine, a metabolite of sanguinarine. We compared the cytotoxicity of sanguinarine and dihydrosanguinarine in human leukemia HL-60 cells. Sanguinarine produced a dose-dependent decline in cell viability with IC(50) (inhibitor concentration required for 50% inhibition of cell viability) of 0.9 microM as determined by MTT assay after 4h exposure. Dihydrosanguinarine showed much less cytotoxicity than sanguinarine: at the highest concentration tested (20 microM) and 24h exposure, dihydrosanguinarine decreased viability only to 52%. Cytotoxic effects of both alkaloids were accompanied by activation of the intrinsic apoptotic pathway since we observed the dissipation of mitochondrial membrane potential, induction of caspase-9 and -3 activities, the appearance of sub-G(1) DNA and loss of plasma membrane asymmetry. This aside, sanguinarine also increased the activity of caspase-8. As shown by flow cytometry using annexin V/propidium iodide staining, 0.5 microM sanguinarine induced apoptosis while 1-4 microM sanguinarine caused necrotic cell death. In contrast, dihydrosanguinarine at concentrations from 5 microM induced primarily necrosis, whereas apoptosis occurred at 10 microM and above. We conclude that both alkaloids may cause, depending on the alkaloid concentration, both necrosis and apoptosis of HL-60 cells.

• Authors: Jirí Vrba, Petr Dolezel, Jaroslav Vicar, Jitka Ulrichová
• Journal: Toxicology in vitro : an international journal published in association with BIBRA (Toxicol In Vitro) Vol. 23 Issue 4 Pg. 580-8 (Jun 2009) ISSN: 0887-2333 [Print] England
• PMID: 19346183 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Benzophenanthridines
• Isoquinolines
• dihydrosanguinarine
• sanguinarine
• Caspases

Topics

• Apoptosis (drug effects)
• Benzophenanthridines (pharmacology)
• Caspases (metabolism)
• Cell Cycle (drug effects)
• Cell Survival (drug effects)
• HL-60 Cells
• Humans
• Isoquinolines (pharmacology)
• Membrane Potential, Mitochondrial (drug effects)