Effect of lipophilic chelators on oxyradical-induced DNA strand breaks in human granulocytes: paradoxical effect of 1,10-phenanthroline.
Strand breaks can be produced in the DNA of intact granulocytes by a flux of oxyradicals (O2- and H2O2) generated by tetradecanoylphorbol acetate (TPA) or by a flux of H2O2 generated by glucose oxidase. The mechanism by which such breaks are induced is still uncertain. Lipophilic chelators such as dipyridyl and 1,10-phenanthroline (OP) strongly inhibit strand breaks induced by H2O2, presumably because of their ability to chelate intracellular iron. We now report that dipyridyl also partially inhibits strand breaks in TPA-stimulated granulocytes while a "copper-specific" lipophilic chelator, neocuproine, has no effect. As opposed to these effects, OP increases the number of strand breaks in TPA-stimulated granulocytes. Superoxide dismutase (SOD) (but not catalase) partially blocks this increase. Both the cell-impermeable chelator, EDTA, and neocuproine strongly block the increase also. In fact, in the presence of EDTA, OP behaves like dipyridyl and inhibits strand breaks. Preformed OP2-copper(II) complex causes DNA breaks in TPA-stimulated granulocytes. The paradoxical effect of OP may be explained by assuming that OP may form two different metal complexes, a DNA-damaging complex with copper or an inhibitory complex with iron. If copper(II) and O2- are present, the first complex may form and the net effect may be an increase in strand breaks. If the formation of this complex is prevented by SOD, EDTA, or neocuproine, then OP may complex iron and the net effect may be (like dipyridyl) an inhibition of strand breaks. The source of the copper responsible for the formation of OP2-copper complex is unknown.
Birnboim
H C
HC
Department of Experimental Oncology, Ottawa Regional Cancer Centre, Ontario, Canada.
D015082:2,2'-Dipyridyl / Q000494:pharmacology; D002614:Chelating Agents / Q000494:pharmacology*; D003300:Copper / Q000378:metabolism; D004247:DNA / Q000187:drug effects*; D004249:DNA Damage / Q000187:drug effects*; D004492:Edetic Acid / Q000494:pharmacology; D005609:Free Radicals; D006098:Granulocytes / Q000737:chemistry*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D010618:Phenanthrolines / Q000494:pharmacology*; D013482:Superoxide Dismutase / Q000494:pharmacology; D013481:Superoxides / Q000494:pharmacology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D002614:Chelating Agents; D005609:Free Radicals; D010618:Phenanthrolines; D013481:Superoxides; D015082:2,2'-Dipyridyl; D003300:Copper; C002701:neocuproine; D004247:DNA; D004492:Edetic Acid; D006861:Hydrogen Peroxide; D013482:Superoxide Dismutase; D013755:Tetradecanoylphorbol Acetate; C025205:1,10-phenanthroline