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Expression of c-fos and c-myc protooncogenes in an immortalized hepatocyte line harbouring SV40 T antigen and hGH as transgenes

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Abstract

A clonal hepatocyte line (FMH-202-2), derived from livers of fetal transgenic mice harbouring human growth hormone (hGH) and SV40 T antigen as transgenes, was used in the investigation of protooncogene expression involved in liver-specific growth control and/or in hepatocellular transformation. In this model system, representing an immortalized, yet untransformed phenotype, the transgenes hGH and SV40 T antigen were expressed constitutively. The c-fos protooncogene was induced by incubation with insulin, epidermal growth factor (EGF) and insulin-like growth factor (IGF-I) in a transient manner comparable to its expression in primary murine hepatocytes. Elucidation of second messenger mechanisms demonstrated that c-fos induction by hepatotrophic growth factors was not mediated by protein kinase C. In contrast to primary hepatocytes, the c-myc protooncogene exhibited a constitutive expression pattern which was independent of growth factor stimulation. These results indicate that apart from hGH and SV40 T antigen, c-myc may play a role in cellular immortalization, but that constitutive expression of these genes, even in combined coexpression, does not suffice to induce the transformed phenotype.

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References

  • Andrews, G.K., Harding, M.A., Calvet, J.P. and Adamson, E.D. (1987) The heat shock response in HeLa cells is accompanied by elevated expression of the c-fos proto-oncogene.Mol. Cell. Biol. 7, 3452–58.

    PubMed  Google Scholar 

  • Angel, P., Rahmsdorf, H.J., Pöting, A. and Herrlich, P. (1985) C-fos mRNA levels in primary human fibroblasts after arrest in various stages of the cell cycle.Cancer Cells 3, 315–9.

    Google Scholar 

  • Berczi, I., Nagy, E., De Toledo, S.M., Matusik, R.J. and Friesen, H.G. (1991) Pituitary hormones regulate c-myc and DNA synthesis in lymphoid tissue.J. Immunol. 146, 2201–6.

    PubMed  Google Scholar 

  • Blackshear, P. (1989) Insulin-stimulated protein biosynthesis as a paradigm of protein kinase C-independent growth factor action.Clin. Res. 37, 15–25.

    PubMed  Google Scholar 

  • Blackwood, E.M. and Eisenman, R.N. (1991) Max: a helix-loophelix zipper protein that forms a sequence-specific DNA binding complex with MYC.Science 251, 1211–17.

    PubMed  Google Scholar 

  • Bravo, R. and Miller, R. (1986) Involvement of proto-oncogenes in growth control: the induction of c-fos and c-myc by growth factors. In Kahn, P. and Graf, T. eds,Oncogenes and Growth Control, pp. 253–8. Berlin/Heidelberg: Springer-Verlag.

    Google Scholar 

  • Bravo, R., Neuberg, M., Burckhardt, J., Almendral, J., Wallich, R. and Müller, R. (1987) Involvement of common and cell type-specific pathways of c-fos gene control: stable induction by cAMP in macrophages.Cell 48, 251–60.

    PubMed  Google Scholar 

  • Büscher, M., Rahmsdorf, H.J., Litfin, M., Karin, M. and Herrlich, P. (1988) Activation of the c-fos gene by UV and phorbol ester: different signal transduction pathways converge to the same enhancer element.Oncogene 3, 301–11.

    PubMed  Google Scholar 

  • Chirgwin, J.M., Przybyla, A.F., MacDonald, R.J., and Rutter, W.J. (1979) Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease.Biochemistry 18, 5294–9.

    PubMed  Google Scholar 

  • Clayton, D.F. and Darnell, J.E., Jr (1983) Changes in liver-specific compared to common gene transcription during primary culture of mouse hepatocytes.Mol. Cell. Biol. 3, 1552–61.

    PubMed  Google Scholar 

  • Colby, W.W. and Shenk, T. (1982) Fragments of the simian virus 40 transforming gene facilitate transformation of rat embryo cells.Proc. Natl. Acad. Sci. USA 79, 5189–93.

