Skip to main content

Toxic effects of 70-hydroxycholesterol on rat liver primary cultures, epithelial lines and co-cultures.

Cell Biology and Toxicology volume 5, pages 261–270 (1989)Cite this article

Abstract

The toxic effects of 7,8-hydroxycholesterol (7β-OHC) on cultures and co-cultures of rat hepatocytes, rat liver epithelial cell lines, and rat liver fibroblast lines were investigated. Hepatocytes in primary culture or co-cultured with proliferative epithelial cells, were not affected by the presence of 7,8-OHC at a concentration of 400µM over a period of 72 hours. In contrast, Proliferative cultures of liver epithelial cell lines and liver fibroblast lines were killed by 50 µM 7β-OHC within the first 24 hours. Established liver epithelial cells (hyperploid) were more sensitive to 7,8-OHC than the same line at early passages (diploid). When hepatocytes and liver epithelial cells were co-cultured and treated with 100 µM 7β-OHC only epithelial cells were lysed. A concentration of 50 µM 7β-OHC was toxic to co-cultures of liver epithelial cell and fibroblasts together. In a serum free medium, the cytotoxic concentration of 7β-OHC was lower than that in the serum-supplemented medium. Thus, liver epithelial cells cultured alone or co-cultured with hepatocytes were killed at 12.5 µM and 50 µM 7β-OHC, respectively. Finally, cholesterol concentrations four fold that of 7β-OHC antagonized the lethal effects of 7β-OHC in the serum free medium.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

Abbreviations

7β-OHC:

7β-hydroxycholesterol

SFM:

serum-free medium

SSM:

serum-supplemented medium

References

  • Cheng, K.P., Nagano, H., Luu, B., Ourisson, G. and Beck, J.P. (1977). Chemistry and biochemistry of chinese drugs. I. Sterol derivatives cytotoxic to hepatoma cells isolated from the drug Bombyx cum Botryte. J. Chem. Res.217:2511–2521.

    Google Scholar 

  • Chessebeuf, M., Olsson, A., Bournot, P., Desgres, J., Guiguet, M., Maume, G., Perissel, B. and Padieu, P. (1975). Retention and loss of certain enzymes in various primary cultures and cell lines of normal rat liver. In: Gene Expression and Carcinogenesis in Cultured Liver (L.F. Gerschenson and E.B. Thompson, eds), pp 94–117, Academic Press, New York.

    Google Scholar 

  • Chessebeuf, M., Exilie, M.F., Piard, G., Chatelain, P. and Padieu, P. (1981). Effect of serum concentration on the chromosome number of a rat liver epithelial cell lines. Cell Biol. Int. Rep.5:399–408.

    Google Scholar 

  • Chessebeuf, M., Fischbach, M. and Padieu, P. (1984). Time course study ofL-tyrosine aminotransferase induction in serum-supplemented and serum-free rat liver cell lines. Cell Biol. Toxicol.1:31–40.

    Google Scholar 

  • Chessebeuf, M. and Padieu, P. (1984). Rat liver epithelial cell cultures in a serum-free medium: primary cultures and derived cell lines expressing differentiated functions. In Vitro20:780–795.

    Google Scholar 

  • Chessebeuf-Padieu, M.:, Mignot, G., Tsaconas, C., Fischbach, M., Padieu, P. and Mack, G. (1986). Serum free and protein free medium for the continuous culture of epithelial cell lines, and myeloma and hybridoma strains. In: Therapeutic Agents Produced by Gene Engineering, pp 385–397, Medsi, Paris.

    Google Scholar 

  • Diez-Ibanez, M.A., Chessebeuf-Padieu, M., Nordmann, P. and Padieu, P. (1987). Gas chromatography-mass spectrometry analysis of N-tert-butyldime-thylsilyl derivatives of 2-acetylaminofluorene and metabolites in isolated rat hepatocytes. Cell Biol. Toxicol.3:327–330.

    Google Scholar 

  • Exilie, M.F., Turc, C., Chessebeuf, M., and Padieu, P. (1980). Nonrandom chromosome changes in nine rat liver epithelial cell lines. Cell Biology International Report4:459–469.

    Google Scholar 

  • Fischbach, M., Chessebeuf, M. and Padieu, P. (1984). Analysis of albumin and transferrin secretion of rat liver epithelial cells in culture. Biol. Cell52:47.

    Google Scholar 

  • Foliot, A., Glaise, D., Erlinger, S. and Guguen-Guillouzo, C. (1985). Long term maintenance of taurocholate uptake by adult rat hepatocytes co-cultured with a liver epithelial cell lines. Hepatology5:215–219.

    Google Scholar 

  • Franke, W.W., Mayer, D., Schmid, E., Denk, H. and Borenfreud, E. (1981). Differences of expression of cytoskeletal proteins in cultured rat hepatocytes and hepatoma cells. Exp. Cell Res.134:345–365.

