Skip to main content
SpringerLink
Log in

Inhibitors of Protein Biosynthesis Can Stimulate Proliferation of Mouse Hepatocytes in Vitro

  • Published:
Biology Bulletin of the Russian Academy of Sciences Aims and scope Submit manuscript

Abstract

Hepatocyte proliferation in the liver regenerating after partial hepatectomy ceases when the organ is restored, and the mechanism of this phenomenon is still unclear. In the experiments on fusing hepatocytes from the regenerated mouse liver (15 days after partial hepatectomy) with NIH 3T3 mouse fibroblasts, we revealed no DNA synthesis in the nuclei of stimulated fibroblasts in heterokaryons (in the presence of hepatocyte nuclei), whereas DNA synthesis in nonfused cells was undisturbed. In this work, our purpose was to find out whether the suppression of DNA synthesis in heterokaryons could be due to the appearance in hepatocytes of some endogenous factors having an inhibitory effect on proliferation. To this end, hepatocytes from the mouse liver regenerated after partial hepatectomy were treated with cycloheximide for 1–4 h and were then fused with stimulated fibroblasts. Such a short-term treatment of hepatocytes with cycloheximide proved to result in the loss of their ability to inhibit DNA synthesis in the nuclei of stimulated or quiescent fibroblasts in heterokaryons, but hepatocytes proper actively proliferated in the medium with a low serum content (0.2%). When the mice with the liver regenerated after partial hepatectomy were treated with a single sublethal dose of cycloheximide (3 mg/kg), their hepatocytes taken two days after this treatment had no inhibitory effect. Puromycin, another inhibitor of protein synthesis, had the same effect on hepatocytes. These results may be interpreted as evidence that the final stage of liver regeneration after damage is controlled by the factors having a negative effect on cell proliferation.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

REFERENCES

  • Anzano, M.A., Roberts, A.B., and Sporn, M.B., Anchorage-Independent Growth of Primary Rat Embryo Cells Is Induced by Platelet-Derived Growth Factor and Inhibited by Type-Beta Transforming Growth Factor, J. Cell Physiol., 1986, vol. 126, pp. 312–318.

    Google Scholar 

  • Bissell, D.M., Wang, S.S., Jarnagin, W.R., and Roll, F.J., Cell-Specific Expression of Transforming Growth Factor-??in Rat Liver: Evidence for Autocrine Regulation of Hepatocyte Proliferation, J. Clin. Invest., 1995, vol. 96, pp. 447–455.

    Google Scholar 

  • Braun, L., Mead, J.E., Panzica, M., Mikumo, R., Bell, G.I., and Fausto, N., Transforming Growth Factor mRNA Increases during Liver Regeneration: A Possible Paracrine Mechanism of Growth Regulation, Proc. Natl. Acad. Sci. USA, 1988, vol. 85, pp. 1539–1543.

    Google Scholar 

  • Carr, B.I., Hayashi, I., Branum, E.L., and Moses, H.L., Inhibition of DNA Synthesis in Rat Hepatocytes by Platelet-Derived Type Beta Transforming Growth Factor, Cancer Res., 1986, vol. 46, pp. 2330–2334.

    Google Scholar 

  • Cohen, P., Holmes, C.F.B., and Tsukitani, Y., Okadaic Acid: A New Probe for the Study of Cellular Regulation, Trends Biochem. Sci., 1990, vol. 15, pp. 98–102.

    Google Scholar 

  • Danielsen, C.Ch., Mechanical Properties of Native and Reconstituted Rat Tail Tendon Collagen upon Maturation in Vitro, Mech. Ageing Dev., 1987, vol. 40, pp. 9–16.

    Google Scholar 

  • Date, M., Matsuzaki, K., Matsushita, M., Tahashi, Y., Furukawa, F., and Inoue, K., Modulation of Transforming Growth Factor Function in Hepatocytes and Hepatic Stellate Cells in Rat Liver Injury, Gut, 2000, vol. 46, pp. 719–724.

    Google Scholar 

  • Dudina-Barkovskaya, A.Ya. and Mittel'man, L.A., Producing a Primary Culture of Murine Hepatocytes, Tsitologiya, 1981, vol. 23, no.8, pp. 944–946.

    Google Scholar 

  • Earp, H.S. and O'Keefe, E.J., Epidermal Growth Factor Receptor Number Decreases during Rat Liver Regeneration, J. Clin. Invest., 1981, vol. 67, pp. 1580–1583.

    Google Scholar 

  • Edwards, D.R. and Mahadevan, L.C., Protein Synthesis Inhibitors Differentially Superinduce c-fos and c-jun by Three Distinct Mechanisms: Lack of Evidence for Labile Repressions, EMBO J., 1992, vol. 11, pp. 2415–2424.

    Google Scholar 

  • Eremeev, A.V. and Setkov, N.A., DNA Synthesis in Heterokaryons Obtained by Fusing Hepatocytes from the Regenerating Liver and NIH 3T3 fibroblasts, Dokl. Ross. Akad. Nauk, 2000, vol. 372, pp. 836–839.

