The genotoxic activity of benzidine was studied in two cell lines derived from rat (H4) and human (HepG2) hepatomas which have been shown to be capable of activating certain promutagens. The responses were compared to results in two lung-derived fibroblast lines (IMR-90 and V79) which appear to have little or no metabolizing capability. Benzidine was found to induce sister chromatid exchanges in the two liver-derived cell lines in a dose-dependent fashion but failed to induce sister chromatid exchanges in the fibroblast lines. Since one proposed pathway for benzidine activation involves prostaglandin-mediated metabolism, we tested the effect of pretreatment with indomethacin, an inhibitor of this metabolic pathway. Indomethacin was highly effective in inhibiting benzidine-induced sister chromatid exchanges in both H4 and HepG2 cells. These results suggest that some DNA damage may occur in the livers of fast acetylating species such as the rat without prior N-acetylation and that some amount of DNA damage may occur in the livers of slow acetylating species, even when the liver is not the target organ for carcinogenesis.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- RI:
-
replication index
- SCE:
-
sister chromatid exchanges
References
DEARFIELD, K.L., JACOBSON-KRAM, D., BROWN, N.A. and WILLIAMS, J.R. (1983). Evaluation of a human hepatoma cell line as a target cell line in genetic toxicology. Mutat. Res. 108:437–449.
GLOWINSKI, I.B., RADTKE, H.E. and WEBER, W. (1978). Genetic variation in N-acetylation of carcinogenic arylamines by human rabbit liver. Mol. Pharmacol. 14:940–949.
HALEY, T.J. (1975). Benzidine revisited: A review of the literature and problems associated with the use of benzidine and its congeners. Clin. Toxicol. 8:13–42.
HIRSCH, B., McGUE, M. and CERVENKA, J. (1984). Characterization of the distribution of sister chromatid exchange frequencies: Implications for research design. Human Genet. 65:280–286.
IARC MONOGRAPHS ON THE EVALUATION OF CARCINOGENIC RISK OF CHEMICALS TO MAN. (1982). Vol 29, International Agency for Research on Cancer, Lyon.
JOSEPHY, P.D., ELING, T.E. and MASON, R.P. (1983). Co-oxidation of benzidine by prostaglandin synthetase and comparison with the action of horseradish peroxidase. Jour. Biol. Chem. 258:5561–5569.
KENNELLY, J.C., BELAND, F.A., KADLUBAR, F.F. and MARTIN, C.N. (1984). Binding of N-acetylbenzidine and N,N′-diacetylbenzidine to hepatic DNA of rat and hamster in vivo and in vitro. Carcinogenesis 5:407–412.
KNOWLES, B., HOWE, C. and ADEN, D. (1980). Human hepatocellular carcinoma cell lines secrete the major plasma and hepatitis B surface antigen. Science 209:497–499.
LIN, M. and ALFI, O. (1976). Detection of sister chromatid exchanges by 4′-6-diamidino-2-phenylindole fluorescence. Chromosoma 57:129–135.
LOWER, G.M., NILSSON, T., NELSON, C.E., WOLF, H., GAMSKY, T.E. and BRYAN, G.T. (1979). N-acetyltransferase phenotype and risk in urinary cancer: Approaches in molecular epidemiology. Preliminary results in Sweden and Denmark. Environ. Health Perspect. 29:71–79.
MANCUSO, T.F. and EL ATTAR, A.A. (1967). Cohort study of workers exposed to beta-naphthylamine and benzidine. Jour. Occup. Med. 9:277–285.
MARNET, L.J., WLODAWER, P. and SAMUELSSON, B. (1975). Co-oxygenation of organic substrates by prostaglandin synthetase of sheep vesicular gland. Jour. Biol. Chem. 250:8510–8517.
MARTIN, C.N., BELAND, F.A., ROTH, R.W. and KADLUBAR, F.F. (1982). Covalent binding of benzidine and N-acetylbenzidine to DNA at the C-8 atom of deoxyguanosine in vivo and in vitro. Cancer Res. 42:2678–2696.
MORTON, K.C., KING, C.M. and BAETCKE, K.P. (1979). Metabolism of benzidine to N-hydroxyl-N, N′-diacetylbenzidine and subsequent nucleic acid binding and mutagenicity. Cancer Res. 39:3107–3113.
MORTON, K.C., KING, C.M., VAUGHT, J.B., WANG, C.Y., LEE, M.S. and MARNETT, L.J. (1983). Prostaglandin H synthetase-mediated reaction of carcinogenic arylamines with tRNA and homopolyribonucleotides. Biochem. Biophys. Res. Commun. 111:96–103.
NELSON, C.J., BAETCKE, F.P., FRITH, C.H., KODELL, R.L. and SCHIEFERSTEIN, G. (1982). The influence of sex, dose, time and cross on neoplasia in mice given benzidine dihydrochloride. Toxicol. Appl. Pharmacol. 64:171–186.
PHILLIPSON, C.E. and IOANNIDES, C. (1983). Activation of aromatic amines to mutagens by various animal species including man. Mutat. Res. 124:326–336.
PITOT, H.C., PERAINO, C., MORSE, P.A. and POTTER, V. (1964). Hepatomas in tissue culture compared with adapting in vivo. NCI Monograph 13:229–245.
PIUPKO, J.M., RADOMSKI, T., SANTELLA, R.M. and RADOMSKI, J.L. (1983). Organ, species and compound specificity in metabolic activation of primary aromatic amines. Jour. Natl. Cancer Instit. 70:1077–1080.
REUBER, M. (1961). A transplantable bile-secreting hepatocellular carcinoma in the rat. Jour. Natl. Cancer Instit. 26:891–897.
SCIARNI, L.J. and MEIGS, J.W. (1958). The biotransformation of benzidine (4,4′-diaminobiphenyl), an industrial carcinogen in the dog. IAMA Arch. Ind. Health 18:521–530.
SCIARNI, L.J. and MEIGS, J.W. (1961). The biotransformation of benzidine. 2. Studies in mouse and man. Arch. Environ. Health 2:423–433.
VESSELINOVITCH, L.D., RAO, K.V.N. and MIHAILOVICH, N. (1975). Factors modulating benzidine carcinogenicity bioassay. Cancer Res. 38:2814–2819.
WISE, R.W., ZENSER, T.V. and DAVIS, B.B. (1983). Prostaglandin H synthetase metabolism of the urinary bladder carcinogens benzidine and ANFT. Carcinogenesis 4:285–289.
WISE, R.W., ZENSER, T.V., KADLUBAR, F.F. and DAVIS, B.B. (1984). Metabolic activation of carcinogenic aromatic amines by dog bladder and kidney prostaglandin H synthetase. Cancer Res. 44:1893–1897.
ZENSER, T.V., MATTAMMAL, M.B., ARMECHT, H.J. and DAVIS, B.B. (1980). Benzidine binding to nucleic acids mediated by peroxidase activity of prostaglandin endoperoxidase synthetase. Cancer Res. 40:2839–2845.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Grady, M.K., Jacobson-Kram, D., Dearfield, K.L. et al. Induction of sister chromatid exchanges by benzidine in rat and human hepatoma cell lines and inhibition by indomethacin. Cell Biol Toxicol 2, 223–230 (1986). https://doi.org/10.1007/BF00122691
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00122691