Skip to main content
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1995 Mar;15(3):1244–1253. doi: 10.1128/mcb.15.3.1244

Repression of platelet-derived growth factor beta-receptor expression by mitogenic growth factors and transforming oncogenes in murine 3T3 fibroblasts.

C Vaziri 1, D V Faller 1
PMCID: PMC230347  PMID: 7862118

Abstract

Platelet-derived growth factor BB (PDGF-BB) is an important extracellular factor for regulating the G0-S phase transition of murine BALB/c-3T3 fibroblasts. We have investigated the expression of the PDGF beta receptor (PDGF beta R) in these cells. We show that the state of growth arrest in G0, resulting from serum deprivation, is associated with increased expression of the PDGF beta R. When the growth-arrested fibroblasts are stimulated to reenter the cell cycle by the mitogenic action of serum or certain specific combinations of growth factors, PDGF beta R mRNA levels and cell surface PDGF-BB-binding sites are markedly downregualted. Oncogene-transformed 3T3 cell lines, which fail to undergo growth arrest following prolonged serum deprivation, express constitutively low levels of the PDGF beta R mRNA and possess greatly reduced numbers of cell surface PDGF receptors, as determined by PDGF-BB binding and Western blotting (immunoblotting). Nuclear runoff assays indicate the mechanism of repression of PDGF beta R expression to be, at least in large part, transcriptional. These data indicate that expression of the PDGF beta R is regulated in a growth state-dependent manner in fibroblasts and suggest that this may provide a means by which cells can modulate their responsiveness to the actions of PDGF.

Full Text

The Full Text of this article is available as a PDF (401.5 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aharonov A., Pruss R. M., Herschman H. R. Epidermal growth factor. Relationship between receptor regulation and mitogenesis in 3T3 cells. J Biol Chem. 1978 Jun 10;253(11):3970–3977. [PubMed] [Google Scholar]
  2. Bedard P. A., Yannoni Y., Simmons D. L., Erikson R. L. Rapid repression of quiescence-specific gene expression by epidermal growth factor, insulin, and pp60v-src. Mol Cell Biol. 1989 Mar;9(3):1371–1375. doi: 10.1128/mcb.9.3.1371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Benjamin C. W., Connor J. A., Tarpley W. G., Gorman R. R. NIH-3T3 cells transformed by the EJ-ras oncogene exhibit reduced platelet-derived growth factor-mediated Ca2+ mobilization. Proc Natl Acad Sci U S A. 1988 Jun;85(12):4345–4349. doi: 10.1073/pnas.85.12.4345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Benjamin C. W., Tarpley W. G., Gorman R. R. Loss of platelet-derived growth factor-stimulated phospholipase activity in NIH-3T3 cells expressing the EJ-ras oncogene. Proc Natl Acad Sci U S A. 1987 Jan;84(2):546–550. doi: 10.1073/pnas.84.2.546. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bonin P. D., Chiou W. J., McGee J. E., Singh J. P. Two signal transduction pathways mediate interleukin-1 receptor expression in Balb/c3T3 fibroblasts. J Biol Chem. 1990 Oct 25;265(30):18643–18649. [PubMed] [Google Scholar]
  6. Bowen-Pope D. F., Ross R. Methods for studying the platelet-derived growth factor receptor. Methods Enzymol. 1985;109:69–100. doi: 10.1016/0076-6879(85)09078-4. [DOI] [PubMed] [Google Scholar]