    Google Scholar 

  • Fabrikant, J.I., Size of Proliferating Pools in Regenerating Liver, Exp. Cell Res., 1969a, vol. 55, pp. 277–279.

    Google Scholar 

  • Fabrikant, J.I., Studies on Cell Population Kinetics in the Regenerating Liver, Natl. Cancer Inst. Monogr., 1969b, vol.?30, pp. 169–183.

    Google Scholar 

  • Fausto, N., Laird, A.D., and Webber, E.M., Role of Growth Factors and Cytokines in Hepatic Regeneration, FASEB J., 1995, vol. 9, pp. 1527–1536.

    Google Scholar 

  • Filipova, G.N., Snitkovskii, A.V., Boikov, P.Ya., and Alekseenko, A.V., Oncogene Expression in the Rat Liver upon Template Syntheses Uncoupling by Sublethal Doses of Cycloheximide, Mol. Biol. (Moscow), 1989, vol. 23, no.3, pp. 843–849.

    Google Scholar 

  • Gressner, A.M. and Wulbrand, U., Variation of Immunocytochemical Expression of Transforming Growth Factor (TGF)-??in Hepatocytes in Culture and Liver Slices, Cell Tissue Res., 1996, vol. 287, pp. 143–152.

    Google Scholar 

  • Gressner, G., Zoremba, M., and Gressner, A.M., Transforming Growth Factor Beta (TGF-?) Expression in Isolated and Cultured Rat Hepatocytes, J. Cell Physiol., 1996, vol. 167, pp. 394–405.

    Google Scholar 

  • Higgins, G. and Anderson, R., Experimental Pathology of the Liver: Restoration of the Liver of the White Rat Following Partial Surgical Removal, Arch. Pathol., 1931, vol. 12, pp.?186–202.

    Google Scholar 

  • Hugget, A.C., Krutzsch, N.C., and Thorgeirsson, S.S., Characterization of a Hepatic Proliferation Inhibitor (HPI): Effect of HPI on the Growth of Normal Liver Cells- Comparison with Transforming Growth Factor Beta, J. Cell Biochem., 1987, vol. 35, pp. 305–314.

    Google Scholar 

  • Iype, P.T. and McMahon, J.B., Hepatic Proliferation Inhibitor, Mol. Cell. Biochem., 1984, vol. 59, pp. 57–80.

    Google Scholar 

  • Lindroos, P.M., Zarnegar, R., and Michalopoulos, G.K., Hepatocyte Growth Factor (Hepatopoietin A) Rapidly Increases in Plasma before DNA Synthesis and Liver Regeneration Stimulated by Partial Hepatectomy and Carbon Tetrachloride Administration, Hepatology (St. Louis), 1991, vol. 13, pp. 743–750.

    Google Scholar 

  • Mahadevan, L.C. and Edwards, D.R., Signalling and Superinduction, Nature (London), 1991, vol. 349, pp. 747–748.

    Google Scholar 

  • Manteifel', V.M. and Zelenin, A.V., Ultrastructural Rearrangements in Guinea Pig Hepatocytes 2.5 Hours after Partial Hepatectomy, Tsitologiya, 1977, vol. 18, no.9, pp. 1063–1067.

    Google Scholar 

  • Masson, S., Scotte, M., Francois, A., Coeffier, M., Provot, F., Hiron, M., Teniere, P., Fallu, J., Salier, J.P., and Daveau, M., Changes in Growth Factor and Cytokine mRNA Levels after Hepatectomy in Rat with CCI4-Induced Cirrhosis, Am. J.?Physiol., 1999, vol. 277, pp. 838–846.

    Google Scholar 

  • McMahon, J.B., Farrelly, J.G., and Iype, P.T., Purification and Properties of a Rat Liver Protein That Specifically Inhibits the Proliferation of Nonmalignant Epithelial Cells From Rat Liver, Proc. Natl. Acad. Sci. USA, 1982, vol. 79, pp. 456–460.

    Google Scholar 

  • McMahon, J.B., Rechards, W.L., Del Campo, A.A., Song, M.-K., and Thorgeirsson, S.S., Differential Effects of Transforming Growth Factor-Beta on Proliferation of Normal and Malignant Rat Liver Epithelial Cells in Culture, Cancer Res., 1986, vol. 46, pp. 4665–4671.

    Google Scholar 

  • Michalopoulos, G.K., Liver Regeneration: Molecular Mechanisms of Growth Control, FASEB J., 1990, vol. 4, pp. 176–187.

    Google Scholar 

  • Michalopoulos, G.K. and DeFrances, M.C., Liver Regeneration, Science, 1997, vol. 276, pp. 60–66.

    Google Scholar 

  • Nakamura, T., Tomita, Y., and Ishihara, A., Density Dependent Growth Control of Adult Rat Hepatocytes in Primary Culture, J. Biochem. (Tokyo), 1983a, vol. 94, pp. 1029–1036.

    Google Scholar 

  • Nakamura, T., Yoshimoto, K., Nakayama, Y., Tomita, Y., and Ishihara, A., Reciprocal Modulation of Growth and Differentiated Functions of Mature Rat Hepatocytes in Primary Culture by Cell-Cell Contact and Cell Membranes, Proc. Natl. Acad. Sci. USA, 1983b, vol. 80, pp. 7229–7233.

