Oligonucleotides in human urine do not contain 8-oxo-7,8-dihydrodeoxyguanosine.
The promutagenic DNA modification 8-oxo-7,8-dihydrodeoxyguanosine is the most frequently used marker for oxidative stress to DNA. The unmodified base and nucleoside and the 8-hydroxylated guanine base and nucleoside are found in urine, the latter used as a global measure of oxidative stress to DNA. Nucleotide excision repair (NER) excises a 27- to 29-mer oligonucleotide with oxidative lesions, and if found in urine, it could be used as a measure of DNA repair in vivo. Enzymatic hydrolysis of human urines followed by HPLC-tandem mass spectrometry was not able to reveal oligonucleotides and/or mononucleotides with the 8-oxo-7,8-dihydrodeoxyguanosine modification. The recovery of a synthetic oligonucleotide with the modification was complete (95% confidence limits: 98-124%). These experiments show that oligonucleotides are excreted into urine, but that 8-oxo-7,8-dihydrodeoxyguanosine is found only as the mononucleoside and is not present in any significant amounts in oligonucleotides. We conclude that oligonucleotides are excreted into urine, and they do not contain oxidized lesions. Either NER products are degraded after excision or NER functions differently in vivo in humans compared with cellular systems.
Weimann
Allan
A
Department of Clinical Pharmacology Q7642, Rigshospitalet, University Hospital Copenhagen, Blegdamsvej 9, DK-2100 Copenhagen, Denmark.
Riis
Bente
B
Poulsen
Henrik E
HE
D000080242:8-Hydroxy-2'-Deoxyguanosine; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000032:analysis*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D009841:Oligonucleotides / Q000737:chemistry / Q000652:urine*; D018384:Oxidative Stress
D009841:Oligonucleotides; D000080242:8-Hydroxy-2'-Deoxyguanosine; D003849:Deoxyguanosine