Detection of 1,N6-ethenodeoxyadenosine and 3,N4-ethenodeoxycytidine by immunoaffinity/32P-postlabelling in liver and lung DNA of mice treated with ethyl carbamate (urethane) or its metabolites.
The capacity of the chemical carcinogen ethyl carbamate (EC, urethane) and its metabolites vinyl carbamate (VC) and vinyl carbamate epoxide (VCO) to form ethenobases was studied in liver and lung DNA of 12-day-old and adult CD-1, B6C3F1, C3H/HeJ and C57BL/6J mice. Following single and multiple doses of EC, VC or VCO, the formation of 1,N6-ethenodeoxyadenosine (epsilon dA) and 3,N4-ethenodeoxycytidine (epsilon dC) was quantified by an immunoaffinity chromatography/32P-postlabelling technique. Both etheno adducts were detected in untreated control DNA samples from liver and lung in the range of 2-15 adducts/10(9) parent nucleotides. Following five repeated injections of 250 or 280 nmol/g body wt VC to adult mice, 51, 57 and 78 epsilon dA/10(9) dA and 28, 42 and 42 epsilon dC/10(9) dC (means of duplicate analyses) were detected in liver DNA of CD-1, C3H/HeJ and C57BL/6J mice respectively. In lung DNA of these VC-treated mice, the levels were 87, 49 and 58 (epsilon dA/10(9) dA) and 64, 39 and 43 (epsilon dC/10(9) dC) respectively. Under similar dose regimens, lower levels of etheno adducts were detected in B6C3F1 mice. Etheno-DNA adducts were also formed in liver and lung upon treatment with EC in adult mice, but at 3-fold lower levels as compared with VC. In 12-day-old C3H/HeJ and C57BL/6J mice, 2- to 3-fold higher etheno adduct levels were detected in liver DNA, when compared with adults, upon a single treatment with 250 nmol/g body wt VC, suggesting that young animals are more susceptible to adduct formation. Combined analysis of adduct formation in adult CD-1, C3H/HeJ and C57BL/6J mice at the higher dose showed a statistically significant increase in etheno adduct formation in the order EC > VC. The results demonstrate that EC and its activated intermediates bind to liver and lung DNA to form epsilon dA and epsilon dC, and the differences in DNA binding further support the hypothesis that metabolic activation of EC to VC is involved. Preliminary data also suggest that background levels of epsilon dA and epsilon dC in DNA are affected by the type of diet given to the animals.
Fernando
R C
RC
German Cancer Research Center, Division of Toxicology and Cancer Risk Factors, Heidelberg, Germany.
Nair
J
J
Barbin
A
A
Miller
J A
JA
Bartsch
H
H
D000818:Animals; D002273:Carcinogens / Q000378:metabolism / Q000633:toxicity; D002846:Chromatography, Affinity / Q000379:methods; D004247:DNA / Q000737:chemistry*; D018736:DNA Adducts / Q000032:analysis; D003839:Deoxyadenosines / Q000032:analysis*; D003841:Deoxycytidine / Q000031:analogs & derivatives* / Q000032:analysis; D005260:Female; D008099:Liver / Q000737:chemistry*; D008168:Lung / Q000737:chemistry*; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D010761:Phosphorus Radioisotopes; D013045:Species Specificity; D013056:Spectrophotometry, Ultraviolet; D014520:Urethane / Q000031:analogs & derivatives / Q000378:metabolism / Q000633:toxicity*
D002273:Carcinogens; D018736:DNA Adducts; D003839:Deoxyadenosines; D010761:Phosphorus Radioisotopes; D003841:Deoxycytidine; D014520:Urethane; C032221:1,N(6)-ethenodeoxyadenosine; C032222:3,N(4)-ethenodeoxycytidine; D004247:DNA