Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1996 Jan 1;132(1):239–249. doi: 10.1083/jcb.132.1.239

Extracellular matrix alters PDGF regulation of fibroblast integrins

PMCID: PMC2120701  PMID: 8567727

Abstract

Extracellular matrix (ECM) and growth factors are potent regulators of cell phenotype. These biological mediators of cellular responses are potentially interactive and as such could drive cells through progressive phenotypes to create new tissue as in morphogenesis and wound repair. In fact, ECM composition changes during tissue formation accompanied by alterations in cell growth and migration. How alterations in the ECM regulate cell activities is poorly defined. To address this question in wound repair, we cultured normal human dermal skin fibroblasts in relaxed collagen gels, fibronectin-rich cultures or stressed fibrin gels, and stressed collagen gels to model normal dermis, early wound provisional matrix, and late granulation tissue, respectively. Integrin subunits, alpha 2, alpha 3, and alpha 5, that define receptor specificity for collagen and provisional matrix, respectively, were measured at mRNA steady-state level before and after stimulation with platelet-derived growth factor-BB (PDGF-BB), a potent mitogen and chemoattractant for fibroblasts. Fibronectin-rich cultures and fibrin gels supported PDGF-BB induction of alpha 3 and alpha 5 mRNA. In contrast, both stressed and relaxed collagen attenuated these responses while promoting maximal alpha 2 mRNA expression. Posttranscriptional regulation was an important mechanism in this differential response. Together PDGF-BB and collagen gels promoted alpha 2, but not alpha 3 and alpha 5, mRNA stability. Conversely, when fibroblasts were in fibronectin-rich cultures, PDGF-BB promoted alpha 3 and alpha 5, but not alpha 2, mRNA stability. We suggest that ECM alterations during wound healing or any new tissue formation cause cells to respond differently to repeated growth factor stimuli. An ordered progression of cell phenotypes results, ultimately consummating tissue repair or morphogenesis.

Full Text

The Full Text of this article is available as a PDF (2.2 MB).

