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Titolo:
IN-VITRO INHIBITION BY N-ACETYLCYSTEINE OF OXIDATIVE DNA MODIFICATIONS DETECTED BY P-32 POSTLABELING
Autore:
IZZOTTI A; ORLANDO M; GASPARINI L; SCATOLINI L; CARTIGLIA C; TULIMIERO L; DEFLORA S;
Indirizzi:
UNIV GENOA,INST HYG & PREVENT MED,VIA PASTORE 1 I-16132 GENOA ITALY UNIV GENOA,INST HYG & PREVENT MED I-16132 GENOA ITALY
Titolo Testata:
Free radical research
fascicolo: 2, volume: 28, anno: 1998,
pagine: 165 - 178
SICI:
1071-5762(1998)28:2<165:IIBNOO>2.0.ZU;2-X
Fonte:
ISI
Lingua:
ENG
Soggetto:
HYDROGEN-PEROXIDE; I-COMPOUNDS; OXYGEN RADICALS; MAMMALIAN-CELLS; DAMAGE INVITRO; ADDUCTS; MUTAGENICITY; GENOTOXICITY; SENSITIVITY; ANTIOXIDANT;
Keywords:
REACTIVE OXYGEN SPECIES; HYDROGEN PEROXIDE; HYDROXYL RADICAL; N-ACETYLCYSTEINE; DNA DAMAGE; P-32 POST-LABELING;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
36
Recensione:
Indirizzi per estratti:
Citazione:
A. Izzotti et al., "IN-VITRO INHIBITION BY N-ACETYLCYSTEINE OF OXIDATIVE DNA MODIFICATIONS DETECTED BY P-32 POSTLABELING", Free radical research, 28(2), 1998, pp. 165-178

Abstract

Reactive oxygen species are involved in the pathogenesis of cancer and other chronic degenerative diseases through a variety of mechanisms,including DNA damage. We investigated by P-32 and P-33 postlabeling analyses the nucleotidic modifications induced in vitro by treating calf thymus DNA with H2O2 and CuSO4, interacting in a Fenton type reaction. Six different enrichment procedures and three chromatographic systems were comparatively assayed. The chromatographic system using phosphate/urea, which is more suitable for detecting bulky DNA adducts, was rather insensitive. In contrast, the system using acetic acid/ammoniumformate revealed high levels of mononucleotidic modifications. In terms of ratio of adduct levels in treated and untreated DNA, the enrichment procedures ranked as follows: nuclease P-1 (19.6), no enrichment (18.3), digestion to trinucleotides (17.6), digestion to monophosphate mononucleotides (8.4), digestion to dinucleotides (3.4), and extraction with butanol (<1.0). The system using formic acid/ammonium formate was quite efficient in detecting 8-hydroxy-2'-deoxyguanosine. Labeling with P-33 fur ther enhanced the sensitivity of the method. The oxidative damage was so intense to produce a strong DNA fragmentation detectable by agarose gel electrophoresis, and nucleotidic modifications weremore intense when DNA fragmentation was greater. The DNA alterations produced by H2O2 alone were significantly lower than those produced following reaction of H2O2 with CuSO4. The thiol N-acetylcysteine (NAC) was quite efficient in inhibiting both nucleotidic modifications and DNA fragmentation produced in vitro by either H2O2 Or the OH generatingsystem. These results support at a molecular level the findings of previous studies showing the ability of NAC to inhibit the genotoxicity of peroxides and of reactive oxygen species generated by electron transfer reactions.

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Documento generato il 02/07/20 alle ore 22:39:49