Immunocytochemical visualization of DNA adducts in mouse tissues and human white blood cells following treatment with benzo[a]pyrene or its diol epoxide. A quantitative approach
| Autor: | Michel J.X. Hillebrand, F.J. van Schooten, E. Kriek, E. Schere, L. Den Engelse |
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| Rok vydání: | 1991 |
| Předmět: |
Male
Cancer Research DNA repair 7 8-Dihydro-7 8-dihydroxybenzo(a)pyrene 9 10-oxide Enzyme-Linked Immunosorbent Assay Mice Inbred Strains Kidney Dihydroxydihydrobenzopyrenes Adduct DNA Adducts Mice chemistry.chemical_compound DNA adduct Benzo(a)pyrene Leukocytes Animals Lung Cells Cultured Carcinogen Dose-Response Relationship Drug Myocardium DNA General Medicine Immunohistochemistry chemistry Biochemistry Pyrene Female Thymidine Phosphorus Radioisotopes Densitometry |
| Zdroj: | Carcinogenesis. 12:427-433 |
| ISSN: | 1460-2180 0143-3334 |
| DOI: | 10.1093/carcin/12.3.427 |
| Popis: | The formation and stability of benzo[a]pyrene DNA adducts were studied in tissues of BALB/c mice exposed to benzo[a]pyrene (B[a]P). The DNA adducts were visualized with an immunocytochemical peroxidase staining technique using an antiserum specific for the major B[a]P-derived adduct in DNA [(+/-)trans-7,8-dihydroxy-anti-9,10-epoxy-7,8,9,10- tetrahydrobenzo[a]pyrene (BPDE-N2-dG)]. The nuclear staining density was measured by microdensitometry. When mice were treated with an increasing dose of B[a]P the nuclear staining increased in the tissues studied (lung, heart and kidney). A linear relationship was found between the immunocytochemical nuclear staining signal and the actual DNA adduct level in the lung as measured by 32P-postlabeling. Maximum adduct formation was found 5 days after a single i.p. injection of B[a]P. Adduct levels decreased gradually after 7 days, but even after 61 days a slight specific staining was still present, suggesting that not all adducts had disappeared at that time. As judged from the disappearance of [3H]thymidine from prelabeled DNA the loss of adducts from the lung was not a result of DNA repair but one of cell turnover. In human white blood cells B[a]P-derived adducts could be detected after in vitro incubation with the reactive metabolite of B[a]P (BPDE). Dose-response studies demonstrated a positive relationship between BPDE-DNA adduct formation, the immunocytochemical staining signal and the BPDE concentration in the culture medium. |
| Databáze: | OpenAIRE |
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