Selected References

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

  1. Ahlén K., Rubin K. Platelet-derived growth factor-BB stimulates synthesis of the integrin alpha 2-subunit in human diploid fibroblasts. Exp Cell Res. 1994 Dec;215(2):347–353. doi: 10.1006/excr.1994.1351. [DOI] [PubMed] [Google Scholar]
  2. Ansel J. C., Tiesman J. P., Olerud J. E., Krueger J. G., Krane J. F., Tara D. C., Shipley G. D., Gilbertson D., Usui M. L., Hart C. E. Human keratinocytes are a major source of cutaneous platelet-derived growth factor. J Clin Invest. 1993 Aug;92(2):671–678. doi: 10.1172/JCI116636. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Argraves W. S., Suzuki S., Arai H., Thompson K., Pierschbacher M. D., Ruoslahti E. Amino acid sequence of the human fibronectin receptor. J Cell Biol. 1987 Sep;105(3):1183–1190. doi: 10.1083/jcb.105.3.1183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Aumailley M., Mann K., von der Mark H., Timpl R. Cell attachment properties of collagen type VI and Arg-Gly-Asp dependent binding to its alpha 2(VI) and alpha 3(VI) chains. Exp Cell Res. 1989 Apr;181(2):463–474. doi: 10.1016/0014-4827(89)90103-1. [DOI] [PubMed] [Google Scholar]
  5. Bellas R. E., Bendori R., Farmer S. R. Epidermal growth factor activation of vinculin and beta 1-integrin gene transcription in quiescent Swiss 3T3 cells. Regulation through a protein kinase C-independent pathway. J Biol Chem. 1991 Jun 25;266(18):12008–12014. [PubMed] [Google Scholar]
  6. Berthod F., Hayek D., Damour O., Collombel C. Collagen synthesis by fibroblasts cultured within a collagen sponge. Biomaterials. 1993 Aug;14(10):749–754. doi: 10.1016/0142-9612(93)90039-5. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Clark R. A., Folkvord J. M., Hart C. E., Murray M. J., McPherson J. M. Platelet isoforms of platelet-derived growth factor stimulate fibroblasts to contract collagen matrices. J Clin Invest. 1989 Sep;84(3):1036–1040. doi: 10.1172/JCI114227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Clark R. A., Nielsen L. D., Welch M. P., McPherson J. M. Collagen matrices attenuate the collagen-synthetic response of cultured fibroblasts to TGF-beta. J Cell Sci. 1995 Mar;108(Pt 3):1251–1261. doi: 10.1242/jcs.108.3.1251. [DOI] [PubMed] [Google Scholar]
  10. Clark R. A. Regulation of fibroplasia in cutaneous wound repair. Am J Med Sci. 1993 Jul;306(1):42–48. doi: 10.1097/00000441-199307000-00011. [DOI] [PubMed] [Google Scholar]
  11. Deuel T. F., Kawahara R. S., Mustoe T. A., Pierce A. F. Growth factors and wound healing: platelet-derived growth factor as a model cytokine. Annu Rev Med. 1991;42:567–584. doi: 10.1146/annurev.me.42.020191.003031. [DOI] [PubMed] [Google Scholar]
  12. Eckes B., Mauch C., Hüppe G., Krieg T. Downregulation of collagen synthesis in fibroblasts within three-dimensional collagen lattices involves transcriptional and posttranscriptional mechanisms. FEBS Lett. 1993 Mar 1;318(2):129–133. doi: 10.1016/0014-5793(93)80006-g. [DOI] [PubMed] [Google Scholar]
  13. Elices M. J., Urry L. A., Hemler M. E. Receptor functions for the integrin VLA-3: fibronectin, collagen, and laminin binding are differentially influenced by Arg-Gly-Asp peptide and by divalent cations. J Cell Biol. 1991 Jan;112(1):169–181. doi: 10.1083/jcb.112.1.169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Enenstein J., Waleh N. S., Kramer R. H. Basic FGF and TGF-beta differentially modulate integrin expression of human microvascular endothelial cells. Exp Cell Res. 1992 Dec;203(2):499–503. doi: 10.1016/0014-4827(92)90028-7. [DOI] [PubMed] [Google Scholar]
  15. Folkman J., Moscona A. Role of cell shape in growth control. Nature. 1978 Jun 1;273(5661):345–349. doi: 10.1038/273345a0. [DOI] [PubMed] [Google Scholar]
  16. Folkvord J. M., Viders D., Coleman-Smith A., Clark R. A. Optimization of immunohistochemical techniques to detect extracellular matrix proteins in fixed skin specimens. J Histochem Cytochem. 1989 Jan;37(1):105–113. doi: 10.1177/37.1.2461979. [DOI] [PubMed] [Google Scholar]
  17. Gailit J., Clark R. A. Wound repair in the context of extracellular matrix. Curr Opin Cell Biol. 1994 Oct;6(5):717–725. doi: 10.1016/0955-0674(94)90099-x. [DOI] [PubMed] [Google Scholar]