  7. Bowen-Pope D. F., Vogel A., Ross R. Production of platelet-derived growth factor-like molecules and reduced expression of platelet-derived growth factor receptors accompany transformation by a wide spectrum of agents. Proc Natl Acad Sci U S A. 1984 Apr;81(8):2396–2400. doi: 10.1073/pnas.81.8.2396. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Buday L., Downward J. Epidermal growth factor regulates the exchange rate of guanine nucleotides on p21ras in fibroblasts. Mol Cell Biol. 1993 Mar;13(3):1903–1910. doi: 10.1128/mcb.13.3.1903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Campisi J., Pardee A. B. Post-transcriptional control of the onset of DNA synthesis by an insulin-like growth factor. Mol Cell Biol. 1984 Sep;4(9):1807–1814. doi: 10.1128/mcb.4.9.1807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cantley L. C., Auger K. R., Carpenter C., Duckworth B., Graziani A., Kapeller R., Soltoff S. Oncogenes and signal transduction. Cell. 1991 Jan 25;64(2):281–302. doi: 10.1016/0092-8674(91)90639-g. [DOI] [PubMed] [Google Scholar]
  11. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  12. Coats S. R., Olson J. E., Pledger W. J. Rapid induction of competence formation is PDGF-isoform specific. J Cell Biochem. 1992 Mar;48(3):242–247. doi: 10.1002/jcb.240480304. [DOI] [PubMed] [Google Scholar]
  13. Cook S. J., McCormick F. Inhibition by cAMP of Ras-dependent activation of Raf. Science. 1993 Nov 12;262(5136):1069–1072. doi: 10.1126/science.7694367. [DOI] [PubMed] [Google Scholar]
  14. Drozdoff V., Pledger W. J. Cellular response to platelet-derived growth factor (PDGF)-AB after down-regulation of PDGF alpha-receptors. Evidence that functional binding does not require alpha-receptors. J Biol Chem. 1991 Sep 15;266(26):17165–17172. [PubMed] [Google Scholar]
  15. Fornace A. J., Jr, Nebert D. W., Hollander M. C., Luethy J. D., Papathanasiou M., Fargnoli J., Holbrook N. J. Mammalian genes coordinately regulated by growth arrest signals and DNA-damaging agents. Mol Cell Biol. 1989 Oct;9(10):4196–4203. doi: 10.1128/mcb.9.10.4196. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gliniak B. C., Rohrschneider L. R. Expression of the M-CSF receptor is controlled posttranscriptionally by the dominant actions of GM-CSF or multi-CSF. Cell. 1990 Nov 30;63(5):1073–1083. doi: 10.1016/0092-8674(90)90510-l. [DOI] [PubMed] [Google Scholar]
  17. Gokkel E., Grossman Z., Ramot B., Yarden Y., Rechavi G., Givol D. Structural organization of the murine c-kit proto-oncogene. Oncogene. 1992 Jul;7(7):1423–1429. [PubMed] [Google Scholar]
  18. Gronwald R. G., Seifert R. A., Bowen-Pope D. F. Differential regulation of expression of two platelet-derived growth factor receptor subunits by transforming growth factor-beta. J Biol Chem. 1989 May 15;264(14):8120–8125. [PubMed] [Google Scholar]
  19. Heldin C. H. Structural and functional studies on platelet-derived growth factor. EMBO J. 1992 Dec;11(12):4251–4259. doi: 10.1002/j.1460-2075.1992.tb05523.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hicks K., Friedman B., Rosner M. R. Basic and acidic fibroblast growth factors modulate the epidermal growth factor receptor by a protein kinase C-independent pathway. Biochem Biophys Res Commun. 1989 Oct 31;164(2):796–803. doi: 10.1016/0006-291x(89)91529-5. [DOI] [PubMed] [Google Scholar]
  21. Izzo N. J., Jr, Tulenko T. N., Colucci W. S. Phorbol esters and norepinephrine destabilize alpha 1B-adrenergic receptor mRNA in vascular smooth muscle cells. J Biol Chem. 1994 Jan 21;269(3):1705–1710. [PubMed] [Google Scholar]
