title: Detection of extracellular 8-oxo-7,8-dihydro-2'-deoxyguanosine as a biomarker of oxidative damage in X-irradiated fibroblast cultures: optimization of analytical procedure.
abstract: We have developed a simple methodology, based on single-step solid-phase extraction followed by isocratic high-performance liquid chromatography coupled with electrochemical detection (HPLC-ECD), to determine extracellular 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) in culture supernatants of normal human dermal fibroblasts. A standard addition method, using externally added 8-oxodG (0.5 and 1 pmol) was employed to eliminate matrix effects arising from the chemically complex, protein-rich medium. Secondly, applying this procedure to X-ray irradiated fibroblasts, we report a significant twofold increase in the levels of 8-oxodG at the radiobiologically relevant dose of 6 Gy. This suggests that extracellular 8-oxodG might be a useful biomarker for oxidative stress following moderate doses of X-irradiation.
authors.affiliation: Groupe Régional d'Etudes sur le Cancer - UPRES EA 1772 - IFR 146, Université de Caen Basse-Normandie and Centre de Lutte Contre le Cancer François Baclesse, Caen, France.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Lagadu
authors.forename: Ivannah
authors.initials: I
authors.lastname: Pottier
authors.forename: François
authors.initials: F
authors.lastname: Sichel
authors.forename: Carine
authors.initials: C
authors.lastname: Laurent
authors.forename: Jean-Louis
authors.initials: JL
authors.lastname: Lefaix
authors.forename: Virginie
authors.initials: V
authors.lastname: Prevost
mesh_terms: D000080242:8-Hydroxy-2'-Deoxyguanosine; D015415:Biomarkers / Q000378:metabolism*; D002138:Calibration; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D003470:Culture Media; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000378:metabolism; D004563:Electrochemistry; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism / Q000528:radiation effects*; D006801:Humans; D018384:Oxidative Stress; D014965:X-Rays
chemical_list: D015415:Biomarkers; D003470:Culture Media; D000080242:8-Hydroxy-2'-Deoxyguanosine; D003849:Deoxyguanosine