Initial levels of azoxymethane-induced DNA methyl adducts are not predictive of tumor susceptibility in inbred mice.
Inbred mice vary in susceptibility to colon carcinogens such as 1,2-dimethylhydrazine (DMH). Differential susceptibility may depend, in part, on formation of promutagenic DNA methyl adducts within target colonic mucosa. The present study was undertaken to evaluate the extent of DNA adduct formation in susceptible (SWR) and resistant (AKR) mice acutely exposed to the colon carcinogen azoxymethane (AOM), a direct metabolite of DMH. In the first experiment, 8-week-old SWR and AKR mice were treated i.p. with 20 mg/kg AOM and sacrificed 6 h later. DNA was isolated from distal colon and liver, and O6-methylguanine (O6-MeGua) adduct levels were assessed by immunoslot blot (ISB) analysis, using a monospecific antibody raised against O6-methyldeoxyguanosine. HPLC-fluorescence detection was also used to quantitate 06-MeGua and 7-methylguanine (7-MeGua), and to generate standard curves. At 6 h, both O6-MeGua and 7-MeGua were significantly higher (2- to 3-fold, p < 0.05) in AKR colon, while an opposite pattern was found in liver. In Experiment 2, mice were injected with AOM (20 mg/kg) and euthanized 12 and 48 h later. At 12 h, O6-MeGua levels were higher in colons (1.4-fold) of SWR mice. Forty-eight hours after treatment, however, adduct levels in colon were markedly (5-fold) reduced in SWR but were unchanged from 12 h in AKR. To further compare activation of AOM in both strains, colon microsomes were incubated with AOM and calf thymus DNA. Comparable levels of O6-MeGua were detected by ISB, demonstrating equivalent metabolic capacity in both SWR and AKR mice. These studies suggest that differential susceptibility to AOM-induced colon carcinogenesis is not based on initial target tissue DNA alkylation and unlikely to depend on differential metabolic capacity.
Papanikolaou
A
A
Department of Pharmaceutical Sciences, University of Connecticut, Storrs 06269-2092, USA.
Shank
R C
RC
Delker
D A
DA
Povey
A
A
Cooper
D P
DP
Rosenberg
D W
DW
D000818:Animals; D001397:Azoxymethane / Q000633:toxicity*; D001711:Biotransformation; D002273:Carcinogens / Q000494:pharmacology / Q000633:toxicity*; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000032:analysis / Q000187:drug effects / Q000302:isolation & purification; D018736:DNA Adducts / Q000187:drug effects / Q000378:metabolism*; D006147:Guanine / Q000031:analogs & derivatives / Q000378:metabolism; D015151:Immunoblotting; D008297:Male; D008770:Methylnitrosourea / Q000494:pharmacology; D051379:Mice; D008806:Mice, Inbred AKR; D008815:Mice, Inbred Strains; D009374:Neoplasms, Experimental / Q000139:chemically induced*; D011237:Predictive Value of Tests
D002273:Carcinogens; D018736:DNA Adducts; D006147:Guanine; C008450:7-methylguanine; D008770:Methylnitrosourea; D004247:DNA; C008449:O-(6)-methylguanine; D001397:Azoxymethane