  22. Keating M. T., Williams L. T. Processing of the platelet-derived growth factor receptor. Biosynthetic and degradation studies using anti-receptor antibodies. J Biol Chem. 1987 Jun 5;262(16):7932–7937. [PubMed] [Google Scholar]
  23. Kitayama H., Sugimoto Y., Matsuzaki T., Ikawa Y., Noda M. A ras-related gene with transformation suppressor activity. Cell. 1989 Jan 13;56(1):77–84. doi: 10.1016/0092-8674(89)90985-9. [DOI] [PubMed] [Google Scholar]
  24. Kypta R. M., Goldberg Y., Ulug E. T., Courtneidge S. A. Association between the PDGF receptor and members of the src family of tyrosine kinases. Cell. 1990 Aug 10;62(3):481–492. doi: 10.1016/0092-8674(90)90013-5. [DOI] [PubMed] [Google Scholar]
  25. Lazarovici P., Dickens G., Kuzuya H., Guroff G. Long-term, heterologous down-regulation of the epidermal growth factor receptor in PC12 cells by nerve growth factor. J Cell Biol. 1987 Jun;104(6):1611–1621. doi: 10.1083/jcb.104.6.1611. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lee N. H., Earle-Hughes J., Fraser C. M. Agonist-mediated destabilization of m1 muscarinic acetylcholine receptor mRNA. Elements involved in mRNA stability are localized in the 3'-untranslated region. J Biol Chem. 1994 Feb 11;269(6):4291–4298. [PubMed] [Google Scholar]
  27. Leof E. B., Van Wyk J. J., O'Keefe E. J., Pledger W. J. Epidermal growth factor (EGF) is required only during the traverse of early G1 in PDGF stimulated density-arrested BALB/c-3T3 cells. Exp Cell Res. 1983 Aug;147(1):202–208. doi: 10.1016/0014-4827(83)90285-9. [DOI] [PubMed] [Google Scholar]
  28. Leof E. B., Wharton W., van Wyk J. J., Pledger W. J. Epidermal growth factor (EGF) and somatomedin C regulate G1 progression in competent BALB/c-3T3 cells. Exp Cell Res. 1982 Sep;141(1):107–115. doi: 10.1016/0014-4827(82)90073-8. [DOI] [PubMed] [Google Scholar]
  29. Lin A. H., Groppi V. E., Gorman R. R. Platelet-derived growth factor does not induce c-fos in NIH 3T3 cells expressing the EJ-ras oncogene. Mol Cell Biol. 1988 Nov;8(11):5052–5055. doi: 10.1128/mcb.8.11.5052. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Lu K., Campisi J. Ras proteins are essential and selective for the action of insulin-like growth factor 1 late in the G1 phase of the cell cycle in BALB/c murine fibroblasts. Proc Natl Acad Sci U S A. 1992 May 1;89(9):3889–3893. doi: 10.1073/pnas.89.9.3889. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Luttrell D. K., Lee A., Lansing T. J., Crosby R. M., Jung K. D., Willard D., Luther M., Rodriguez M., Berman J., Gilmer T. M. Involvement of pp60c-src with two major signaling pathways in human breast cancer. Proc Natl Acad Sci U S A. 1994 Jan 4;91(1):83–87. doi: 10.1073/pnas.91.1.83. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. McCormick F. Signal transduction. How receptors turn Ras on. Nature. 1993 May 6;363(6424):15–16. doi: 10.1038/363015a0. [DOI] [PubMed] [Google Scholar]
  33. Olwin B. B., Hauschka S. D. Identification of the fibroblast growth factor receptor of Swiss 3T3 cells and mouse skeletal muscle myoblasts. Biochemistry. 1986 Jun 17;25(12):3487–3492. doi: 10.1021/bi00360a001. [DOI] [PubMed] [Google Scholar]
  34. Pardee A. B. G1 events and regulation of cell proliferation. Science. 1989 Nov 3;246(4930):603–608. doi: 10.1126/science.2683075. [DOI] [PubMed] [Google Scholar]
