Title: trans-3,4-dihydroxy-anti-1,2-epoxy-1,2,3,4-tetrahydrodi- benz[a,j]acridine involvement in dibenz[a,j]acridine DNA adduct formation in mouse skin consistent with Ha-ras mutation patterns in tumors.
Authors: Xue, W; Schneider, J; Mitchell, K; Jaeger, M; Nanayakkara, V; Talaska, G; Warshawsky, D
Published In Chem Res Toxicol, (2001 Jul)
Abstract: Dibenz[a,j]acridine (DBA), is a N-heteropolycyclic aromatic environmental carcinogen found in complex combustion mixtures. The major route of DBA metabolic activation is reportedly through the trans-3,4-dihydroxy-3,4-dihydroDBA (DBA-3,4-DHD). The present studies were undertaken to determine the role of trans-3,4-dihydroxy-anti-1,2-epoxy-1,2,3,4-tetrahydroDBA (DBADE) in DBA activation pathway(s), the DNA bases involved in the binding of DBA to DNA, and whether the adducts produced are consistent with the mutation pattern in the Ha-ras gene. DBA (300 microg) or 50 microg synthesized (+/-)-DBADE was applied to the back of female Hsd:ICR(Br) mice. The mice were sacrificed 48 h later, and skin DNA was isolated, hydrolyzed, and analyzed with (32)P-postlabeling. Of the four adducts produced in vivo, adduct 1 was the major adduct for DBA (>50%) and adduct 2 was the major adduct for DBADE (89%). After the reaction of (+/-)-DBADE with purine nucleotides or calf thymus (CT) DNA in vitro, 100% of the DBADE-2'-dAMP adducts and 94% of DBADE-CT DNA adducts were chromatographically identical on TLC with adduct 2 and 86% of the DBADE-2'-dGMP adducts were chromatographically consistent with adduct 1 by (32)P-postlabeling. Papillomas were induced on the backs of mice by a single application of 0.2 micromol of DBA followed by twice-weekly application of 12-o-tetra-decanoylphorbol-13-acetate (TPA, 2 microg) for 24-26 weeks. Skin carcinomas were induced by twice weekly applications of DBA (0.1 micromol) on the backs of mice. A to T and G to T transversions were found in codons 12, 13, and 61 of the Ha-ras gene in the treated mouse skin carcinoma and papilloma DNA. The mutational spectra in the Ha-ras gene are consistent with the DNA binding of DBA to dG or dA in vivo. Thus, this research has indicated that DBADE plays an important role in DBA metabolic activation and DNA binding in mouse skin, and an alternative pathway through a bis-dihydrodiol-epoxide of DBA may also be involved.
PubMed ID: 11453734
MeSH Terms: Acridines/metabolism; Acridines/toxicity*; Animals; Autoradiography; Benz(a)Anthracenes/metabolism*; Carcinogens/metabolism; Carcinogens/toxicity*; DNA Adducts/metabolism*; DNA, Neoplasm/genetics; DNA, Neoplasm/metabolism; Environmental Pollutants/toxicity; Female; Genes, ras*; Isotope Labeling; Mice; Mice, Inbred ICR; Models, Molecular; Mutation; Nucleotides/analysis; Papilloma/chemically induced; Papilloma/genetics*; Skin Neoplasms/chemically induced; Skin Neoplasms/genetics*