PMID: 9645393Jun 30, 1998Paper

In vitro inhibition by N-acetylcysteine of oxidative DNA modifications detected by 32P postlabeling

Free Radical Research
Alberto IzzottiSilvio De Flora

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 32p and 33P 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/ammonium formate 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 P1 (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 33p further enhanced the sensitivity of the method. The oxidative damage was so intense to produce a strong DNA fragmentation ...Continue Reading

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Citations

Nov 10, 2000·International Journal of Cancer. Journal International Du Cancer·F D'AgostiniS de Flora
Apr 1, 1999·Environmental Toxicology and Pharmacology·E PinelliA Pfohl-Leszkowicz
Mar 24, 2005·European Journal of Cancer Prevention : the Official Journal of the European Cancer Prevention Organisation (ECP)·S-K ChangS-Y Hong
Jan 31, 2006·Journal of Pineal Research·Katarzyna WiniarskaJadwiga Bryla
Mar 17, 2015·Mutation Research·Alberto IzzottiSergio Claudio Saccà
Jun 12, 2003·The American Journal of Medicine·Alberto IzzottiSilvio De Flora
Dec 25, 2004·Carcinogenesis·Francesco D'AgostiniSilvio De Flora
Apr 24, 2003·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Alberto IzzottiSilvio De Flora
Mar 22, 2001·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·A IzzottiS De Flora

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