    PubMed  Google Scholar 

  • Curran, T. and Franza, R., Jr (1988)FOS andJUN: the AP-1 connection.Cell 55, 395–7.

    PubMed  Google Scholar 

  • Dani, C., Blanchard, J.M., Piechaczyk, M., El Sabouty, S., Marty, L. and Jeanteur, P. (1984) Extreme instability of themyc mRNA in normal and transformed human cells.Proc. Natl. Acad. Sci. USA 81, 7046–50.

    PubMed  Google Scholar 

  • Dyer, K.R. and Messing, A. (1989) Metal-inducible pathology in the liver, pancreas, and kidney of transgenic mice expressing SV40 early region genes.Am. J. Pathol. 135, 401–10.

    PubMed  Google Scholar 

  • Fausto, N. and Mead, J. (1989) Biology of disease: regulation of liver growth: protooncogenes and transforming growth factors.Lab. Invest. 60, 4–13.

    PubMed  Google Scholar 

  • Feinberg, A.P. and Vogelstein, B. (1983) A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity.Analyt. Biochem. 132, 6–13.

    PubMed  Google Scholar 

  • Feinberg, A.P. and Vogelstein, B. (1984) Addendum: ‘A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity.Analyt. Biochem. 137, 266–7.

    PubMed  Google Scholar 

  • Fisch, T.M., Prywes, R. and Roeder, R.G. (1987) C-fos sequences necessary for basal expression and induction by epidermal growth factor, 12-0-tetradecanoyl phorbol-13-acetate, and the calcium ionophore.Mol. Cell. Biol. 7, 3490–502.

    PubMed  Google Scholar 

  • Grausz, J.D., Fradelizi, D., Dautry, F., Monier, R. and Lehn, P. (1986) Modulation of c-fos and c-myc mRNA levels in normal human lymphocytes by calcium ionophore A23187 and phorbol ester.Eur. J. Immunol. 16, 1217–21.

    PubMed  Google Scholar 

  • Greenberg, M.E. and Ziff, E.B. (1984) Stimulation of 3T3 cells induces transcription of the c-fos proto-oncogene.Nature 311, 433–8.

    PubMed  Google Scholar 

  • Hayashi, K., Makino, R. and Sugimura, T. (1984) Amplification and overexpression of the c-myc gene in Morris hepatomas.Gann. 75, 475–8.

    PubMed  Google Scholar 

  • Hoffmann, B., Piasecki, A. and Paul, D. (1989) Proliferation of fetal rat hepatocytes in response to growth factors and hormones in primary cultureJ. Cell. Physiol. 139, 654–62.

    PubMed  Google Scholar 

  • Höhne, M., Becker-Rabbenstein, V., Kahl, G.F. and Taniguchi, H. (1990a) Regulation of cytochrome P-450 CYP IA1 gene expression and protooncogene expression by growth factors in primary hepatocytes.FEBS Lett. 273, 219–22.

    PubMed  Google Scholar 

  • Höhne, M., Schaefer, S., Seifer, M., Feitelson, M.A., Paul, D. and Gerlich, W.H. (1990b) Malignant transformation of immortalized transgenic hepatocytes after transfection with hepatitis B virus DNA.EMBO J. 9, 1137–45.

    PubMed  Google Scholar 

  • Hsieh, L.L., Hsiao, W.-L., Peraino, C., Maronpot, R.R. and Weinstein, I.B. (1987) Expression of retroviral sequences and oncogenes in rat liver tumors induced by diethylnitrosamine.Cancer Res. 47, 3421–4.

    PubMed  Google Scholar 

  • Huber, B.E. and Thorgeirsson, S. (1987) Analysis of c-myc expression in a human hepatoma cell line.Cancer Res. 47, 3414–20.

    PubMed  Google Scholar 

  • Klein, G. (1986) Multible factors involved in B-cell tumorigenesis. In Kahn, P. and Graf, T. eds,Oncogenes and Growth Control, pp. 320–5. Berlin/Heidelberg: Springer-Verlag.

