Estrogen-induced endogenous DNA adduction: possible mechanism of hormonal cancer.

Liehr, Joachim G.; Avitts, Tommie A.; Randerath, Erika; Randerath, Kurt

doi:10.1073/pnas.83.14.5301

Cited by 123 publications

(56 citation statements)

References 27 publications

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“…A general and sensitive approach for the measurement of DNA lesions formed with nonradioactive carcinogens has been described (12)(13)(14)(15), in which normal and adducted nucleotides, generated by nuclease digests of DNA modified in vivo or in vitro with a compound of interest, are labeled with 32P and detected and quantitated after TLC. The 32P-approach has been applied to approximately 100 test chemicals, comprising arylamines and derivatives, azo compounds, nitroaromatics, polycyclic aromatic hydrocarbons, and methylating agents (14), as well as mycotoxins (16), heterocyclic polycyclic aromatics (17), alkenylbenzene derivatives (18,19), and estrogens (20). With each carcinogen tested, 32P-labeled DNA adducts were readily detected, showing the potential use of the 32P-assay as a shortterm in vivo test for screening genotoxic chemicals.…”

Section: Introductionmentioning

confidence: 99%

32P-postlabeling assay for carcinogen-DNA adducts: nuclease P1-mediated enhancement of its sensitivity and applications.

Reddy

Randerath

1987

Environ Health Perspect

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Exceedingly sensitive assays are required for the detection of DNA adducts formed in humans exposed to low levels of environmental genotoxicants and therapeutic drugs. A 3P-postlabeling procedure for detection and quantitation of aromatic carcinogen-DNA lesions with a sensitivity limit of 1 adduct in 107 to 108 nucleotides has been described previously. In the standard procedure, DNA is enzymatically digested to 3'-phosphorylated normal and adducted mononucleotides, which are 32P-labeled at their 5'-hydroxyl groups by T4 polynucleotide kinase-catalyzed [32P]phosphate transfer from [y-32P]ATP. 3P-labeled derivatives are resolved by TLC, detected by autoradiography, and quantitated by counting. This assay has been recently utilized for the determination and partial characterization of DNA adducts formed in somatic and reproductive tissues of rats given the clinically used anticancer drug, mitomycin C. The drug exhibits similar levels of covalent binding to DNA in most tissues. Further studies have revealed that adducted nucleotides are primarily guanine derivatives that are resistant to 3'-dephosphorylation by Penicillium citrinum nuclease Pl. The latter observation has been utilized to enhance the 32P-assay's sensitivity to 1 adduct in 1010 nucleotides for a 10-R±g DNA sample by postincubation of DNA digests with nuclease P1 before 32P-labeling. The enzyme dephosphorylates the normal nucleotides but not most aromatic and bulky nonaromatic adducts, so that only the latter serve as substrates for the kinase-catalyzed labeling reaction. The new assay has also shown utility in the analysis of very low levels of age-and tissue-related DNA modifications, which might arise from dietary or endogenous compounds, in untreated rats and in humans.

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Section: Introductionmentioning

confidence: 99%

32P-postlabeling assay for carcinogen-DNA adducts: nuclease P1-mediated enhancement of its sensitivity and applications.

Reddy

Randerath

1987

Environ Health Perspect

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“…However, it is not clear whether this adduct is derived from DNA-binding B[a]P metabolites or whether it is an indirect DNA modification that does not contain the B[a]P moiety. It is known that indirect DNA adducts with unknown chemical identity could be generated during metabolism of carcinogens and estrogens [Liehr et al, 1986, Marnett et al, 1993. To distinguish whether Spot 1 is directly derived from B[a]P or is an endogenous DNA adduct formed as a consequence of B[a]P exposure, we compared it with adduct profiles de- Most of these compounds induced DNA adduct formation but none of these adducts is in a location similar to that of Spot 1 under our chromatography conditions (films not shown).…”