  18. Grinnell F. Fibroblasts, myofibroblasts, and wound contraction. J Cell Biol. 1994 Feb;124(4):401–404. doi: 10.1083/jcb.124.4.401. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Grinnell F., Fukamizu H., Pawelek P., Nakagawa S. Collagen processing, crosslinking, and fibril bundle assembly in matrix produced by fibroblasts in long-term cultures supplemented with ascorbic acid. Exp Cell Res. 1989 Apr;181(2):483–491. doi: 10.1016/0014-4827(89)90105-5. [DOI] [PubMed] [Google Scholar]
  20. Gullberg D., Gehlsen K. R., Turner D. C., Ahlén K., Zijenah L. S., Barnes M. J., Rubin K. Analysis of alpha 1 beta 1, alpha 2 beta 1 and alpha 3 beta 1 integrins in cell--collagen interactions: identification of conformation dependent alpha 1 beta 1 binding sites in collagen type I. EMBO J. 1992 Nov;11(11):3865–3873. doi: 10.1002/j.1460-2075.1992.tb05479.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Haas T. A., Plow E. F. Integrin-ligand interactions: a year in review. Curr Opin Cell Biol. 1994 Oct;6(5):656–662. doi: 10.1016/0955-0674(94)90091-4. [DOI] [PubMed] [Google Scholar]
  22. Hart C. E., Forstrom J. W., Kelly J. D., Seifert R. A., Smith R. A., Ross R., Murray M. J., Bowen-Pope D. F. Two classes of PDGF receptor recognize different isoforms of PDGF. Science. 1988 Jun 10;240(4858):1529–1531. doi: 10.1126/science.2836952. [DOI] [PubMed] [Google Scholar]
  23. Heino J., Ignotz R. A., Hemler M. E., Crouse C., Massagué J. Regulation of cell adhesion receptors by transforming growth factor-beta. Concomitant regulation of integrins that share a common beta 1 subunit. J Biol Chem. 1989 Jan 5;264(1):380–388. [PubMed] [Google Scholar]
  24. Heino J., Massagué J. Transforming growth factor-beta switches the pattern of integrins expressed in MG-63 human osteosarcoma cells and causes a selective loss of cell adhesion to laminin. J Biol Chem. 1989 Dec 25;264(36):21806–21811. [PubMed] [Google Scholar]
  25. Huang M. M., Bolen J. B., Barnwell J. W., Shattil S. J., Brugge J. S. Membrane glycoprotein IV (CD36) is physically associated with the Fyn, Lyn, and Yes protein-tyrosine kinases in human platelets. Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7844–7848. doi: 10.1073/pnas.88.17.7844. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Hynes R. O. Alteration of cell-surface proteins by viral transformation and by proteolysis. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3170–3174. doi: 10.1073/pnas.70.11.3170. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Hynes R. O. Integrins: versatility, modulation, and signaling in cell adhesion. Cell. 1992 Apr 3;69(1):11–25. doi: 10.1016/0092-8674(92)90115-s. [DOI] [PubMed] [Google Scholar]
  28. Ingber D. E. Cellular tensegrity: defining new rules of biological design that govern the cytoskeleton. J Cell Sci. 1993 Mar;104(Pt 3):613–627. doi: 10.1242/jcs.104.3.613. [DOI] [PubMed] [Google Scholar]
  29. Ingber D. E., Prusty D., Frangioni J. V., Cragoe E. J., Jr, Lechene C., Schwartz M. A. Control of intracellular pH and growth by fibronectin in capillary endothelial cells. J Cell Biol. 1990 May;110(5):1803–1811. doi: 10.1083/jcb.110.5.1803. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Janat M. F., Argraves W. S., Liau G. Regulation of vascular smooth muscle cell integrin expression by transforming growth factor beta1 and by platelet-derived growth factor-BB. J Cell Physiol. 1992 Jun;151(3):588–595. doi: 10.1002/jcp.1041510319. [DOI] [PubMed] [Google Scholar]
  31. Juliano R. L., Haskill S. Signal transduction from the extracellular matrix. J Cell Biol. 1993 Feb;120(3):577–585. doi: 10.1083/jcb.120.3.577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Klein C. E., Dressel D., Steinmayer T., Mauch C., Eckes B., Krieg T., Bankert R. B., Weber L. Integrin alpha 2 beta 1 is upregulated in fibroblasts and highly aggressive melanoma cells in three-dimensional collagen lattices and mediates the reorganization of collagen I fibrils. J Cell Biol. 1991 Dec;115(5):1427–1436. doi: 10.1083/jcb.115.5.1427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Knighton D. R., Ciresi K. F., Fiegel V. D., Austin L. L., Butler E. L. Classification and treatment of chronic nonhealing wounds. Successful treatment with autologous platelet-derived wound healing factors (PDWHF). Ann Surg. 1986 Sep;204(3):322–330. doi: 10.1097/00000658-198609000-00011. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Kupper T. S., Ferguson T. A. A potential pathophysiologic role for alpha 2 beta 1 integrin in human eye diseases involving vitreoretinal traction. FASEB J. 1993 Nov;7(14):1401–1406. doi: 10.1096/fasebj.7.14.8224612. [DOI] [PubMed] [Google Scholar]