  35. Parries G., Hoebel R., Racker E. Opposing effects of a ras oncogene on growth factor-stimulated phosphoinositide hydrolysis: desensitization to platelet-derived growth factor and enhanced sensitivity to bradykinin. Proc Natl Acad Sci U S A. 1987 May;84(9):2648–2652. doi: 10.1073/pnas.84.9.2648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Paulsson Y., Karlsson C., Heldin C. H., Westermark B. Density-dependent inhibitory effect of transforming growth factor-beta 1 on human fibroblasts involves the down-regulation of platelet-derived growth factor alpha-receptors. J Cell Physiol. 1993 Oct;157(1):97–103. doi: 10.1002/jcp.1041570113. [DOI] [PubMed] [Google Scholar]
  37. Pietrzkowski Z., Sell C., Lammers R., Ullrich A., Baserga R. Roles of insulinlike growth factor 1 (IGF-1) and the IGF-1 receptor in epidermal growth factor-stimulated growth of 3T3 cells. Mol Cell Biol. 1992 Sep;12(9):3883–3889. doi: 10.1128/mcb.12.9.3883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Pledger W. J., Stiles C. D., Antoniades H. N., Scher C. D. An ordered sequence of events is required before BALB/c-3T3 cells become committed to DNA synthesis. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2839–2843. doi: 10.1073/pnas.75.6.2839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Psarras S., Kletsas D., Stathakos D. Restoration of down-regulated PDGF receptors by TGF-beta in human embryonic fibroblasts. Enhanced response during cellular in vitro aging. FEBS Lett. 1994 Feb 14;339(1-2):84–88. doi: 10.1016/0014-5793(94)80390-0. [DOI] [PubMed] [Google Scholar]
  40. Quiñones M. A., Mundschau L. J., Rake J. B., Faller D. V. Dissociation of platelet-derived growth factor (PDGF) receptor autophosphorylation from other PDGF-mediated second messenger events. J Biol Chem. 1991 Jul 25;266(21):14055–14063. [PubMed] [Google Scholar]
  41. Rake J. B., Quiñones M. A., Faller D. V. Inhibition of platelet-derived growth factor-mediated signal transduction by transforming ras. Suppression of receptor autophosphorylation. J Biol Chem. 1991 Mar 15;266(8):5348–5352. [PubMed] [Google Scholar]
  42. Satoh T., Endo M., Nakafuku M., Nakamura S., Kaziro Y. Platelet-derived growth factor stimulates formation of active p21ras.GTP complex in Swiss mouse 3T3 cells. Proc Natl Acad Sci U S A. 1990 Aug;87(15):5993–5997. doi: 10.1073/pnas.87.15.5993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Scher C. D., Shepard R. C., Antoniades H. N., Stiles C. D. Platelet-derived growth factor and the regulation of the mammalian fibroblast cell cycle. Biochim Biophys Acta. 1979 Aug 10;560(2):217–241. doi: 10.1016/0304-419x(79)90020-9. [DOI] [PubMed] [Google Scholar]
  44. Scher C. D., Young S. A., Locatell K. L. Control of cytolysis of BALB/c-3T3 cells by platelet-derived growth factor: a model system for analyzing cell death. J Cell Physiol. 1982 Nov;113(2):211–218. doi: 10.1002/jcp.1041130205. [DOI] [PubMed] [Google Scholar]
  45. Schneider C., King R. M., Philipson L. Genes specifically expressed at growth arrest of mammalian cells. Cell. 1988 Sep 9;54(6):787–793. doi: 10.1016/s0092-8674(88)91065-3. [DOI] [PubMed] [Google Scholar]
  46. Shamah S. M., Stiles C. D., Guha A. Dominant-negative mutants of platelet-derived growth factor revert the transformed phenotype of human astrocytoma cells. Mol Cell Biol. 1993 Dec;13(12):7203–7212. doi: 10.1128/mcb.13.12.7203. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Stiles C. D. The molecular biology of platelet-derived growth factor. Cell. 1983 Jul;33(3):653–655. doi: 10.1016/0092-8674(83)90008-9. [DOI] [PubMed] [Google Scholar]
  48. Tamm I., Kikuchi T. Insulin-like growth factor-1 (IGF-1), insulin, and epidermal growth factor (EGF) are survival factors for density-inhibited, quiescent Balb/c-3T3 murine fibroblasts. J Cell Physiol. 1990 Jun;143(3):494–500. doi: 10.1002/jcp.1041430314. [DOI] [PubMed] [Google Scholar]
