Skip to main content
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1987 Mar 1;104(3):601–610. doi: 10.1083/jcb.104.3.601

Induction of fibronectin matrix assembly in human fibrosarcoma cells by dexamethasone

PMCID: PMC2114531  PMID: 2950120

Abstract

Previous studies have suggested that the assembly of fibronectin into the extracellular matrix of cultured fibroblasts is mediated by specific matrix assembly receptors that recognize a binding site in the amino terminus of the fibronectin molecule (McKeown-Longo, P.J., and D.F. Mosher, 1985, J. Cell Biol., 100:364-374). In the presence of dexamethasone, human fibrosarcoma cells (HT-1080) acquired the ability to specifically bind exogenous plasma fibronectin and incorporate it into a detergent-insoluble extracellular matrix. Dexamethasone-induced fibronectin binding to HT-1080 cells was time dependent, dose dependent, and inhibited by cycloheximide. Saturation binding curves indicated that dexamethasone induced the appearance of 7.7 X 10(4) matrix assembly receptors per cell. The induced receptors exhibited a dissociation constant (KD) for soluble fibronectin of 5.0 X 10(-8) M. In parallel experiments, normal fibroblasts exhibited 4.1 X 10(5) receptors (KD = 5.3 X 10(-8) M) per cell. In the presence of cycloheximide, the induced fibronectin-binding activity on HT-1080 cells returned to uninduced levels within 12 h. In contrast, fibronectin-binding activity on normal fibroblasts was stable in the presence of cycloheximide for up to 54 h. The first-order rate constant (Kt = 2.07 X 10(-4) min-1) for the transfer of receptor-bound fibronectin to extracellular matrix was four- to fivefold less than that for normal fibroblasts (Kt = 1.32 X 10(-3) min-1). Lactoperoxidase- catalyzed iodination of HT-1080 monolayers indicated that a 48,000-mol- wt cell surface protein was enhanced with dexamethasone. The results from these experiments suggest that dexamethasone induces functional matrix assembly receptors on the surface of HT-1080 cells; however, the rate of incorporation of fibronectin into the matrix is much slower than that of normal fibroblasts.

Full Text

The Full Text of this article is available as a PDF (1.9 MB).

