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Regulation of surface expression of high-affinity receptors for epidermal growth factor (EGF) in hepatocytes by hormones, differentiating agents, and phorbol ester

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Abstract

Freshly isolated adult rat hepatocytes exhibit a nonhomogeneous population of epidermal growth factor (EGF) receptors with about 10,000 high-affinity binding sites (K d 20 pM) and about 200,000 low-affinity sites (K d 600 pM) per cell. With culturing as primary monolayers under conditions where the cells show a marked increase in the sensitivity to the growth-stimulatory effect of EGF, a gradual reduction in the number of EGF receptors and an almost complete loss of high-affinity EGF receptors is seen. Insulin, which promotes growth of hepatocytes in concert with EGF, enhances the down-regulation of these high-affinity receptors. The differentiating (and growth-inhibitory) agentn-butyrate counteracts this down-regulation and preserves the high-affinity receptors. This effect of butyrate is synergistic with the glucocorticoid agent dexamethasone. Another differentiating agent, dimethylsulfoxide (DMSO), also counteracts the down-regulation of high-affinity EGF receptors. Moreover, the tumor promoter, tetradecanoylphorbol acetate (TPA), down-regulates the EGF receptor. This effect is particularly evident when studying the high-affinity receptors up-regulated by prior treatment with butyrate plus dexamethyasone. Taken together these results provide strong support for the notion that an inverse relationship exists between expression of high-affinity EGF binding and responsiveness to growth activation by EGF.

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References

  1. Carpenter G: Receptors for epidermal growth factor and other polypeptide mitogens. Annu Rev Biochem 56:881–914, 1987

    Google Scholar 

  2. Schlessinger J: The epidermal growth factor receptor as a multifunctional allosteric protein. Biochemistry 27:3119–3123, 1988

    Google Scholar 

  3. Downward J, Yarden Y, Mayes E, Scrace G, Totty N, Stockwell P, Ullrich A, Schlessinger J, Waterfield MD: Close similarity of epidermal growth factor receptor andv-erb-B oncogene protein sequences. Nature 307:521–527, 1984

    Google Scholar 

  4. Thompson DM, Gill GN: The EGF receptor: Structure, regulation and potential role in malignancy. Cancer Surv 4:767–788, 1985

    Google Scholar 

  5. Pandiella A, Beguinot L, Vicentini LM, Meldolesi J: Transmembrane signalling at the epidermal growth factor receptor. Trends Pharmacol Sci 10:411–414, 1989

    Google Scholar 

  6. Davis RJ: Independent mechanisms account for the regulation by protein kinase C of the epidermal growth factor receptor affinity and tyrosine-protein kinase activity. J Biol Chem 263:9462–9469, 1988

    Google Scholar 

  7. Bellot F, Moolenaar W, Kris R, Mirakhur B, Verlaan I, Ullrich A, Schlessinger J, Felder S: High-affinity epidermal growth factor binding is specifically reduced by a monoclonal antibody, and appears necessary for early responses. J Cell Biol 110:491–502, 1990

    Google Scholar 

  8. Sand TE, Christoffersen T: Temporal requirement for epidermal growth factor and insulin in the stimulation of hepatocyte DNA synthesis. J Cell Physiol 131:141–148, 1987

    Google Scholar 

  9. Gladhaug IP, Refsnes M, Sand TE, Christoffersen T: Effects of butyrate on epidermal growth factor receptor binding, morphology, and DNA synthesis in cultured rat hepatocytes. Cancer Res 48:6560–6564, 1988

    Google Scholar 

  10. Gladhaug IP, Christoffersen T:n-Butyrate and dexamethazone synergistically modulate the surface expression of epidermal growth factor receptors in cultured rat hepatocytes. FEBS Lett 243:21–24, 1989

    Google Scholar 

  11. Gladhaug IP, Refsnes M, Christoffersen T: Regulation of hepatocyte epidermal growth factor receptors byn-butyrate and dimethyl sulfoxide: Sensitivity to modulation by the tumor promoter TPA. Anticancer Res 9:1587–1592, 1989

