TY - JOUR
T1 - Covalent binding of (+) 7S-trans-7,8-dihydrobenzo [a]pyrene-7,8-diol to trout DNA
T2 - P-450- and peroxidation-dependent pathways
AU - Kelly, Jack D.
AU - Dutchuk, Mike
AU - Takahashi, Naoko
AU - Reddy, Ashok
AU - Hendricks, Jerry D.
AU - Williams, David E.
N1 - Funding Information:
The authors wish to thank Dan Arbogast of the Marine/Freshwater Biomedical Sciences Center for microinjection of trout embryos and Ted Will and Sheila Cleveland for preparation of diets and care and feeding of the fish. We thank Drs. Lawrence Curtis and Donald Buhler of the Marine/Freshwater Biomedical Sciences Center for their comments on this manuscript. We wish to acknowledge the Cancer Research Program of the National Cancer Institute, Division of Cancer Cause and Prevention, for making available [3H]anti-BPDE and [‘4C](+)-BP-7,8-DHD. We also express our gratitude to Dr. Lawrence Marnett, Center in Molecular Toxicology, Vanderbilt University, for providing the syn-and anti-BPDE-DNA standards. This work was supported by NIH grants ES03850 and ES04766. This manuscript was issued as Oregon State University Agricultural Experiment Station manuscript No. 10298.
PY - 1993/10/15
Y1 - 1993/10/15
N2 - Bioactivation in vivo of pure (+) 7S-trans-7,8-dihydrobenzo[a]pyrene-7,8-diol ((+) BP-7,8-DHD) was investigated in rainbow trout. Embryos, microinjected with 0.01-1.0 μg of [3H]-(-)-7S-trans-7,8-dihydrobenzo[a]-pyrene-7,8-diol-anti-9,10-epoxide ((-) anti-BPDE), exhibited a dose-dependent increase in DNA adduction. Subsequently, microinjection of trout embryos with [14C] (+) BP-7,8-DHD also demonstrated a dose-dependent increase in DNA adduction. To determine the relative contribution of P-450-dependent versus peroxidation-dependent epoxygenation of (+)-BP-7,8-DHD, trout embryos were co-injected with [14C]-(+)-BP-7,8-DHD and either β-naphthoflavone (BNF) (CYP1A1 inducer) or carbon tetrachloride (CCl4) (lipid peroxidation enhancer). Co-injection with BNF tended to enhance covalent binding to DNA, which was consistent with rapid induction of CYP1A1. Co-injection with CCl4, significantly increased covalent binding of [14C]-(+)-BP-7,8-DHD to DNA, suggesting a contribution from non-enzymic cooxidation. 32P-Postlabeling analysis of liver DNA adducts following i.p. injections of (+) BP-7,8-DHD did not detect appreciable amounts of (-) anti-BPDE-dG from juvenile trout fed control diets or diets containing hydrogen peroxide or BNF. On the contrary, BNF pre-feeding markedly enhanced the levels of an adduct which co-chromatographed with authentic (+) syn-BPDE-dG. These results confirm that trout are capable of metabolically activating BP-DHD to the ultimate carcinogen BPDE and that BNF stimulates CYP1A1-dependent epoxygenation, but peroxidation-dependent activation may not contribute significantly to the bioactivation of BP-7,8-DHD in vivo.
AB - Bioactivation in vivo of pure (+) 7S-trans-7,8-dihydrobenzo[a]pyrene-7,8-diol ((+) BP-7,8-DHD) was investigated in rainbow trout. Embryos, microinjected with 0.01-1.0 μg of [3H]-(-)-7S-trans-7,8-dihydrobenzo[a]-pyrene-7,8-diol-anti-9,10-epoxide ((-) anti-BPDE), exhibited a dose-dependent increase in DNA adduction. Subsequently, microinjection of trout embryos with [14C] (+) BP-7,8-DHD also demonstrated a dose-dependent increase in DNA adduction. To determine the relative contribution of P-450-dependent versus peroxidation-dependent epoxygenation of (+)-BP-7,8-DHD, trout embryos were co-injected with [14C]-(+)-BP-7,8-DHD and either β-naphthoflavone (BNF) (CYP1A1 inducer) or carbon tetrachloride (CCl4) (lipid peroxidation enhancer). Co-injection with BNF tended to enhance covalent binding to DNA, which was consistent with rapid induction of CYP1A1. Co-injection with CCl4, significantly increased covalent binding of [14C]-(+)-BP-7,8-DHD to DNA, suggesting a contribution from non-enzymic cooxidation. 32P-Postlabeling analysis of liver DNA adducts following i.p. injections of (+) BP-7,8-DHD did not detect appreciable amounts of (-) anti-BPDE-dG from juvenile trout fed control diets or diets containing hydrogen peroxide or BNF. On the contrary, BNF pre-feeding markedly enhanced the levels of an adduct which co-chromatographed with authentic (+) syn-BPDE-dG. These results confirm that trout are capable of metabolically activating BP-DHD to the ultimate carcinogen BPDE and that BNF stimulates CYP1A1-dependent epoxygenation, but peroxidation-dependent activation may not contribute significantly to the bioactivation of BP-7,8-DHD in vivo.
KW - Benzo[a]pyrene
KW - Carcinogenesis
KW - P-Post-labeling
KW - Trout
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U2 - 10.1016/0304-3835(93)90052-B
DO - 10.1016/0304-3835(93)90052-B
M3 - Article
C2 - 8287364
AN - SCOPUS:0027454255
SN - 0304-3835
VL - 74
SP - 111
EP - 117
JO - Cancer Letters
JF - Cancer Letters
IS - 1-2
ER -