Abstract
Although animal models have been used successfully to study metabolic activation and binding of carcinogens to DNA, only limited studies have been done in human systems. To circumvent the problems associated with the inaccessibility of human tissues and a lack of sensitive methods to detect DNA damage, we have investigated the capability of human peripheral blood lymphocytes in vitro to metabolize carcinogens to their DNA binding species by a 32P-labeled adduct assay. Freshly isolated lymphocytes were exposed at 37 degrees C for 18 hr to 4-aminobiphenyl, 2-aminofluorene, 2-anthramine, 2-acetylaminophenanthrene, benzidine, 1-nitropyrene, 1,2-benzanthracene, triphenylene, 7,12-dimethylbenz[a]anthracene, or benzo[a]pyrene at 30 microM each, compounds that are shown or suspected to be carcinogenic in experimental animals. Anthracene, pyrene, and perylene were included as noncarcinogenic controls. Our data indicate that all test carcinogens formed readily measurable levels of DNA adducts. Analysis of exposed DNAs by 32P-labeling after digestion and adduct enrichment showed exclusively or predominantly one major adduct for all test carcinogens, except for 2-anthramine, triphenylene, and 7,12-dimethylbenz[a]anthracene, which showed two or three adducts, in the range of 8-1500 amol/micrograms of DNA. No DNA binding was detected for the noncarcinogens. From 12 lymphocyte specimens studied thus far, significant interindividual variations were observed for 2-aminofluorene (62-fold), 7,12-dimethylbenz[a]anthracene (10-fold), benzidine (19-fold) and benzo[a]pyrene (18-fold) in their capacity to bind to the lymphocyte DNA. The lymphocyte system in combination with the 32P-adduct assay may prove to be an ultrasensitive means to determine interindividual variations in the ability to biotransform carcinogens.
Full text
PDF
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Beland F. A., Kadlubar F. F. Formation and persistence of arylamine DNA adducts in vivo. Environ Health Perspect. 1985 Oct;62:19–30. doi: 10.1289/ehp.856219. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gupta R. C., Dighe N. R. Formation and removal of DNA adducts in rat liver treated with N-hydroxy derivatives of 2-acetylaminofluorene, 4-acetylaminobiphenyl, and 2-acetylaminophenanthrene. Carcinogenesis. 1984 Mar;5(3):343–349. doi: 10.1093/carcin/5.3.343. [DOI] [PubMed] [Google Scholar]
- Gupta R. C. Enhanced sensitivity of 32P-postlabeling analysis of aromatic carcinogen:DNA adducts. Cancer Res. 1985 Nov;45(11 Pt 2):5656–5662. [PubMed] [Google Scholar]
- Gupta R. C. Nonrandom binding of the carcinogen N-hydroxy-2-acetylaminofluorene to repetitive sequences of rat liver DNA in vivo. Proc Natl Acad Sci U S A. 1984 Nov;81(22):6943–6947. doi: 10.1073/pnas.81.22.6943. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gupta R. C., Reddy M. V., Randerath K. 32P-postlabeling analysis of non-radioactive aromatic carcinogen--DNA adducts. Carcinogenesis. 1982;3(9):1081–1092. doi: 10.1093/carcin/3.9.1081. [DOI] [PubMed] [Google Scholar]
- Natarajan A. T., Obe G. Screening of human populations for mutations induced by environmental pollutants: use of human lymphocyte system. Ecotoxicol Environ Saf. 1980 Dec;4(4):468–481. doi: 10.1016/0147-6513(80)90049-4. [DOI] [PubMed] [Google Scholar]
- Oesch F., Aulmann W., Platt K. L., Doerjer G. Individual differences in DNA repair capacities in man. Arch Toxicol Suppl. 1987;10:172–179. doi: 10.1007/978-3-642-71617-1_16. [DOI] [PubMed] [Google Scholar]
- Sharma S., Mehta S., Morgan J., Maizel A. Molecular cloning and expression of a human B-cell growth factor gene in Escherichia coli. Science. 1987 Mar 20;235(4795):1489–1492. doi: 10.1126/science.3547651. [DOI] [PubMed] [Google Scholar]
- Waters M., Allen J., Doerr C., Erexson G., Hasselblad V., Huang S., Kligerman A., Moore M., Nesnow S., Stead A. A preliminary investigation of the parallelogram concept in genetic monitoring and risk estimation. Prog Clin Biol Res. 1986;207:203–215. [PubMed] [Google Scholar]
- Whitlock J. P., Jr, Cooper H. L., Gelboin V. H. Aryl hydrocarbon (benzopyrene) hydroxylase is stimulated in human lymphocytes by mitogens and benz(a)anthracene. Science. 1972 Aug 18;177(4049):618–619. doi: 10.1126/science.177.4049.618. [DOI] [PubMed] [Google Scholar]