    Google Scholar 

  • Kouzarides, T., and Ziff, E. (1989) Behind theFOS andJUN leucine zipper.Cancer Cells 1, 71–6.

    PubMed  Google Scholar 

  • Kruijer, W., Skelly, H., Botteri, F., van der Putten, H., Barber, J.R., Verma, I.M. and Leffert, H.L. (1986) Proto-oncogene expression in regenerating liver is stimulated in cultures of primary adult rat hepatocytes.J. Biol. Chem. 261, 7929–33.

    PubMed  Google Scholar 

  • Land, H., Chen, A.C., Morgenstern, J.P., Parada, L.F. and Weinberg, R.A. (1986) Behavior ofmyc andras oncogenes in transformation of rat embryo fibroblasts.Moll Cell. Biol. 6, 1917–25.

    Google Scholar 

  • Lautenberger, J.A., Schulz, R.A., Garon, C.F., Tsichlis, P.N. and Papas, T.S. (1981) Molecular cloning of avian myelocytomatosis virus (MC29) transforming sequences.Proc. Natl Acad. Sci. USA 78, 15`8–22.

    Google Scholar 

  • Little, C.D., Nan, M.M., Carney, D.N., Gazdar, A.F. and Minna, J.D. (1983) Amplification and expression of the c-myc oncogene in human lung cancer cell lines.Nature 306, 194–6.

    PubMed  Google Scholar 

  • Mathews, L.S., Norstedt, G. and Palmiter, R.D. (1986) Regulation of insulin-like growth factor I gene expression by growth hormone.Proc. Natl Acad. Sci. USA 83, 9343–7.

    PubMed  Google Scholar 

  • Messing, A., Chen, H.Y., Palmiter, R.D. and Brinster, R.L. (1985) Peripheral neuropathies, hepatocellular carcinomas and islet cell adenomas in transgenic mice.Nature 316, 461–3.

    PubMed  Google Scholar 

  • Miller, H., Asselin, C., Dufort, D., Yang, J.-Q., Gupta, K., Marcu, K.B. and Nepveu, A. (1989) Acis acting element in the promoter region of the murine c-myc gene is necessary for transcriptional block.Mol. Cell. Biol. 9, 5340–9.

    PubMed  Google Scholar 

  • Müller, R., Bravo, R., Burckhardt, J. and Curran, T. (1984) Induction of c-fos gene and protein by growth factors precedes activation of c-myc.Nature 312, 716–21.

    PubMed  Google Scholar 

  • Norstedt, G., Levinovitz, A., Möller, C., Eriksson, L.C. and Andersson, G. (1988) Expression of insulin-like growth factor I (IGF-I) and IFG-II mRNA during hepatic development, proliferation and carcinogenesis in the rat.Carcinogenesis 9, 209–13.

    PubMed  Google Scholar 

  • Orian, J.M., Tamakoshi, K., Mackay, I.R. and Brandon, M.R. (1990) New murine model for hepatocellular carcinoma: Transgenic mice expressing metallothionein-ovine growth hormone fusion gene.J. Natl Cancer Inst. 82, 393–8.

    PubMed  Google Scholar 

  • Palmiter, R.D., Chen, H.Y., Messing, A. and Brinster, R.L. (1985) SV40 enhancer and large-T antigen are instrumental in development of choroid plexus tumors in transgenic mice.Nature 316, 457–60.

    PubMed  Google Scholar 

  • Paul, D., Höhne, M., Pinkert, C., Piasecki, A., Ummelmann, E. and Brinster, R.L. (1988) Immortalized differentiated hepatocyte lines derived from transgenic mice harboring SV40 T-antigen genes.Exp. Cell Res. 175, 354–62.

    PubMed  Google Scholar 

  • Piechaczyk, M., Yang, J., Blanchard, J., Jeanteur, P. and Marcu, K.B. (1985) Posttranscriptional mechanisms are responsible for accumulation of truncated c-myc mRNAs in murine plasma cell tumors.Cell 42, 589–97.

    PubMed  Google Scholar 

  • Porsch Hällström, I., Gustafsson, J.-A. and Blanck, A. (1990) Hypothalamo-pituitary regulation of the c-myc gene in rat liver.J. Mol. Endocrinol. 5, 267–74.