Section: Resultsmentioning

confidence: 99%

Characterization of a major aromatic DNA adduct detected in human breast tissues

Wang

Firozi

et al. 2002

Environ and Mol Mutagen

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A bulky DNA adduct (Spot 1) was previously detected in normal adjacent breast tissues of 41% (36/87) of women with breast cancer and in none (0/29) of the noncancer controls by 32 P-postlabeling. To characterize this adduct, it was chromatographically compared with DNA adduct profiles generated in several in vitro and in vivo experimental systems. First, MCF-7 cells were exposed to a number of chemical carcino- [a,h]anthracene, 3-methylcholanthrene, and 2-amino-1-methyl-6-phenylimidazo [4,5-b]pyridine. Spot 1 was detected as a minor adduct in cells treated with B[a]P but not other compounds. Second, to determine whether Spot 1 is derived from lipid peroxidation products or estrogen metabolites, it was compared with adduct profiles of cells or DNAs exposed to 17␤-estradiol, 4-hydroxy estradiol, 4-hydroxynonenal, or oxidized oat oil. Spot 1 was not detectable in these samples. In addition, Spot 1 did not comigrate with the 1,N 2 -ethenodeoxyguanosine adduct standard. Third, to explore the mechanism of Spot 1 formation, it was compared with adduct profiles detected in DNA or mononucleotides reacted with BPDE, 1-OH-7,8-dihydrodiol of B[a]P, and 3-OH-7,8-dihydrodiol of B[a]P as well as in rats orally treated with B[a]P. Spot 1 comigrated with a minor adduct in BPDEtreated DNA during anion exchange rechromatography but these two adducts were separated by partition chromatography. Spot 1 also behaved in a manner that was very similar to that of the polar B[a]P adducts detected in rat liver, but the two adducts were separated by HPLC. Fourth, Spot 1 was compared with CD1 mice exposed to 7H-benzo[c]fluorene (B[c]F). Spot 1 from some patients comigrated with a major adduct induced by B[c]F. Finally, we found that the presence of Spot 1 in human breast tissues was not related to smoking status but, rather, with CYP1A1 MspI polymorphism. The CYP1A1 mutant carriers had a significantly higher frequency of this adduct than did the wild-type genotypes. Furthermore, individuals with Spot 1 had a significantly higher staining intensity for BPDE-PAH adducts in their tissue sections than those without it. These results demonstrate that this major bulky DNA adduct detected in human breast tissues is related to PAH exposure. Environ. Mol. Mutagen. 39:193-200, 2002.

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“…DNA adducts were readily detectable (20). As shown in Figure 6, when 32p-labeled digests were chromatographed according to -, -MP FIGURE 5.…”

Section: Analysis Of Adducts From Benzene and Its Derivativesmentioning

confidence: 99%

A Method for In Vitro Culture of Rat Zymbal Gland: Use in Mechanistic Studies of Benzene Carcinogenesis in Combination with 32 P-Postlabeling

Reddy¹,

Blackburn²,

Irwin³

et al. 1989

Environmental Health Perspectives

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Zymbal glands were excised bilaterally from the ear ducts of female Sprague-Dawley rats (three/group), minced into approximately four fragments per gland, and transferred into a microtiter plate containing 1.5 mL per well of Waymouth's tissue culture medium supplemented with fetal calf serum, hydrocortisone, insulin, and gentamicin. After addition of a test compound or solvent vehicle, plates were incubated for 6, 24, 48, or 96 hr at 37°C in a humidified atmosphere of 5% CO2 in air. Tissue in culture for 6 hr was histologically indistinguishable from the freshly excised tissue, while that in culture for 24, 48, and 96 hr showed a progressive deterioration often with necrosis and/or squamous metaplasia. More pronounced deterioration was noted in samples treated with 750 or 1500 pg/mL of benzene. Using a nuclease Pi-enhanced 32P-postlabeling assay, aromatic DNA adducts were detected in cultured Zymbal glands exposed for 48 hr to benzene and its derivatives, as well as to 7,12-dimethylbenzanthracene (DMBA) and 2-acetylaminofluorene (AAF). Benzene produced very low levels of adducts (0.5 adducts per 109 nucleotides), whereas its congeners produced relatively high levels of adducts (50-2000 lesions per 109 nucleotides), which decreased in the order benzoquinone > hydroquinone > phenol > benzenetriol > catechol. Each adduct profile overall was characteristic for the compound studied, suggesting the formation of compound-specific electrophiles. AAF and DMBA adducts were identical to those formed in vivo in animals. Our results show that the Zymbal glands are capable of metabolizing different carcinogens to DNA-reactive intermediates, a process that may be causally associated with tumor formation in vivo in this organ.

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Estrogen-induced endogenous DNA adduction: possible mechanism of hormonal cancer.

Cited by 123 publications

References 27 publications

32P-postlabeling assay for carcinogen-DNA adducts: nuclease P1-mediated enhancement of its sensitivity and applications.

32P-postlabeling assay for carcinogen-DNA adducts: nuclease P1-mediated enhancement of its sensitivity and applications.

Characterization of a major aromatic DNA adduct detected in human breast tissues

A Method for In Vitro Culture of Rat Zymbal Gland: Use in Mechanistic Studies of Benzene Carcinogenesis in Combination with 32 P-Postlabeling

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