Abstract
Therapeutic induction of apoptosis is an important goal of anticancer drug design. Cellular carbonyl stress mediated by endogenous reactive carbonyl species (RCS) such as glyoxal and methylglyoxal (MG) affects proliferative signaling and metastasis of human tumor cells. Recent research suggests that RCS produced constitutively during increased tumor cell glycolysis may be antiapoptotic survival factors and thus represent a novel molecular target for anticancer intervention. Here, we demonstrate the tumor cell-specific apoptogenicity of carbonyl scavengers, which act by covalently trapping RCS, against human (A375, G361, and LOX) and murine (B16) melanoma cell lines. A structure-activity relationship study identified nucleophilic carbonyl scavenger pharmacophores as the functional determinants of apoptogenic antimelanoma activity of structurally diverse agents such as 3,3-dimethyl-d-cysteine and aminoguanidine. Previous work has demonstrated that covalent adduction of protein-arginine residues in the mitochondrial permeability transition (MPT) pore and heat shock protein 27 by intracellular MG produced in tumor cell glycolysis inhibits mitochondrial apoptosis and enhances cancer cell survival. Indeed, in various melanoma cell lines, carbonyl scavenger-induced apoptosis was antagonized by pretreatment with the membrane-permeable RCS phenylglyoxal (PG). Carbonyl scavenger-induced apoptosis was associated with early loss of mitochondrial transmembrane potential, and cyclosporin A antagonized the effects of carbonyl scavengers, suggesting a causative role of MPT pore opening in carbonyl scavenger apoptogenicity. Consistent with RCS inhibition of mitochondrial apoptosis in melanoma cells, staurosporine-induced apoptosis also was suppressed by PG pretreatment. Our results suggest that carbonyl scavengers acting as direct molecular antagonists of RCS are promising apoptogenic prototype agents for antimelanoma drug design.
Footnotes
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This study was supported in part by National Institutes of Health Grants CA43894, CA99469, CA106677, PO1CA27502, P30-ESO6694, and P30CA023074 and by Niadyne, Inc. (Tucson, AZ).
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doi:10.1124/jpet.105.094953.
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ABBREVIATIONS: RCS, reactive carbonyl species; MG, methylglyoxal; AGE, advanced glycation end products; MPT, mitochondrial permeability transition; Hsp, heat shock protein; MEC, 3-methyl-3-ethyl-d-cysteine; BCS, bovine calf serum; FITC, fluorescein isothiocyanate; PI, propidium iodide; Δψm, mitochondrial transmembrane potential; JC-1, 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolyl-carbocyanine iodide; PBS, phosphate-buffered saline; DMC, 3,3-dimethyl-d-cysteine; SAR, structure-activity relationship; ADMC, N-acetyl-3,3-dimethyl-d-cysteine; NAC, Nα-acetyl-l-cysteine; DMCSS, 3,3-dimethyl-d-cysteine-disulfide; AG, aminoguanidine; SC, semicarbazide hydrochloride; DMBG, 1,1-dimethylbiguanide hydrochloride; PG, phenylglyoxal; AV, Annexin V; DMCM, 3,3-dimethyl-d-cysteine-methylester.
- Received August 30, 2005.
- Accepted October 5, 2005.
- The American Society for Pharmacology and Experimental Therapeutics
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