Oxidative stress in humans: validation of biomarkers of DNA damage.
Two studies have been performed to clarify the relationship between different markers of oxidative DNA damage commonly employed in molecular epidemiological studies. In the first, 8-Oxo-7,8-dihydroguanine (8-oxoGua) was induced in DNA of HeLa cells by treatment with different concentrations of photosensitizer Ro 19-8022 together with visible light. 8-OxoGua was estimated by the comet assay (alkaline single cell gel electrophoresis) with formamidopyrimidine DNA glycosylase and by HPLC with electrochemical detection. The dose-response curves indicate that the comet assay and HPLC are equally efficient at detecting induced damage. Background levels of 8-oxoGua in HeLa cells were 0.92 +/- 0.22 per 10(6) guanines by the comet assay and 2.09 +/- 0.13 per 10(6) guanines by HPLC. The second study was a small human trial, in which lymphocytes were collected for analysis of background levels of 8-oxoGua, as well as overnight and 24 h urine samples for measurement of excreted 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo) by ELISA. The mean level of 8-oxoGua in lymphocytes was determined as 1.33 +/- 0.21 per 10(6) guanines by the comet assay and 3.72 +/- 1.06 per 10(6) guanines by HPLC. A strong correlation was seen between overnight and 24 h urinary 8-oxodGuo (r = 0.93, P < 0.01). Overnight urinary 8-oxodGuo concentrations correlated with 8-oxoGua in lymphocytes measured by HPLC (r = 0.85, P < 0.05) or by the comet assay (r = 0.86, P < 0.05), although individual values from HPLC and the comet assay did not correlate with each other. It is reasonable to assess oxidative stress by any of these methods.
Rowett Research Institute, Greenburn Road, Bucksburn, Aberdeen AB21 9SB, UK.
Catherine M
CM
Gedik
Susanne P
SP
Boyle
Sharon G
SG
Wood
Nicholas J
NJ
Vaughan
Andrew R
AR
Collins
D000080242:8-Hydroxy-2'-Deoxyguanosine; D015415:Biomarkers / Q000032:analysis / Q000652:urine; D002851:Chromatography, High Pressure Liquid; D020552:Comet Assay; D004249:DNA Damage; D004273:DNA, Neoplasm / Q000378:metabolism*; D043382:DNA-Formamidopyrimidine Glycosylase; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000032:analysis* / Q000652:urine; D006367:HeLa Cells; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism; D009699:N-Glycosyl Hydrolases / Q000378:metabolism; D018384:Oxidative Stress / Q000502:physiology*; D015203:Reproducibility of Results
D015415:Biomarkers; D004273:DNA, Neoplasm; D000080242:8-Hydroxy-2'-Deoxyguanosine; D009699:N-Glycosyl Hydrolases; D043382:DNA-Formamidopyrimidine Glycosylase; D003849:Deoxyguanosine