  49. Thomopoulos P., Roth J., Lovelace E., Pastan I. Insulin receptors in normal and transformed fibroblasts: relationship to growth and transformation. Cell. 1976 Jul;8(3):417–423. doi: 10.1016/0092-8674(76)90154-9. [DOI] [PubMed] [Google Scholar]
  50. Ullrich A., Schlessinger J. Signal transduction by receptors with tyrosine kinase activity. Cell. 1990 Apr 20;61(2):203–212. doi: 10.1016/0092-8674(90)90801-k. [DOI] [PubMed] [Google Scholar]
  51. Wang H., Segaloff D. L., Ascoli M. Lutropin/choriogonadotropin down-regulates its receptor by both receptor-mediated endocytosis and a cAMP-dependent reduction in receptor mRNA. J Biol Chem. 1991 Jan 15;266(2):780–785. [PubMed] [Google Scholar]
  52. Wasilenko W. J., Nori M., Testerman N., Weber M. J. Inhibition of epidermal growth factor receptor biosynthesis caused by the src oncogene product, pp60v-src. Mol Cell Biol. 1990 Mar;10(3):1254–1258. doi: 10.1128/mcb.10.3.1254. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Welham M. J., Schrader J. W. Modulation of c-kit mRNA and protein by hemopoietic growth factors. Mol Cell Biol. 1991 May;11(5):2901–2904. doi: 10.1128/mcb.11.5.2901. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Wharton W., Leof E., Pledger W. J., O'Keefe E. J. Modulation of the epidermal growth factor receptor by platelet-derived growth factor and choleragen: effects on mitogenesis. Proc Natl Acad Sci U S A. 1982 Sep;79(18):5567–5571. doi: 10.1073/pnas.79.18.5567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Williams L. T. Signal transduction by the platelet-derived growth factor receptor. Science. 1989 Mar 24;243(4898):1564–1570. doi: 10.1126/science.2538922. [DOI] [PubMed] [Google Scholar]
  56. Wyke A. W., Cushley W., Wyke J. A. Mitogenesis by v-Src: a need for active oncoprotein both in leaving G0 and in completing G1 phases of the cell cycle. Cell Growth Differ. 1993 Aug;4(8):671–678. [PubMed] [Google Scholar]
  57. Yarden Y., Escobedo J. A., Kuang W. J., Yang-Feng T. L., Daniel T. O., Tremble P. M., Chen E. Y., Ando M. E., Harkins R. N., Francke U. Structure of the receptor for platelet-derived growth factor helps define a family of closely related growth factor receptors. Nature. 1986 Sep 18;323(6085):226–232. doi: 10.1038/323226a0. [DOI] [PubMed] [Google Scholar]
  58. Yayon A., Klagsbrun M., Esko J. D., Leder P., Ornitz D. M. Cell surface, heparin-like molecules are required for binding of basic fibroblast growth factor to its high affinity receptor. Cell. 1991 Feb 22;64(4):841–848. doi: 10.1016/0092-8674(91)90512-w. [DOI] [PubMed] [Google Scholar]
  59. Yu C. L., Prochownik E. V., Imperiale M. J., Jove R. Attenuation of serum inducibility of immediate early genes by oncoproteins in tyrosine kinase signaling pathways. Mol Cell Biol. 1993 Apr;13(4):2011–2019. doi: 10.1128/mcb.13.4.2011. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Zhan X., Hu X., Friesel R., Maciag T. Long term growth factor exposure and differential tyrosine phosphorylation are required for DNA synthesis in BALB/c 3T3 cells. J Biol Chem. 1993 May 5;268(13):9611–9620. [PubMed] [Google Scholar]
  61. Zullo J. N., Faller D. V. P21 v-ras inhibits induction of c-myc and c-fos expression by platelet-derived growth factor. Mol Cell Biol. 1988 Dec;8(12):5080–5085. doi: 10.1128/mcb.8.12.5080. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES