HPLC separation of 32P-postlabelled DNA adducts formed from dibenz[a,h]anthracene in skin

Lecoq, Sophie; Pfau, Wolfgang; Grover, Philip L.; Phillips, David H.

doi:10.1016/0009-2797(92)90060-x

Cited by 12 publications

(5 citation statements)

References 21 publications

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“…The existence of highly polar PAH-DNA adducts was recently shown for another isomer of DBA, DB[a,h]A (41), as well as for several other PAH (42,43). The existence of highly polar DNA adducts in epidermal DNA samples from DB[aj]A-treated mice was verified through use of a 32P-postlabeling technique coupled with HPLC.…”

Section: Discussionmentioning

confidence: 94%

Covalent DNA Adducts Formed in Mouse Epidermis from Dibenz[a,j]anthracene: Evidence for the Formation of Polar Adducts

Baer‐Dubowska

Nair

Cortez

et al. 1995

Chem. Res. Toxicol.

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The formation of deoxyribonucleoside adducts in mouse epidermis has been examined following topical application of [3H]dibenz[a,j]anthracene (DB[a,j]A) or by 32P-postlabeling following topical application of unlabeled DB[a,j]A, DB[a,j]A trans-3,4-diol or the anti- or syn-3,4-diol 1,2-epoxides. A single topical application of [3H]DB[a,j]A at a dose of 400 nmol per mouse led to the formation of 11 detectable covalent DNA adducts. Seven of these DNA adducts were tentatively identified based on cochromatography with marker adducts using high-pressure liquid chromatography (HPLC). The presence of both deoxyguanosine (dGuo) as well as deoxyadenosine (dAdo) adducts formed from bay-region anti- and syn-3,4-diol 1,2-epoxides of DB[a,j]A was revealed. The major bay-region diol epoxide DNA adduct formed in mouse epidermis following topical application of [3H]DB[a,j]A was tentatively identified as the (4R,3S)-diol (2S,1R)-epoxide bound through trans addition of the exocyclic amino group of dGuo, although substantial amounts of the corresponding dAdo adduct were also detected. In addition, a K-region 5,6-oxide-dAdo adduct was tentatively identified in HPLC chromatograms based on cochromatography with an authentic marker adduct. 32P-Postlabeling analysis of DB[a,j]A-DNA adducts formed in mouse epidermis after topical application of unlabeled compound confirmed the presence of bay-region diol epoxide DNA adducts similar to those observed after application of [3H]DB[a,j]A. However, 32P-postlabeling analysis also revealed the presence of more polar covalent DNA adducts in epidermal DNA samples from DB[a,j]A-treated mice. These more polar DNA adducts represented a significant proportion of the 32P-labeled material recovered in HPLC chromatograms. While the exact nature of these adducts remains unknown at present, they had retention times identical to polar DNA adducts formed following topical application of DB[a,j]A trans-3,4-diol and may represent bis-dihydrodiol epoxide DNA adducts. The present results indicate that a rather broad spectrum of DNA adducts arises following topical application of DB[a,j]A to mouse epidermis.

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Section: Discussionmentioning

confidence: 94%

Covalent DNA Adducts Formed in Mouse Epidermis from Dibenz[a,j]anthracene: Evidence for the Formation of Polar Adducts

Baer‐Dubowska

Nair

Cortez

et al. 1995

Chem. Res. Toxicol.

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“…This symmetrical compound can form not only the bay-region 3,4-diol-1,2-epoxide, but also the 3,4:10,11-bis-diol-1,2-epoxide (Carmichael et al, 1993; Fuchs et al, 1993a, 1993b). When the metabolism, mutagenicity, carcinogenicity and formation of DNA adducts of this compound were studied, all of the results indicated that it is activated via the diol epoxide pathway (Fuchs et al, 1993a, 1993b; Lecoq et al, 1991, 1992; Platt, et al, 1990). One study suggests that the 3,4:10,11-bis-diol-1,2-epoxide is the most important for the carcinogenicity of this compound (Carmichael et al, 1993).…”

