Summary
The influence of four liver tumor promoters and two structurally related chemicals (non-promoters) on survival of normal adult rat hepatocytes was examined in primary culture. Of the four promoters, butylated hydroxytoluene (BHT), dichlorodiphenyltrichloroethane (DDT), barbital sodium and phenobarbital sodium, only phenobarbital efficiently prolonged the hepatocyte survival and maintained the morphological features of the cells. Both BHT and DDT were toxic to hepatocytes within the dose ranges tested. Barbital was also ineffective for maintenance of primary cultured hepatocytes but not toxic to the cells. Of the two non-promoters, barbituric acid and amobarbital, barbituric acid also showed no maintenance effect on hepatocytes. However, amobarbital resembled phenobarbital in its effect on the maintenance of hepatocytes in primary culture. DNA synthesis of primary cultured hepatocytes was severely suppressed by phenobarbital in a dose-dependent manner. The results clearly show that the ability to support long-term survival of primary cultured hepatocytes is a common property of some barbiturates but not of liver tumor promoters, and that the maintenance of hepatocytes by phenobarbital is not due to a counterbalance of stimulated proliferation and death of the cells.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- BHT:
-
butylated hydroxytoluene
- DDT:
-
dichlorodiphenyltrichloroethane
- EDTA:
-
ethylenediaminetetraacetic acid disodium
References
Barbason H, Rassenfosse C, Betz EH (1983) Promotion mechanism of phenobarbital and partial hepatectomy in DENA hepatocarcinogenesis cell kinetics effect. Br J Cancer 47:517–525
Berry MN, Friend DS (1969) High-yield preparation of isolated rat liver parenchymal cells. A biochemical and fine structural study. J Cell Biol 43:506–520
Bissell DM, Hammaker LE, Meyer UA (1973) Parenchymal cells from adult rat liver in nonproliferating monolayer culture. I. Functional study. J Cell Biol 59:722–734
Bonney RJ, Becker JE, Walker PR, Potter VR (1974) Primary monolayer cultures of adult rat liver parenchymal cells suitable for study of the regulation of enzyme synthesis. In Vitro 9:399–413
Diwan BA, Rice JM, Ohshima M, Ward JM, Dove LF (1985) Comparative tumor-promoting activities of phenobarbital, amobarbital, barbital sodium, and barbituric acid on livers and other organs of male F344/NCr rats following initiation withN-nitrosodiethylamine. J Natl Cancer Inst 74:509–516
Guillouzo A, Guguen-Guillouzo C, Boisnard M, Bourel M, Benhamou JP (1978) Smooth endoplasmic reticulum proliferation and increased cell multiplication in cultured hepatocytes of the newborn rat in the presence of phenobarbital. Exp Mol Pathol 28:1–9
Japundzic M, Knezevic B, Djordjevic-Camba V, Japundzic I (1967) The influence of phenobarbital-Na on the mitotic activity of parenchymal liver cells during rat liver regeneration. Exp Cell Res 48:163–167
Kitagawa T, Watanabe R, Kayano T, Sugano H (1980) In vitro carcinogenesis of hepatocytes obtained from acetylaminofluorenetreated rat liver and promotion of their growth by phenobarbital. Gann 71:747–754
Lee G-H, Sawada N, Mochizuki Y, Nomura K, Kitagawa T (1989) Immortal epithelial cells of normal C3H mouse liver in culture: possible precursor populations for spontaneous hepatocellular carcinoma. Cancer Res 49:403–409
Miyazaki M (1977) Primary culture of adult rat liver cells. I. Preparation of isolated cells from trypsin-perfused liver of adult rat. Acta Med Okayama 31:351–360
Miyazaki M, Tsunashima M, Wahid S, Miyano K, Sato J (1984) Comparison of cytologic and biochemical properties between liver cells isolated by trypsin perfusion and those isolated by collagenase perfusion. Res Exp Med 184:191–204
Miyazaki M, Handa Y, Oda M, Yabe T, Miyano K, Sato J (1985 a) Long-term survival of functional hepatocytes from adult rat in the presence of phenobarbital in primary culture. Exp Cell Res 159:176–190
Miyazaki M, Handa Y, Sato J (1985 b) Effect of phenobarbital on adult rat liver cells treated with 3′-mefhyl-4-dimethylaminoazobenzene in primary culture. J Cancer Res Clin Oncol 110:191–195
Miyazaki M, Handa Y, Suzuki Y, Sato J (1986) Promotion of 3′-methyl-4-dimethylaminoazobenzene-initiated adult rat liver cells to malignant state by phenobarbital in culture. Int J Cancer 37:769–773
Moore MA, Kitagawa T (1986) Hepatocarcinogenesis in the rat: the effect of promoters and carcinogens in vivo and in vitro. Int Rev Cytol 101:125–173
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
Peraino C, Fry RJM, Staffeldt E, Kisieleski WE (1973) Effects of. varying the exposure to phenobarbital on its enhancement of 2-acetylaminofluorene-induced hepatic tumorigenesis in the rat. Cancer Res 33:2701–2705
Peraino C, Fry RJM, Staffeldt E, Christopher JP (1975) Comparative enhancing effects of phenobarbital, amobarbital, diphenylhydantoin, and dichlorodiphenyltrichloroethane on 2-acetylaminofluorene-induced hepatic tumorigenesis in the rat. Cancer Res 35:2884–2890
Peraino C, Fry RJM, Staffeldt E, Christopher JP (1977) Enhancing effects of phenobarbitone and butylated hydroxytoluene on 2-acetylaminofluorene-induced hepatic tumorigenesis in the rat. Food Cosmet Toxicol 15:93–96
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Miyazaki, M., Bai, L. & Sato, J. Influence of liver tumor promoters and structurally related chemicals on survival of normal adult rat hepatocytes in primary culture. J Cancer Res Clin Oncol 116, 259–263 (1990). https://doi.org/10.1007/BF01612900
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01612900