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.
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.
Lagadu
Stéphanie
S
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.
Pottier
Ivannah
I
Sichel
François
F
Laurent
Carine
C
Lefaix
Jean-Louis
JL
Prevost
Virginie
V
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
D015415:Biomarkers; D003470:Culture Media; D000080242:8-Hydroxy-2'-Deoxyguanosine; D003849:Deoxyguanosine