    PubMed  Google Scholar 

  • Ran, W., Dean, M., Levine, R.A., Henkle, C. and Campisi, J. (1986) Induction of c-fos and c-myc mRNA by epidermal growth factor or calcium ionophore is cAMP dependent.Proc. Natl Acad. Sci. USA 83, 8216–20.

    PubMed  Google Scholar 

  • Sandgren, E.P., Quaife, C.J., Pinkert, C.A., Palmiter, R.D. and Brinster, R.L. (1989) Oncogene-induced liver neoplasia in transgenic mice.Oncogene 4, 715–24.

    PubMed  Google Scholar 

  • Sepulveda, A.R., Finegold, M.J., Smith, B., Slagle, B.L., DeMayo, J.L., Shen, R.-F., Woo, S.L.C. and Butei, J.S. (1989) Carcinogenesis using tissue-specific expression of SV40 large T-antigen controlled by regulatory elements of the human α-1-antitrypsin gene.Cancer Res. 49, 6108–17.

    PubMed  Google Scholar 

  • Shibanuma, M., Kuroki, T. and Nose, K. (1987) Inhibition of proto-oncogene c-fos transcription by inhibitors of protein kinase C and ion transport.Eur. J. Biochem. 164, 15–9.

    PubMed  Google Scholar 

  • Southern, E.M. (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis.J. Mol. Biol. 98, 503–17.

    PubMed  Google Scholar 

  • Southern, P.J. and Berg, D. (1982) Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter.J. Mol. Genet. 1, 327–41.

    Google Scholar 

  • Spencer, C.A. and Groudine, M. (1991) Control of c-myc regulation in normal and neoplastic cells.Adv. Cancer Res. 56, 1–48.

    PubMed  Google Scholar 

  • Stumpo, D.J. and Blackshear, P.J. (1986) Insulin and growth factor effects on c-fos expression in normal and protein kinase C-deficient 3T3-L1 fibroblasts and adipocytes.Proc. Natl Acad. Sci. USA 83, 9453–7.

    PubMed  Google Scholar 

  • Taylor, S.I. (1988) Receptors for insulin and insulin-like growth factors. In Arias, I.M., Jacoby, W.B., Popper, H., Schachter, D. and Shafritz, D. eds,The Liver: Biology and Pathobiology, 2nd edition, pp. 753–67. New York: Raven Press Ltd.

    Google Scholar 

  • Thompson, N.L., Mead, J.E., Braun, L., Goyette, M., Shank, P.R. and Fausto, N. (1986) Sequential protooncogene expression during rat liver regeneration.Cancer Res. 46, 3111–7.

    PubMed  Google Scholar 

  • Van Straaten, F., Muller, R., Curran, T., Van Beveren, C. and Verma, I.M. (1983) Complete nucleotide sequence of a human c-onc gene: deduced amino acid sequence of the human c-fos protein.Proc. Natl Acad. Sci. USA 80, 3183–7.

    PubMed  Google Scholar 

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Authors and Affiliations

  1. Institute of Pharmacology and Toxicology, University of Göttigen, Göttingen, Germany

    Karen I. Hirsch-Ernst, Georg Friedrich Kahl & Martin W. Höhne

  2. Department of Cell Biology, Fraunhofer-Institut for Toxicology, Hannover, Germany

    Dieter Paul

  3. Institute of Medical Virology, University of Giessen Medical School, Frankfurter Strasse 107, D-6300, Giessen, Germany

    Martin W. Höhne

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  1. Karen I. Hirsch-Ernst
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  2. Dieter Paul
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  4. Martin W. Höhne
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Hirsch-Ernst, K.I., Paul, D., Kahl, G.F. et al. Expression of c-fos and c-myc protooncogenes in an immortalized hepatocyte line harbouring SV40 T antigen and hGH as transgenes. Transgenic Research 2, 101–108 (1993). https://doi.org/10.1007/BF01969383

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  • DOI: https://doi.org/10.1007/BF01969383

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