title: DNA damage induced by methylated trivalent arsenicals is mediated by reactive oxygen species.
abstract: Arsenic is a human carcinogen; however, the mechanisms of arsenic's induction of carcinogenic effects have not been identified clearly. We have shown previously that monomethylarsonous acid (MMA(III)) and dimethylarsinous acid (DMA(III)) are genotoxic and can damage supercoiled phiX174 DNA and the DNA in peripheral human lymphocytes in culture. These trivalent arsenicals are biomethylated forms of inorganic arsenic and have been detected in the urine of subjects exposed to arsenite and arsenate. We show here by molecular, chemical, and physical methods that reactive oxygen species (ROS) are intermediates in the DNA-damaging activities of MMA(III) and DMA(III). Using the phiX174 DNA nicking assay we found that the ROS inhibitors Tiron, melatonin, and the vitamin E analogue Trolox inhibited the DNA-nicking activities of both MMA(III) and DMA(III) at low micromolar concentrations. The spin trap agent 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) also was effective at preventing the DNA nicking induced by MMA(III) and DMA(III). ESR spectroscopy studies using DMPO identified a radical as a ROS intermediate in the DNA incubations with DMA(III). This radical adduct was assigned to the DMPO-hydroxyl free radical adduct on the basis of comparison of the observed hyperfine splitting constants and line widths with those reported in the literature. The formation of the DMPO-hydroxyl free radical adduct was dependent on time and the presence of DMA(III) and was completely inhibited by Tiron and Trolox and partially inhibited by DMSO. Using electrospray mass spectrometry, micromolar concentrations of DMA(V) were detected in the DNA incubation mixtures with DMA(III). These data are consistent with the conclusions that the DNA-damaging activity of DMA(III) is an indirect genotoxic effect mediated by ROS-formed concomitantly with the oxidation of DMA(III) to DMA(V).
authors.affiliation: Environmental Carcinogenesis Division, National Health and Environmental Effects Research Laboratory, Office of Research and Development, U.S. Environmental Protection Agency, Research Triangle Park, North Carolina 27711, USA. nesnow.stephen@epa.gov
authors.forename: Stephen
authors.initials: S
authors.lastname: Nesnow
authors.forename: Barbara C
authors.initials: BC
authors.lastname: Roop
authors.forename: Guy
authors.initials: G
authors.lastname: Lambert
authors.forename: Maria
authors.initials: M
authors.lastname: Kadiiska
authors.forename: Ronald P
authors.initials: RP
authors.lastname: Mason
authors.forename: William R
authors.initials: WR
authors.lastname: Cullen
authors.forename: Marc J
authors.initials: MJ
authors.lastname: Mass
mesh_terms: D001151:Arsenic / Q000633:toxicity*; D010584:Bacteriophage phi X 174 / Q000737:chemistry / Q000187:drug effects; D002101:Cacodylic Acid / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*; D003497:Cyclic N-Oxides / Q000737:chemistry / Q000494:pharmacology; D004249:DNA Damage; D019175:DNA Methylation / Q000187:drug effects*; D004278:DNA, Superhelical / Q000187:drug effects; D004578:Electron Spin Resonance Spectroscopy; D016166:Free Radical Scavengers / Q000494:pharmacology; D006801:Humans; D020287:In Situ Nick-End Labeling / Q000379:methods; D009942:Organometallic Compounds / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*; D017382:Reactive Oxygen Species / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism* / Q000633:toxicity*; D021241:Spectrometry, Mass, Electrospray Ionization; D013997:Time Factors
chemical_list: D003497:Cyclic N-Oxides; D004278:DNA, Superhelical; D016166:Free Radical Scavengers; D009942:Organometallic Compounds; D017382:Reactive Oxygen Species; C472511:dimethylarsinous acid; C406082:monomethylarsonous acid; C017245:5,5-dimethyl-1-pyrroline-1-oxide; D002101:Cacodylic Acid; D001151:Arsenic