    Google Scholar 

  12. Seglen PO: Preparation of isolated rat liver cells. Methods Cell Biol 13:29–83, 1976

    Google Scholar 

  13. Christoffersen T, Refsnes M, Brønstad GO, Østby E, Huse J, Haffner F, Sand TE, Hunt NH, Sonne O: Changes in hormone responsiveness and cyclic AMP metabolism in rat hepatocytes during primary culture and effects of supplementing the medium with insulin and dexamethasone. Eur J Biochem 138:217–226, 1984

    Google Scholar 

  14. Sand TE, Christoffersen T: A simple medium for the study of hepatocyte growth in culture under defined conditions. In Vitro Cell Dev Biol 24:981–984, 1988

    Google Scholar 

  15. Gladhaug IP, Christoffersen T: Kinetics of epidermal growth factor binding and processing in isolated intact rat hepatocytes. Dynamic externalization of receptors during ligand internalization. Eur J Biochem 164:267–275, 1987

    Google Scholar 

  16. Sonne O, Berg T, Christoffersen T: Binding of125I-labeled glucagon and glucagon-stimulated accumulation of adenosine 3∶5-monophosphate in isolated intact rat hepatocytes. Evidence for receptor heterogeneity. J Biol Chem 253:3203–3210, 1978

    Google Scholar 

  17. Munson PJ, Rodbard D: LIGAND: A versatile computerized approach for characterization of ligand-binding systems. Anal Biochem 107:220–239, 1980

    Google Scholar 

  18. Gladhaug IP, Christoffersen T: Rapid constitutive internalization and externalization of epidermal growth factor receptors in isolated rat hepatocytes. Monensin inhibits receptor externalization and reduces the capacity for continued endocytosis of epidermal growth factor. J Biol Chem 263:12199–12203, 1988

    Google Scholar 

  19. Gladhaug IP, Christoffersen T: Amiloride inhibits constitutive internalization and increases the surface number of epidermal growth factor receptors in intact rat hepatocytes. J Cell Physiol 143:188–195, 1990

    Google Scholar 

  20. Moriarity DM, Savage R Jr: Interaction of epidermal growth factor with adult rat liver parenchymal cells in primary culture. Arch Biochem Biophys 203:506–518, 1980

    Google Scholar 

  21. Cruise JL, Cotecchia S, Michalopoulos G: Norepinephrine decreases EGF binding in primary rat hepatocyte cultures. J Cell Physiol 127:39–44, 1986

    Google Scholar 

  22. Eckl PM, Meyer SA, Whitcombe WR, Jirtle RL: Phenobarbital reduces EGF receptors and the ability of physiological concentrations of calcium to suppress hepatocyte proliferation. Carcinogenesis 9:479–483, 1988

    Google Scholar 

  23. Meyer SA, Jirtle RL: Phenobarbital decreases hepatocyte EGF receptor expression independent of protein kinase C activation. Biochem Biophys Res Commun 158:652–659, 1989

    Google Scholar 

  24. Lin Q, Blaisdell J, O'Keefe E, Earp HS: Insulin inhibits the glucocorticoid-mediated increase in hepatocyte EGF binding. J Cell Physiol 119:267–272, 1984

    Google Scholar 

  25. Kruh J: Effects of sodium butyrate, a new pharmacological agent, on cells in culture. Mol Cell Biochem 42:65–82, 1982

    Google Scholar 

  26. Engelmann GL, Staecker JL, Richardson AG: Effect of sodium butyrate on primary cultures of adult rat hepatocytes. In Vitro Cell Dev Biol 23:86–92, 1987

    Google Scholar 

  27. Staecker JL, Sattler CA, Pitot HC: Sodium butyrate preserves aspects of the differentiated phenotype of normal adult rat hepatocytes in culture. J Cell Physiol 135:367–376, 1988

    Google Scholar 

  28. McGowan JA: Reciprocal regulation of adult rat hepatocyte growth and functional activities in culture by dimethyl sulfoxide. J Cell Physiol 137:497–504, 1988