    Google Scholar 

  • Nakamura, T., Tomita, Y., Hirai, R., Yamaoka, K., Kaji, K., and Ichihara, A., Inhibitory Effect of Transforming Growth Factor-??on DNA Synthesis of Adult Rat Hepatocytes in Primary Culture, Biochem. Biophys. Res. Commun., 1985, vol. 133, pp.1042–1050.

    Google Scholar 

  • Rosenwald, I.B., Setkov, N.A., Kazakov, V.N., Chen, J.-J., Ryazanov, A.G., London, I.M., and Epifanova, O.I., Transient Inhibition of Protein Synthesis Induces Expression of Proto-Oncogenes and Stimulates Resting Cells to Enter the Cell Cycle, Cell Prolif., 1995, vol. 28, pp. 631–644.

    Google Scholar 

  • Rubin, R.A., O'Keefe, E.J., and Earp, H.S., Alteration of Epidermal Growth Factor-Dependent Phosphorylation during Rat Liver Regeneration, Proc. Natl. Acad. Sci. USA, 1982, vol. 79, pp. 776–780.

    Google Scholar 

  • Russell, W.E., Coffey, R.J., Quelletta, A.J., and Moses, H.L., Type ??Transforming Growth Factor Reversibly Inhibits the Early Proliferation Response to Partial Hepatectomy in the Rat, Proc. Natl. Acad. Sci. USA, 1988, vol. 85, pp. 5126–5132.

    Google Scholar 

  • Setkov, N.A. and Eremeev, A.V., Hepatocytes in Heterokaryons Can Inhibit the Entry of Murine NIH 3T3 Fibroblast Nuclei into the S Phase, Tsitologiya, 2001, vol. 43, no.6, pp.?33–40.

    Google Scholar 

  • Setkov, N.A., Polunovskii, V.A., and Epifanova, O.I., DNA Synthesis in the Nuclei of Proliferating and Quiescent NIH 3T3 Cells in Monokaryons, Homokaryons, and Heterokaryons, Tsitologiya, 1984, vol. 26, no.6, pp. 681–689.

    Google Scholar 

  • Setkov, N.A., Kazakov, V.N., and Andreeva, T.V., Hepatocytes in Heterokaryons Do Not Retard the Entry of Stimulated Fibroblast Nuclei into the S Phase, Tsitologiya, 1991, vol. 38, no.6, pp. 76–85.

    Google Scholar 

  • Setkov, N.A., Kazakov, V.N., Rozenwald, I.B., Makarova, G.F., and Epifanova, O.I., Protein Synthesis Inhibitors, Like Growth Factors, May Render Resting 3T3 Cells Competent for DNA Synthesis: A Radioautographic Study, Cell Prolif., 1992, vol. 25, pp. 181–192.

    Google Scholar 

  • Strain, A.J., Frazer, A., Hill, D.J., and Miller, R.D., Transforming Growth Factor Beta Inhibits DNA Synthesis in Hepatocytes Isolated from Normal and Regenerating Rat Liver, Biochem. Biophys. Res. Commun., 1987, vol. 145, pp.?436–442.

    Google Scholar 

  • S'yakste, N.I., Specific Features of the Molecular Organization of Cell Nucleus Components at Different Phases of the Mitotic Phases and in Quiescent Cells, Tsitologiya, 1992, vol. 34, no.9, pp. 3–16.

    Google Scholar 

  • Thompson, N.L., Mead, J.E., Braun, L., Goeytte, M., Shank, P.R., and Fausto, N., Sequential Protooncogene Expression during Rat Liver Regeneration, Cancer. Res., 1986, vol. 46, pp. 3111–3117.

    Google Scholar 

  • Todorov, I.N., Boikov, P.Ya., Sidorenko, L.I., Chirkov, G.P., and Terskikh, V.V., Stimulation of DNA Replication in Rat Hepatocytes as a Results of Protein Synthesis Inhibition, Dokl. Akad. Nauk SSSR, 1978, vol. 239, pp. 1255–1258.

    Google Scholar 

  • Wollenberg, G.K., Stemple, E., Quinn, B.A., and Hayes, A.M., Inhibition of Proliferation of Normal, Preneoplastic and Neoplastic Rat Hepatocytes by Transforming Growth Factor Beta, Cancer. Res., 1987, vol. 47, pp. 6595–6601.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Russian Academy of Sciences, Institute of Biophysics, Siberian Branch, Akademgorodok, Krasnoyarsk, 660036, Russia

    N. A. Setkov & A. V. Eremeev

Authors
  1. N. A. Setkov
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A. V. Eremeev
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Setkov, N.A., Eremeev, A.V. Inhibitors of Protein Biosynthesis Can Stimulate Proliferation of Mouse Hepatocytes in Vitro . Biology Bulletin 30, 212–219 (2003). https://doi.org/10.1023/A:1023843409416

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1023843409416

Keywords

Search

Navigation