Summary
The effect of phenobarbital (PB) on liver cells treated with 3′-methyl-4-dimethylaminoazobenzene (3′-Me-DAB) was studied using primary cultures of normal adult rat liver cells. Following a 1-day attachment period, primary liver cell cultures were treated with 0.24 mM 3′-Me-DAB for 6 days, and then treated with or without PB at 0.75, 1.5 and 3 mM for 19 days. Similarly, control cultures were treated with 0.5% dimethylsulfoxide (DMSO), a solvent for 3′-Me-DAB, for 6 days, and then treated with or without PB in the same way. Each treatment was done on 8 cultures. Chromosome analysis and cytochemical assay for gamma-glutamyltranspeptidase (GGT) activity were carried out on the carcinogen-treated and control cultures between 1 and 2 months after initiation of primary culture. Chromosomal abnormalities were detected in 23 of 32 carcinogen-treated cultures and also in 2 of 28 control cultures tested. However, GGT positive cells were detected only in the carcinogen-treated cultures at a frequency of 22/32. Of the 23 carcinogen-treated cultures with chromosomal abnormalities, 18 contained GGT positive cells. These results show a good correlation between chromosomal abnormality and acquisition of GGT activity at culture dish level. Furthermore, in the carcinogen-treated cultures, PB treatment caused a dose-dependent increase in the number of GGT positive cultures and in the percentage of GGT positive cells in each culture, and also caused a dose-dependent increase in the number of cultures with chromosomal abnormalities.
Similar content being viewed by others
References
Albert Z, Orlowski M, Szewezuk A (1961) Histochemical demonstration of gamma-glutamyl transpeptidase. Nature 191:767–768
Berenblum I, Haran-Ghera N (1955) The significance of the sequence of initiating and promoting actions in the process of skin carcinogenesis in mouse. Br J Cancer 9:268–271
Colburn NH, Former BF, Nelson KA, Yuspa SH (1979) Tumor promoter induces anchorage independence irreversibly. Nature 281:589–591
Hirota N, Williams GM (1979a) The sensitivity and heterogeneity of histochemical markers for altered foci involved in liver carcinogenesis. Am J Pathol 95:317–328
Hirota N, Williams GM (1979b) Persistence and growth of rat liver neoplastic nodules following cessation of carcinogen exposure. J Natl Cancer Inst 63:1257–1265
Kitagawa T, Sugano H (1978) Enhancing effect of phenobarbital on the development of enzyme-altered islands and hepatocellular carcinomas initiated by 3′-methyl-4-(dimethylamino)azobenzene or dietnylnitrosamine. Gann 69:679–687
Laishes BA, Ogawa K, Roberts E, Farber E (1978) Gamma-glutamyl transpeptidase: a positive marker for cultured rat liver cells derived from putative premalignant and malignant lesions. J Natl Cancer Inst 60:1009–1016
Lasne C, Gentil A, Chouroulinkov I (1974) Two stage malignant transformation of rat fibroblasts in tissue culture. Nature 247:490–491
Miyazyki M (1978) Primary culture of adult rat liver cells. II. Cytological and biochemical properties of primary cultured cells. Acta Med Okayama 32:11–22
Miyazaki M, Wahid S, Matsuura K, Tsunashima M, Nakabayashi H, Miyano K, Sato J (1982a) Effect of 3′-methyl-4-dimethylaminoazobenzene on liver cells from adult rat in primary culture. Res Exp Med 181:27–38
Miyazaki M, Wahid S, Miyano K, Sato J (1982b) Selective growth of epithelial-like clear cells from adult rat liver by short-term exposure to glucocorticoids in primary culture. Res Exp Med 181:189–196
Miyazaki M, Wahid S, Miyano K, Yabe T, Sato J (1983) Expression of gamma-glutamyltranspeptidase in cultures of spontaneously and chemically transformed rat liver cells. Int J Cancer 32:373–377
Miyazaki M, Tsunashima M, Wahid S, Miyano K, Sato J (1984) Comparison of cytologic and biochemical properties between liver cells isolated from adult rats by trypsin perfusion and those by collagenase perfusion. Res Exp Med 184:191–204
Mondal S, Brankow DW, Heidelberger C (1976) Two-stage chemical oncogenesis in cultures of C3H/10T1/2 cells. Cancer Res 36:2254–2260
Peraino C, Fry RJM, Staffeldt E (1971) Reduction and enhancement by phenobarbital of hepatocarcinogenesis induced in the rat by 2-acetylaminofluorene. Cancer Res 31:1506–1512
Rothfels KH, Siminovitch L (1958) An air drying technique for flattening chromosomes in mammalian cells grown in vitro. Stain Technol 33:73–77
Weisburger JH, Madison RM, Ward JM, Viguera C, Weisburger EK (1975) Modification of diethylnitrosamine liver carcinogenesis with phenobarbital but not with immunosuppression. J Natl Cancer Inst 54:1185–1188
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Miyazaki, M., Handa, Y. & Sato, J. Effect of phenobarbital on adult rat liver cells treated with 3′-methyl-4-dimethylaminoazobenzene in primary culture. J Cancer Res Clin Oncol 110, 191–195 (1985). https://doi.org/10.1007/BF00399272
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00399272