    Google Scholar 

  • Grisham, J.W. (1983). Cell types in rat liver cultures: their identification and isolation. Mol. Cell. Biochem.54:23–33.

    Google Scholar 

  • Guiguet, M., Exilie-Frigere, M.F., Dethieux, M.C., Bidan, Y. and Mack, G. (1988). Biosynthesis of third component of complement in rat liver epithelial cell lines and its stimulation by effector molecules from cultured human mononuclear cells. In Vitro Cell. Develop. Biol.23:821–829.

    Google Scholar 

  • Guillouzo, A., Delers, F., Clement, B., Bernard, N. and Engler, R. (1984). Long term production of acute phase proteins by adult rat hepatocytes co-cultured with another liver cell type in serum-free medium. Biochem. Biophys. Res. Comm.120:311–317.

    Google Scholar 

  • Hietter H., Trifilieff, J.C., Richet, L., Beck, J.P., Luu, B. and Ourisson, G. (1984). Antagonistic action of cholesterol towards the toxicity of hydrosterols on cultured hepatoma cells. Biochem. Biophys. Res. Comm.120:657–664.

    Google Scholar 

  • Kandutsch, A. and Chen, H. (1973). Inhibition of sterol synthesis in cultured mouse cells by 7α-hydroxycholesterol, 7β-hydroxycholesterol, and 7-ketocholesterol. J. Biol. Chem.248:8408–8417.

    Google Scholar 

  • Mersel, M., Luu, B. and Ourisson, G. (1984). Differential action of 7β-hydroxycholesterol on myocardial cells and fibroblasts in primary cultures. Obtention of pure myocardial cells. C.R. Acad. Sci. Paris299:221–225.

    Google Scholar 

  • Padieu, P., Maume, G., Hussein, N., Chessebeuf, M. and Tsaconas, C. (1985). Biosynthesis of sterols and bile acids in rat liver epithelial cell lines. Biochem. Biophys. Acta833:245–261.

    Google Scholar 

  • Richert, L., Castagna, M., Beck, J.P., Rong, S., Luu, B. and Ourisson, G. (1984). Growth-rate related and hydroxycholesterol-induced changes in membrane fluidity of cultured hepatoma cells: correlation with 3-hydroxy-3-methylglutaryl CoA reductase activity. Biochem. Biosphys. Res. Comm.120:192–198.

    Google Scholar 

  • Riou, G. and Benard, J. (1982). The culture of human tumor cells, clinical prospects in cancer chemotherapy. Trends Pharmacol. Sci.2:78–80.

    Google Scholar 

  • Rong, S., Bergmann, C., Luu, B., Beck, J.P. and Ourisson, G. (1985). Activité antitumoralein vivo de dérivés hydrosolubles de 7-hydroxycholésterols. C.R. Acad. Sci. Paris300:89–94.

    Google Scholar 

  • Shiojiri, N. (1984). Analysis of differentiation of hepatocytes and bile duct cells in developing mouse liver by albumin immunofluorescence. Dev. Growth Differ.26:555–561.

    Google Scholar 

  • Williams, G.M. and Gunn, J.M. (1974). Long term cell culture of adult rat liver epithelial cells. Exp. Cell. Res.69:106–112.

    Google Scholar 

  • Williams, G.M., Bermudez, E. and Schramuzzino, D. (1977). Rat hepatocyte primary cell culture. III. Improved dissociation and attachment techniques and the enhancement of survival by culture medium. In Vitro13:809–817.

    Google Scholar 

  • Zander, M., Koch, P., Luu, B., Ourisson, G. and Beck, J.P. (1977). Chemistry and biochemistry of chinese drugs. III. Mechanism of hydroxylated sterols on cultured hepatoma cells. J. Chem. Res.219:2572–2584.

    Google Scholar 

Download references

Author information

Affiliations

  1. Laboratoire de Biochimie Médicale Faculté de Médecine, 7 bd Jeanne d'Arc, F-21033, Dijon

    Patrice Nordmann, Miguel Diez-Ibanez, Martine Chessebeuf-Padieu & Gerard Mack

  2. Centre de Neurochimie du CNRS et Laboratoire de Chimie Organique des Substances Naturelles associé au CNRS, 5 rue Blaise Pascal, F-67084, Strasbourg

    Bang Luu & Marcel Mersel

Authors
  1. Patrice Nordmann
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Miguel Diez-Ibanez
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Martine Chessebeuf-Padieu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Bang Luu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Gerard Mack
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Marcel Mersel
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

1. Author to whom all correspondence should be addressed.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Nordmann, P., Diez-Ibanez, M., Chessebeuf-Padieu, M. et al. Toxic effects of 70-hydroxycholesterol on rat liver primary cultures, epithelial lines and co-cultures.. Cell Biol Toxicol 5, 261–270 (1989). https://doi.org/10.1007/BF01795355

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01795355

Key words

  • 7β-hydroxycholesterol
  • cholesterol
  • hepatocytes
  • liver epithelial cells