    Google Scholar 

  29. Clayton DF, Harrelson AL, Darnell JE Jr: Dependence of liver-specific transcription on tissue organization. Mol Cell Biol 5:2623–2632, 1985

    Google Scholar 

  30. Newman JD, Eckardt GS, Boyd A, Harrison LC: Induction of the insulin receptor and other differentiation markers by sodium butyrate in the Burkitt lymphoma cell, Raji. Biochem Biophys Res Commun 161:101–106, 1989

    Google Scholar 

  31. Zachary I, Rozengurt E: Modulation of the epidermal growth factor receptor by mitogenic ligands: effects of bombesin and role of protein kinase C. Cancer Surv 4:729–765, 1985

    Google Scholar 

  32. Sawada N, Staeker JL, Pitot HC: Effects of tumor-promoting agents 12-O-tetradecanoylphorbol-13-acetate and phenobarbital on DNA synthesis of rat hepatocytes in primary culture. Cancer Res 47:5665–5671, 1987

    Google Scholar 

  33. Gupta C, Hattori A, Betschart JM, Virji MA, Shinozuka H: Modulation of epidermal growth factor receptors in rat hepatocytes by two liver tumor-promoting regimens, a choline-deficient and a phenobartibal diet. Cancer Res 48:1162–1165, 1988

    Google Scholar 

  34. Harris L, Preat V, Farber E: Patterns of ligand binding to normal, regenerating, preneoplastic, and neoplastic rat hepatocytes. Cancer Res 47:3954–3958, 1987

    Google Scholar 

  35. Wollenberg GK, Harris L, Farber E, Hays MA: Inverse relationship between epidermal growth factor induced proliferation and expression of high affinity surface epidermal growth factor receptors in rat hepatocytes. Lab Invest 60:254–259, 1989

    Google Scholar 

  36. Earp HS, O'Keefe EJ: Epidermal growth factor receptor number decreases during rat liver regeneration. J Clin Invest 67:1580–1583, 1981

    Google Scholar 

  37. Francavilla A, Ove P, Polimeno L, Sciascia C, Coetzee ML, Starzl TE: Epidermal growth factor and proliferation in rat hepatocytes in primary culture isolated at different times after partial hepatectomy. Cancer Res 46:1318–1323, 1986

    Google Scholar 

  38. Shi YE, Yager JD: Effects of the liver tumor promoter ethinyl estradiol on epidermal growth factor-induced DNA synthesis and epidermal growth factor receptor levels in cultured rat hepatocytes. Cancer Res 49:3574–3580, 1989

    Google Scholar 

  39. Gregoriou M, Rees AR: Properties of a monoclonal antibody to the epidermal growth factor receptor with implications for the mechanism of action of EGF. EMBO J 3:929–937, 1984

    Google Scholar 

  40. Defize LHK, Boonstra J, Meisenhelder J, Kruijer W, Tertoolen LGJ, Tilly BC, Hunter T, Henegouwen PMP van Bergen en, Moolenaar WH, Laat SW de: Signal transduction by epidermal growth factor occurs through the subclass of high affinity receptors. J Cell Biol 109:2495–2507, 1989

    Google Scholar 

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Authors and Affiliations

  1. Department of Pharmacology, Faculty of Medicine, University of Oslo, P.O. Box 1057 Blindern, N-0316, Oslo, Norway

    Ivar P. Gladhaug, Magne Refsnes & Thoralf Christoffersen

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  1. Ivar P. Gladhaug
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  2. Magne Refsnes
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  3. Thoralf Christoffersen
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Supported by The Norwegian Cancer Society, The Nordic Insulin Fund, and The Norwegian Council for Science and the Humanities.

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Gladhaug, I.P., Refsnes, M. & Christoffersen, T. Regulation of surface expression of high-affinity receptors for epidermal growth factor (EGF) in hepatocytes by hormones, differentiating agents, and phorbol ester. Digest Dis Sci 37, 233–239 (1992). https://doi.org/10.1007/BF01308177

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  • DOI: https://doi.org/10.1007/BF01308177

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