  35. Lin Y. C., Grinnell F. Decreased level of PDGF-stimulated receptor autophosphorylation by fibroblasts in mechanically relaxed collagen matrices. J Cell Biol. 1993 Aug;122(3):663–672. doi: 10.1083/jcb.122.3.663. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Marx M., Daniel T. O., Kashgarian M., Madri J. A. Spatial organization of the extracellular matrix modulates the expression of PDGF-receptor subunits in mesangial cells. Kidney Int. 1993 May;43(5):1027–1041. doi: 10.1038/ki.1993.145. [DOI] [PubMed] [Google Scholar]
  37. McNamee H. P., Ingber D. E., Schwartz M. A. Adhesion to fibronectin stimulates inositol lipid synthesis and enhances PDGF-induced inositol lipid breakdown. J Cell Biol. 1993 May;121(3):673–678. doi: 10.1083/jcb.121.3.673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Nakagawa S., Pawelek P., Grinnell F. Extracellular matrix organization modulates fibroblast growth and growth factor responsiveness. Exp Cell Res. 1989 Jun;182(2):572–582. doi: 10.1016/0014-4827(89)90260-7. [DOI] [PubMed] [Google Scholar]
  39. Nishiyama T., Akutsu N., Horii I., Nakayama Y., Ozawa T., Hayashi T. Response to growth factors of human dermal fibroblasts in a quiescent state owing to cell-matrix contact inhibition. Matrix. 1991 Apr;11(2):71–75. doi: 10.1016/s0934-8832(11)80210-6. [DOI] [PubMed] [Google Scholar]
  40. Penttinen R. P., Kobayashi S., Bornstein P. Transforming growth factor beta increases mRNA for matrix proteins both in the presence and in the absence of changes in mRNA stability. Proc Natl Acad Sci U S A. 1988 Feb;85(4):1105–1108. doi: 10.1073/pnas.85.4.1105. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Pfaff M., Aumailley M., Specks U., Knolle J., Zerwes H. G., Timpl R. Integrin and Arg-Gly-Asp dependence of cell adhesion to the native and unfolded triple helix of collagen type VI. Exp Cell Res. 1993 May;206(1):167–176. doi: 10.1006/excr.1993.1134. [DOI] [PubMed] [Google Scholar]
  42. Ross R., Bowen-Pope D. F., Raines E. W. Platelet-derived growth factor and its role in health and disease. Philos Trans R Soc Lond B Biol Sci. 1990 Mar 12;327(1239):155–169. doi: 10.1098/rstb.1990.0051. [DOI] [PubMed] [Google Scholar]
  43. Ruoslahti E. Integrins. J Clin Invest. 1991 Jan;87(1):1–5. doi: 10.1172/JCI114957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Ruoslahti E., Vaheri A. Novel human serum protein from fibroblast plasma membrane. Nature. 1974 Apr 26;248(5451):789–791. doi: 10.1038/248789a0. [DOI] [PubMed] [Google Scholar]
  45. Schmidt K. A puzzle: how similar signals yield different effects. Science. 1994 Oct 28;266(5185):566–567. doi: 10.1126/science.7939704. [DOI] [PubMed] [Google Scholar]
  46. Schwartz M. A., Lechene C., Ingber D. E. Insoluble fibronectin activates the Na/H antiporter by clustering and immobilizing integrin alpha 5 beta 1, independent of cell shape. Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7849–7853. doi: 10.1073/pnas.88.17.7849. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Shimbara N., Takashina M., Sato C., Iizuka M., Kobayashi S., Tanaka K., Ichihara A. c-myc expression is down-regulated by cell-cell and cell-extracellular matrix contacts in normal hepatocytes, but not in hepatoma cells. Biochem Biophys Res Commun. 1992 Apr 30;184(2):825–831. doi: 10.1016/0006-291x(92)90664-7. [DOI] [PubMed] [Google Scholar]
  48. Singhvi R., Kumar A., Lopez G. P., Stephanopoulos G. N., Wang D. I., Whitesides G. M., Ingber D. E. Engineering cell shape and function. Science. 1994 Apr 29;264(5159):696–698. doi: 10.1126/science.8171320. [DOI] [PubMed] [Google Scholar]
