Detection of oxidative damage by 32P-postlabelling: 8-hydroxydeoxyguanosine as a marker of exposure.

2.50
Hdl Handle:
http://hdl.handle.net/10541/100384
Title:
Detection of oxidative damage by 32P-postlabelling: 8-hydroxydeoxyguanosine as a marker of exposure.
Authors:
Povey, Andrew C; Wilson, V L; Weston, A; Doan, V T; Wood, M L; Essigmann, J M; Shields, P G
Abstract:
Human exposure to reactive oxygen species is unavoidable and has been implicated in the etiology of a number of human diseases. This exposure results in the formation of various modified DNA bases: the promutagenic lesion 8-hydroxydeoxyguanosine (8OHdG), in particular, is a major product. We have developed an assay using ion-pair HPLC and 32P-postlabelling to quantify 8OHdG in human DNA with high specificity and sensitivity. An internal standard is used to account for variations in labelling efficiency. Chemically synthesized 8OHdG 3'-monophosphate and 5'-monophosphate standards were used to optimize the HPLC-32P-postlabelling and TLC separative steps, respectively. The assay was validated using known ratios of 8OHdG to normal nucleotides. The limit of detection is in the range of one 8OHdG residue per 10(6)-10(7) dG residues. Using this procedure, 8OHdG levels of 16-35 8OHdG adducts per 10(5) dG residues have been found in leukocytes isolated from patients who received 180-200 cGy of ionizing radiation. These levels were 2-4-fold greater than those found in an unexposed individual. Since 8OHdG may be formed during DNA extraction and digestion, current procedures for measuring background levels are discussed.
Affiliation:
Cancer Research Campaign Department of Carcinogenesis, Paterson Institute for Cancer Research, Manchester, UK.
Citation:
Detection of oxidative damage by 32P-postlabelling: 8-hydroxydeoxyguanosine as a marker of exposure. 1993 (124):105-14 IARC Sci. Publ.
Journal:
IARC Scientific Publications
Issue Date:
1993
URI:
http://hdl.handle.net/10541/100384
PubMed ID:
8225472
Type:
Article
Language:
en
ISSN:
0300-5038
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorPovey, Andrew Cen
dc.contributor.authorWilson, V Len
dc.contributor.authorWeston, Aen
dc.contributor.authorDoan, V Ten
dc.contributor.authorWood, M Len
dc.contributor.authorEssigmann, J Men
dc.contributor.authorShields, P Gen
dc.date.accessioned2010-06-07T10:44:48Z-
dc.date.available2010-06-07T10:44:48Z-
dc.date.issued1993-
dc.identifier.citationDetection of oxidative damage by 32P-postlabelling: 8-hydroxydeoxyguanosine as a marker of exposure. 1993 (124):105-14 IARC Sci. Publ.en
dc.identifier.issn0300-5038-
dc.identifier.pmid8225472-
dc.identifier.urihttp://hdl.handle.net/10541/100384-
dc.description.abstractHuman exposure to reactive oxygen species is unavoidable and has been implicated in the etiology of a number of human diseases. This exposure results in the formation of various modified DNA bases: the promutagenic lesion 8-hydroxydeoxyguanosine (8OHdG), in particular, is a major product. We have developed an assay using ion-pair HPLC and 32P-postlabelling to quantify 8OHdG in human DNA with high specificity and sensitivity. An internal standard is used to account for variations in labelling efficiency. Chemically synthesized 8OHdG 3'-monophosphate and 5'-monophosphate standards were used to optimize the HPLC-32P-postlabelling and TLC separative steps, respectively. The assay was validated using known ratios of 8OHdG to normal nucleotides. The limit of detection is in the range of one 8OHdG residue per 10(6)-10(7) dG residues. Using this procedure, 8OHdG levels of 16-35 8OHdG adducts per 10(5) dG residues have been found in leukocytes isolated from patients who received 180-200 cGy of ionizing radiation. These levels were 2-4-fold greater than those found in an unexposed individual. Since 8OHdG may be formed during DNA extraction and digestion, current procedures for measuring background levels are discussed.en
dc.language.isoenen
dc.subject.meshBiological Markers-
dc.subject.meshCarcinogens, Environmental-
dc.subject.meshDNA-
dc.subject.meshDNA Damage-
dc.subject.meshDeoxyguanosine-
dc.subject.meshEnvironmental Exposure-
dc.subject.meshHumans-
dc.subject.meshOligonucleotides-
dc.subject.meshOxidation-Reduction-
dc.subject.meshPhosphorus Radioisotopes-
dc.subject.meshReactive Oxygen Species-
dc.titleDetection of oxidative damage by 32P-postlabelling: 8-hydroxydeoxyguanosine as a marker of exposure.en
dc.typeArticleen
dc.contributor.departmentCancer Research Campaign Department of Carcinogenesis, Paterson Institute for Cancer Research, Manchester, UK.en
dc.identifier.journalIARC Scientific Publicationsen

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