title: Radical production and DNA damage induced by carcinogenic 4-hydrazinobenzoic acid, an ingredient of mushroom Agaricus bisporus.
abstract: 4-Hydrazinobenzoic acid, an ingredient of mushroom Agaricus bisporus, is carcinogenic to rodents. To clarify the mechanism of carcinogenesis, we investigated DNA damage by 4-hydrazinobenzoic acid using (32)P-labeled DNA fragments obtained from the human p53 and p16 tumor suppressor genes. 4-Hydrazinobenzoic acid induced Cu(II)-dependent DNA damage especially piperidine-labile formation at thymine and cytosine residues. Typical hydroxyl radical scavengers showed no inhibitory effects on Cu(II)-mediated DNA damage by 4-hydrazinobenzoic acid. Bathocuproine and catalase inhibited the DNA damage, indicating the participation of Cu(I) and H(2)O(2) in the DNA damage. These findings suggest that H(2)O(2) generated by the autoxidation of 4-hydrazinobenzoic acid reacts with Cu(I) to form reactive oxygen species, capable of causing DNA damage. Interestingly, catalase did not completely inhibit DNA damage caused by a high concentration of 4-hydrazinobenzoic acid (over 50 microM) in the presence of Cu(II). 4-Hydrazinobenzoic acid induced piperidine-labile sites frequently at adenine and guanine residues in the presence of catalase. 4-Hydrazinobenzoic acid increased formation of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG), a characteristic oxidative DNA lesion, in calf thymus DNA, whereas 4-hydrazinobenzoic acid did not increase the formation of 8-oxodG in the presence of catalase. ESR spin-trapping experiments showed that the phenyl radical was formed during the reaction of 4-hydrazinobenzoic acid in the presence of Cu(II) and catalase. Matrix-assisted laser desorption/ionization time-of-flight mass (MALDI-TOF/mass) spectrometry analysis showed that phenyl radical formed adduct with adenosine and guanosine. These results suggested that 4-hydrazinobenzoic acid induced DNA damage via not only H(2)O(2) production but also phenyl radical production. This study suggests that both oxidative DNA damage and DNA adduct formation play important roles in the expression of carcinogenesis of 4-hydrazinobenzoic acid.
authors.affiliation: Department of Environmental and Molecular Medicine, Mie University Graduate School of Medicine, Japan.
authors.forename: Shinji
authors.initials: S
authors.lastname: Oikawa
authors.forename: Takahiro
authors.initials: T
authors.lastname: Ito
authors.forename: Michiko
authors.initials: M
authors.lastname: Iwayama
authors.forename: Shosuke
authors.initials: S
authors.lastname: Kawanishi
mesh_terms: D000080242:8-Hydroxy-2'-Deoxyguanosine; D000241:Adenosine / Q000378:metabolism; D000364:Agaricus / Q000737:chemistry*; D000818:Animals; D001565:Benzoates / Q000737:chemistry* / Q000633:toxicity*; D002273:Carcinogens / Q000737:chemistry* / Q000633:toxicity*; D002374:Catalase / Q000378:metabolism / Q000494:pharmacology; D002417:Cattle; D003300:Copper / Q000494:pharmacology; D004247:DNA / Q000187:drug effects / Q000378:metabolism; D018736:DNA Adducts / Q000096:biosynthesis; D004249:DNA Damage; D003849:Deoxyguanosine / Q000031:analogs & derivatives / Q000096:biosynthesis / Q000235:genetics; D004578:Electron Spin Resonance Spectroscopy / Q000379:methods; D016147:Genes, Tumor Suppressor / Q000187:drug effects; D006151:Guanosine / Q000378:metabolism; D006801:Humans; D017382:Reactive Oxygen Species / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013379:Substrate Specificity
chemical_list: D001565:Benzoates; D002273:Carcinogens; D018736:DNA Adducts; D017382:Reactive Oxygen Species; D006151:Guanosine; D003300:Copper; D000080242:8-Hydroxy-2'-Deoxyguanosine; D004247:DNA; C427203:calf thymus DNA; C053198:4-hydrazinobenzoic acid; D002374:Catalase; D003849:Deoxyguanosine; D000241:Adenosine