  49. Staatz W. D., Fok K. F., Zutter M. M., Adams S. P., Rodriguez B. A., Santoro S. A. Identification of a tetrapeptide recognition sequence for the alpha 2 beta 1 integrin in collagen. J Biol Chem. 1991 Apr 25;266(12):7363–7367. [PubMed] [Google Scholar]
  50. Symons M. H., Mitchison T. J. A GTPase controls cell-substrate adhesion in Xenopus XTC fibroblasts. J Cell Biol. 1992 Sep;118(5):1235–1244. doi: 10.1083/jcb.118.5.1235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Takada Y., Hemler M. E. The primary structure of the VLA-2/collagen receptor alpha 2 subunit (platelet GPIa): homology to other integrins and the presence of a possible collagen-binding domain. J Cell Biol. 1989 Jul;109(1):397–407. doi: 10.1083/jcb.109.1.397. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Takada Y., Murphy E., Pil P., Chen C., Ginsberg M. H., Hemler M. E. Molecular cloning and expression of the cDNA for alpha 3 subunit of human alpha 3 beta 1 (VLA-3), an integrin receptor for fibronectin, laminin, and collagen. J Cell Biol. 1991 Oct;115(1):257–266. doi: 10.1083/jcb.115.1.257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Takada Y., Wayner E. A., Carter W. G., Hemler M. E. Extracellular matrix receptors, ECMRII and ECMRI, for collagen and fibronectin correspond to VLA-2 and VLA-3 in the VLA family of heterodimers. J Cell Biochem. 1988 Aug;37(4):385–393. doi: 10.1002/jcb.240370406. [DOI] [PubMed] [Google Scholar]
  54. Tsuji T., Hakomori S., Osawa T. Identification of human galactoprotein b3, an oncogenic transformation-induced membrane glycoprotein, as VLA-3 alpha subunit: the primary structure of human integrin alpha 3. J Biochem. 1991 Apr;109(4):659–665. doi: 10.1093/oxfordjournals.jbchem.a123436. [DOI] [PubMed] [Google Scholar]
  55. Vandenberg P., Kern A., Ries A., Luckenbill-Edds L., Mann K., Kühn K. Characterization of a type IV collagen major cell binding site with affinity to the alpha 1 beta 1 and the alpha 2 beta 1 integrins. J Cell Biol. 1991 Jun;113(6):1475–1483. doi: 10.1083/jcb.113.6.1475. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Wayner E. A., Carter W. G. Identification of multiple cell adhesion receptors for collagen and fibronectin in human fibrosarcoma cells possessing unique alpha and common beta subunits. J Cell Biol. 1987 Oct;105(4):1873–1884. doi: 10.1083/jcb.105.4.1873. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Welch M. P., Odland G. F., Clark R. A. Temporal relationships of F-actin bundle formation, collagen and fibronectin matrix assembly, and fibronectin receptor expression to wound contraction. J Cell Biol. 1990 Jan;110(1):133–145. doi: 10.1083/jcb.110.1.133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Werb Z., Tremble P. M., Behrendtsen O., Crowley E., Damsky C. H. Signal transduction through the fibronectin receptor induces collagenase and stromelysin gene expression. J Cell Biol. 1989 Aug;109(2):877–889. doi: 10.1083/jcb.109.2.877. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Yamada K. M., Weston J. A. Isolation of a major cell surface glycoprotein from fibroblasts. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3492–3496. doi: 10.1073/pnas.71.9.3492. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Yamamoto K., Yamamoto M. Cell adhesion receptors for native and denatured type I collagens and fibronectin in rabbit arterial smooth muscle cells in culture. Exp Cell Res. 1994 Sep;214(1):258–263. doi: 10.1006/excr.1994.1256. [DOI] [PubMed] [Google Scholar]
  61. Yeo E. J., Exton J. H. Stimulation of phospholipase D by epidermal growth factor requires protein kinase C activation in Swiss 3T3 cells. J Biol Chem. 1995 Feb 24;270(8):3980–3988. doi: 10.1074/jbc.270.8.3980. [DOI] [PubMed] [Google Scholar]
  62. Zandomeni R., Bunick D., Ackerman S., Mittleman B., Weinmann R. Mechanism of action of DRB. III. Effect on specific in vitro initiation of transcription. J Mol Biol. 1983 Jul 5;167(3):561–574. doi: 10.1016/s0022-2836(83)80098-9. [DOI] [PubMed] [Google Scholar]
  63. Zaret K. S., DiPersio C. M., Jackson D. A., Montigny W. J., Weinstat D. L. Conditional enhancement of liver-specific gene transcription. Proc Natl Acad Sci U S A. 1988 Dec;85(23):9076–9080. doi: 10.1073/pnas.85.23.9076. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES