The formation of 2-aminofluorene-DNA adducts in vivo: evidence for peroxidase-mediated activation.
Formation of DNA adducts in various tissues of dogs fed a single dose of the carcinogen 2-aminofluorene was investigated. Adduct analysis was performed using a technique that allows measurement of both N-(deoxyguanosin-8-yl)-2-amino-2-aminofluorene-DNA adduct formed by reaction of N-hydroxy-2-aminofluorene with DNA, as well as the polar 2-aminofluorene-DNA adducts formed when 2-aminofluorene is activated by prostaglandin H synthase-peroxidase in vitro. Two male beagle (A and B) dogs were examined and a different DNA adduct profile was observed with each dog. For the dog A, N-(deoxyguanosin-8-yl)-2-aminofluorene was the major adduct found in hepatic DNA; no peroxidase-derived adducts were detected in this tissue. In contrast, adducts eluting similarly to peroxidase-derived adducts were found in urinary tract tissues of this dog with the relative abundance of these adducts in the order urothelium greater than renal medulla greater than renal cortex, which correlates with the respective tissues' prostaglandin H synthase activity. N-(Deoxyguanosin-8-yl)-2-aminofluorene was detected in the renal tissues, but not in urothelium. For dog B, only the N-(deoxyguanosin-8-yl)-2-aminofluorene adduct was observed in all tissues examined, including the urothelium. However, total binding to liver, kidney, and bladder were two-, two-, and four-fold lower, respectively, than dog A. These data indicate that both prostaglandin H synthase-mediated activation and N-hydroxylation of 2-aminofluorene occur in vivo and may be subjected to pharmacodynamic considerations. Furthermore, the tissue distribution of the peroxidase-mediated 2-aminofluorene adducts suggests this process may also be of importance in the bladder-specific carcinogenicity of aromatic amines.
Krauss
R S
RS
Laboratory of Molecular Biophysics, National Institute of Environmental Health Sciences, Research Triangle Park, North Carolina 27709.
Angerman-Stewart
J
J
Eling
T E
TE
Dooley
K L
KL
Kadlubar
F F
FF
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000032:analysis*; D018736:DNA Adducts; D004249:DNA Damage; D004285:Dogs; D004789:Enzyme Activation; D004848:Epithelium / Q000187:drug effects / Q000378:metabolism; D005449:Fluorenes / Q000032:analysis* / Q000378:metabolism / Q000633:toxicity*; D007668:Kidney / Q000187:drug effects / Q000378:metabolism; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology; D006899:Mixed Function Oxygenases / Q000378:metabolism; D009153:Mutagens / Q000378:metabolism / Q000633:toxicity*; D010544:Peroxidases / Q000378:metabolism*; D001743:Urinary Bladder / Q000187:drug effects / Q000378:metabolism
C052668:2-aminofluorene-DNA complex; D018736:DNA Adducts; D005449:Fluorenes; D009153:Mutagens; C012177:2-aminofluorene; D004247:DNA; D006899:Mixed Function Oxygenases; D010544:Peroxidases