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Analysis of TGFβ-Mediated Synthesis of Extracellular Matrix Components

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Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 142))

Abstract

Coordinated regulation of production and turnover of extracellular matrix (ECM) components is essential for normal tissue homeostasis. The composition of the extracellular matrix can influence cell growth, state of differentiation, and specific gene induction. The multifunctional cytokine transforming growth factor-β (TGF-β) exerts its effects on cell proliferation, differentiation, and migration in part through its modulation of extracellular matrix components. TGF-β promotes net matrix deposition by increasing the expression of specific ECM components such as fibronectin (FN) and collagen, upregulating the expression of inhibitors of ECM proteases, such as plasminogen activator inhibitor-1 (PAI-1) and tissue inhibitors of matrix metaUoproteinases(TIMPs). while simultaneously suppressing the synthesis of proteases, which degrade matrix components, such as interstitial collagenase (reviewed in refs. 1 and 2). TGF-β also induces the expression of cell surface receptors for the ECM, the integrins, which enhance cell-matrix interactions (1,2).

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References

  1. Massague J. (1990) The transforming growth factor-β family. Anna. Rev. CeU Biol. 6, 597–641.

    Article  CAS  Google Scholar 

  2. Roberts A. B. and Sporn M. B. (1990) The transforming growth factor-βs, in Pep tide Growth Factors and Their Receptors (Sporn M. and Roberts A. B., eds.). Springer-Verlag, Heidelberg, pp. 419–472.

    Google Scholar 

  3. Kornblihtt A. R., Pesce C. G., Alonso C. R., Cramer P., Srebrow A., Werbajh S., and Muro A. F. (1996) The fibronectin gene as a model for splicing and transcription studies. FASEB. J. 10, 248–257.

    PubMed  CAS  Google Scholar 

  4. Hynes R. (1985) Molecular biology of fibronectin. Anna. Rev. Cell Biol. 1, 67–90.

    Article  CAS  Google Scholar 

  5. Ignotz R. A. and Massague J. (1986) Transforming growth factor-β stimulates the expression of fibronectin and collagen and their incorporation into the extracellular matrix. J. Biol. Chem. 261, 4337–4345.

    PubMed  CAS  Google Scholar 

  6. Dean D. C, Newby R. F., and Bourgeois S. (1988) Regulation of fibronectin biosynthesis by dexamethasone, transforming growth factor beta, and cAMP in human cell lines. J. CeU Biol. 106, 2159–2170.

    Article  CAS  Google Scholar 

  7. Loskutoff D. J., Sawdey M., Keeton M., and Schneiderman J. (1993) Regulation of PAI-1 gene expression in vivo. Thromb. Haemost. 70, 135–137.

    PubMed  CAS  Google Scholar 

  8. Deng G., Cuiriden S. A., Wang S., Rosenberg S., and Loskutoff D. J. (1996) Is plasminogen activator inhibitor-1 the molecular switch that governs urokinase receptor-mediated cell adhesion and release? J. Cell Biol. 134, 1563–1571.

    Article  PubMed  CAS  Google Scholar 

  9. Stefansson S., and Lawrence D. A. (1996) The serpin PAI-1 inhibits cell migration by blocking integrin avβ3 binding to vitronectin. Nature 383, 441–443.

    Article  PubMed  CAS  Google Scholar 

  10. Westerhausen D. R., Jr., Hopkins W. E., and Billadello J. J. (1991) Multiple transforming growth factor-β-indueible elements regulate expression of the plasminogen activator inhibitor type-1 gene in HepG2 cells. J. Biol. Chem. 266, 1092–1100.

    PubMed  CAS  Google Scholar 

  11. Matrisian L. M. (1990) Metalloproteinases and their inhibitors in matrix remodeling. Trends Genet. 6, 121–125.

    Article  PubMed  CAS  Google Scholar 

  12. Mauviel A., Chung K.-Y., Agarwal A., Tamai K., and Uitto J. (1996) Cellspecific induction of distinct oncogenes of the Jun family is responsible for differential regulation of collagenase gene expression by transforming growth factor β in fibroblasts and keratinocytes. J. Biol. Chem. 271, 10,917–10,923.

    Article  PubMed  CAS  Google Scholar 

  13. Kerr L. D., Miller D. B., and Matrisian L. M. (1990) TGF-beta inhibition of transin/stromelysin expression is mediated through a Fos binding sequence. Cell 61, 267–278.

    Article  PubMed  CAS  Google Scholar 

  14. Kubota S., Fridman R., and Yamada Y. (1991) Transforming growth factor-β suppresses the invasiveness of human fibrosarcoma cells in vitro by increasing expression of tissue inhibitor of metalloprotease. Biochem. Biophys. Res. Commun. 176, 129–136.

    Article  PubMed  CAS  Google Scholar 

  15. Salo T., Lyons J. G., Rahemtulla F., Birkedal-Hansen H., and Larjava H. (1991) Transforming growth factor-βl up-regulates type IV collagenase expression in cultured human keratinocytes., J. BioL Chem. 266, 11,436–11,441.

    PubMed  CAS  Google Scholar 

  16. Border W. A. and Noble N. A. (1994) Transforming growth factor (3 in tissue fibrosis. N. Engl J. Med. 331, 1286–1292.

    Article  PubMed  CAS  Google Scholar 

  17. Hocevar B. A. and Howe P. H. (1996) Isolation and characterization of mutant cell lines defective in transforming growth factor beta signaling. Proc. Natl.Acad. Sci. USA 93, 7655–7660.

    Article  PubMed  CAS  Google Scholar 

  18. Shapiro S. D., Fliszar C. J., Broekelmann T. J., Mecham R. P., Senior R. M. and Welgus H. G. (1995) Activation of the 92-kDagelatinase by stromelysin and 4-aminophenylmercuric acetate. J. Bioi. Chem. 270, 6351–6356.

    Article  CAS  Google Scholar 

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© 2000 Humana Press Inc.

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Hocevar, B.A., Howe, P.H. (2000). Analysis of TGFβ-Mediated Synthesis of Extracellular Matrix Components. In: Howe, P.H. (eds) Transforming Growth Factor-Beta Protocols. Methods in Molecular Biology™, vol 142. Humana Press. https://doi.org/10.1385/1-59259-053-5:55

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  • DOI: https://doi.org/10.1385/1-59259-053-5:55

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-646-8

  • Online ISBN: 978-1-59259-053-7

  • eBook Packages: Springer Protocols

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