Section: Ultimate Carcinogenic Metabolites Of Aromatic Hydrocarbonsmentioning

confidence: 99%

The molecular etiology and prevention of estrogen-initiated cancers

Cavalieri

Rogan

2014

Molecular Aspects of Medicine

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“…Therefore, it will be interesting to determine the stereoselectivity in the metabolic conversion of enantiomeric DBA-3,4-dihydrodiol and to eluciate the role of the stereoisomers of the bisdihydrodiols in the genotoxicity of DBA. The relevance of the metabolic pathways elucidated in this study for the genotoxic activity of DBA has been furthermore ascertained by determination of the DNA binding in vitro (36,38,39) and in vivo (40)(41)(42). Thus the postulated reactivity of the ultimate mutagens of the three bisdihydrodiols was confirmed by the extent of DNA binding in mouse skin (42), which rose from no detectable binding in the case of DBA-3,4:12,13-bisdihydrodiol to weak binding in the case of DBA-3,4:8,9-bisdihydrodiol, reaching its highest value in the case of DBA-3,4:10,11bisdihydrodiol.…”

Section: Resultsmentioning

confidence: 99%

“…In both metabolizing systems DNA adducts based on bay-region dihydrodiol oxides were detected (38)(39)(40) and could be characterized ( 39), yet, especially in vivo, their amount was surpassed by more polar adducts (40). Remarkably, this could be shown not only with mouse skin (40) but also with human skin in culture (41). Finally, it was shown in vitro (36) and in vivo (42) that these polar DNA adducts are formed from DBA-3,4:10,11-bisdihydrodiol through further metabolic activation.…”

Section: Resultsmentioning

confidence: 99%

Bisdihydrodiols, rather than dihydrodiol oxides, are the principal microsomal metabolites of tumorigenic trans-3,4-dihydroxy-3,4-dihydrodibenz[a,h]anthracene

Platt¹,

Schollmeier²

1994

Chem. Res. Toxicol.

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Several studies on metabolism and biological activity of tumorigenic dibenz[a,h]anthracene (DBA) and its derivatives have led to the conclusion that the M-region dihydrodiol, trans-3,4-dihydroxy-3,4-dihydro-DBA (DBA-3,4-dihydrodiol), is the precursor of the ultimate mutagenic and tumorigenic metabolite of DBA with the presumed structure of a bay-region dihydrodiol oxide. Incubations of DBA-3,4-dihydrodiol (50 microM) with the microsomal hepatic fraction of Sprague-Dawley rats pretreated with Aroclor 1254 yielded more than 13 metabolites upon separation by HPLC. anti-3,4-Dihydroxy-1,2-epoxy-1,2,3,4-tetrahydro-DBA [0.27 nmol/(nmol of P450.15 min)] could be identified for the first time by UV spectroscopy, by cochromatography with the synthetic reference compound, and by its nonenzymatic hydrolysis to r-1,t-2,t-3,c-4-tetrahydroxy-1,2,3,4-tetrahydro-DBA, while firm evidence for the presence of the diastereomeric syn-dihydrodiol oxide was not obtained. Major microsomal metabolites of the M-region dihydrodiol were however three bisdihydrodiols: trans,trans-3,4:8,9-tetrahydroxy-3,4,8,9-tetrahydro-DBA [0.32 nmol/(nmol of P450.15 min)], trans,trans-3,4:10,11-tetrahydroxy-3,4,-10,11-tetrahydro-DBA [DBA-3,4:10,11-bisdihydrodiol; 1.44 nmol/(nmol of P450.15 min)], and trans,trans-3,4:12,13-tetrahydroxy-3,4,12,13-tetrahydro-DBA [0.70 nmol/(nmol of P450.15 min)], whose structures were verified by UV and mass spectrometry as well as cochromatography with synthetic reference compounds and by the observation that they were not formed when epoxide hydrolase was inhibited (1,1,1-trichloro-2-propene oxide, 1 mM).(ABSTRACT TRUNCATED AT 250 WORDS)

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HPLC separation of 32P-postlabelled DNA adducts formed from dibenz[a,h]anthracene in skin

Cited by 12 publications

References 21 publications

Covalent DNA Adducts Formed in Mouse Epidermis from Dibenz[a,j]anthracene: Evidence for the Formation of Polar Adducts

Covalent DNA Adducts Formed in Mouse Epidermis from Dibenz[a,j]anthracene: Evidence for the Formation of Polar Adducts

The molecular etiology and prevention of estrogen-initiated cancers

Bisdihydrodiols, rather than dihydrodiol oxides, are the principal microsomal metabolites of tumorigenic trans-3,4-dihydroxy-3,4-dihydrodibenz[a,h]anthracene

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