Free radicals-mediated induction of oxidized DNA bases and DNA-protein cross-links by nickel chloride.
Using the comet assay, we showed that nickel chloride at 250-1000 microM induced DNA damage in human lymphocytes, measured as the change in comet tail moment, which increased with nickel concentration up to 500 microM and then decreased. Observed increase might follow from the induction of strand breaks or/and alkali-labile sites (ALS) by nickel, whereas decrease from its induction of DNA-DNA and/or DNA-protein cross-links. Proteinase K caused an increase in the tail moment, suggesting that nickel chloride at 1000 microM might cross-link DNA with nuclear proteins. Lymphocytes exposed to NiCl(2) and treated with enzymes recognizing oxidized and alkylated bases: endonuclease III (Endo III), formamidopyrimidine-DNA glycosylase (Fpg) and 3-methyladenine-DNA glycosylase II (AlkA), displayed greater extent of DNA damage than those not treated with these enzymes, indicating the induction of oxidized and alkylated bases by nickel. The incubation of lymphocytes with spin traps, 5,5-dimethyl-pyrroline N-oxide (DMPO) and PBN decreased the extent of DNA damage, which might follow from the production of free radicals by nickel. The pre-treatment with Vitamin C at 10 microM and Vitamin E at 25 microM decreased the tail moment of the cells exposed to NiCl(2) at the concentrations of the metal causing strand breaks or/and ALS. The results obtained suggest that free radicals may be involved in the formation of strand breaks or/and ALS in DNA as well as DNA-protein cross-links induced by NiCl(2). Nickel chloride can also alkylate DNA bases. Our results support thesis on multiple, free radicals-based genotoxicity pathways of nickel.
Woźniak
Katarzyna
K
Department of Molecular Genetics, University of Lodz, Banacha 12/16, 90-237, Lodz, Poland. wozniak@biol.uni.lodz.pl
Błasiak
Janusz
J
D000328:Adult; D000975:Antioxidants / Q000494:pharmacology; D001205:Ascorbic Acid / Q000494:pharmacology; D002470:Cell Survival; D020552:Comet Assay; D003432:Cross-Linking Reagents; D004247:DNA / Q000187:drug effects / Q000378:metabolism*; D004249:DNA Damage; D004260:DNA Repair; D016166:Free Radical Scavengers / Q000494:pharmacology; D005609:Free Radicals / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008214:Lymphocytes / Q000187:drug effects* / Q000502:physiology; D008297:Male; D009532:Nickel / Q000494:pharmacology*; D016877:Oxidants / Q000494:pharmacology; D010084:Oxidation-Reduction; D014810:Vitamin E / Q000494:pharmacology
D000975:Antioxidants; D003432:Cross-Linking Reagents; D016166:Free Radical Scavengers; D005609:Free Radicals; D016877:Oxidants; D014810:Vitamin E; C022838:nickel chloride; D009532:Nickel; D004247:DNA; D006861:Hydrogen Peroxide; D001205:Ascorbic Acid