Selected References

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

  1. Akiyama S. K., Yamada S. S., Yamada K. M. Characterization of a 140-kD avian cell surface antigen as a fibronectin-binding molecule. J Cell Biol. 1986 Feb;102(2):442–448. doi: 10.1083/jcb.102.2.442. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ali I. U., Mautner V., Lanza R., Hynes R. O. Restoration of normal morphology, adhesion and cytoskeleton in transformed cells by addition of a transformation-sensitive surface protein. Cell. 1977 May;11(1):115–126. doi: 10.1016/0092-8674(77)90322-1. [DOI] [PubMed] [Google Scholar]
  3. Alitalo K., Keski-Oja J., Hedman K., Vaheri A. Loss of different pericellular matrix components of rat cells transformed with a T-class ts mutant of Rous sarcoma virus. Virology. 1982 Jun;119(2):347–357. doi: 10.1016/0042-6822(82)90094-0. [DOI] [PubMed] [Google Scholar]
  4. Alitalo K., Vaheri A. Pericellular matrix in malignant transformation. Adv Cancer Res. 1982;37:111–158. doi: 10.1016/s0065-230x(08)60883-0. [DOI] [PubMed] [Google Scholar]
  5. Aplin J. D., Hughes R. C., Jaffe C. L., Sharon N. Reversible cross-linking of cellular components of adherent fibroblasts to fibronectin and lectin-coated substrata. Exp Cell Res. 1981 Aug;134(2):488–494. doi: 10.1016/0014-4827(81)90453-5. [DOI] [PubMed] [Google Scholar]
  6. Armelin M. C., Armelin H. A. Glucocorticoid hormone modulation of both cell surface and cytoskeleton related to growth control of rat glioma cells. J Cell Biol. 1983 Aug;97(2):459–465. doi: 10.1083/jcb.97.2.459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Baumann H., Eldredge D. Dexamethasone increases the synthesis and secretion of a partially active fibronectin in rat hepatoma cells. J Cell Biol. 1982 Oct;95(1):29–40. doi: 10.1083/jcb.95.1.29. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chen L. B., Murray A., Segal R. A., Bushnell A., Walsh M. L. Studies on intercellular LETS glycoprotein matrices. Cell. 1978 Jun;14(2):377–391. doi: 10.1016/0092-8674(78)90123-x. [DOI] [PubMed] [Google Scholar]
  9. Cwikel B. J., Barouski-Miller P. A., Coleman P. L., Gelehrter T. D. Dexamethasone induction of an inhibitor of plasminogen activator in HTC hepatoma cells. J Biol Chem. 1984 Jun 10;259(11):6847–6851. [PubMed] [Google Scholar]
  10. Fagan J. B., Sobel M. E., Yamada K. M., de Crombrugghe B., Pastan I. Effects of transformation on fibronectin gene expression using cloned fibronectin cDNA. J Biol Chem. 1981 Jan 10;256(1):520–525. [PubMed] [Google Scholar]
  11. Furcht L. T., Mosher D. F., Wendelschafer-Crabb G., Foidart J. M. Reversal by glucocorticoid hormones of the loss of a fibronectin and probollagen matrix around transformed human cells. Cancer Res. 1979 Jun;39(6 Pt 1):2077–2083. [PubMed] [Google Scholar]
  12. Furcht L. T., Mosher D. F., Wendelschafer-Crabb G., Woodbridge P. A., Foidart J. M. Dexamethasone-induced accumulation of a fibronectin and collagen extracellular matrix in transformed human cells. Nature. 1979 Feb 1;277(5695):393–395. doi: 10.1038/277393a0. [DOI] [PubMed] [Google Scholar]
  13. Furcht L. T., Smith D., Wendelschafer-Crabb G., Mosher D. F., Foidart J. M. Fibronectin presence in native collagen fibrils of human fibroblasts: immunoperoxidase and immunoferritin localization. J Histochem Cytochem. 1980 Dec;28(12):1319–1333. doi: 10.1177/28.12.7014712. [DOI] [PubMed] [Google Scholar]
  14. Hayman E. G., Engvall E., Ruoslahti E. Concomitant loss of cell surface fibronectin and laminin from transformed rat kidney cells. J Cell Biol. 1981 Feb;88(2):352–357. doi: 10.1083/jcb.88.2.352. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hayman E. G., Ruoslahti E. Distribution of fetal bovine serum fibronectin and endogenous rat cell fibronectin in extracellular matrix. J Cell Biol. 1979 Oct;83(1):255–259. doi: 10.1083/jcb.83.1.255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hedman K., Johansson S., Vartio T., Kjellén L., Vaheri A., Hök M. Structure of the pericellular matrix: association of heparan and chondroitin sulfates with fibronectin-procollagen fibers. Cell. 1982 Mar;28(3):663–671. doi: 10.1016/0092-8674(82)90221-5. [DOI] [PubMed] [Google Scholar]
  17. Hedman K., Kurkinen M., Alitalo K., Vaheri A., Johansson S., Hök M. Isolation of the pericellular matrix of human fibroblast cultures. J Cell Biol. 1979 Apr;81(1):83–91. doi: 10.1083/jcb.81.1.83. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Heller-Harrison R. A., Carter W. G. Pepsin-generated type VI collagen is a degradation product of GP140. J Biol Chem. 1984 Jun 10;259(11):6858–6864. [PubMed] [Google Scholar]
  19. Horwitz A., Duggan K., Greggs R., Decker C., Buck C. The cell substrate attachment (CSAT) antigen has properties of a receptor for laminin and fibronectin. J Cell Biol. 1985 Dec;101(6):2134–2144. doi: 10.1083/jcb.101.6.2134. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hughes R. C., Butters T. D., Aplin J. D. Cell surface molecules involved in fibronectin-mediated adhesion. A study using specific antisera. Eur J Cell Biol. 1981 Dec;26(1):198–207. [PubMed] [Google Scholar]
  21. Hynes R. O., Ali I. U., Destree A. T., Mautner V., Perkins M. E., Senger D. R., Wagner D. D., Smith K. K. A large glycoprotein lost from the surfaces of transformed cells. Ann N Y Acad Sci. 1978 Jun 20;312:317–342. doi: 10.1111/j.1749-6632.1978.tb16811.x. [DOI] [PubMed] [Google Scholar]
  22. Hynes R. O., Bye J. M. Density and cell cycle dependence of cell surface proteins in hamster fibroblasts. Cell. 1974 Oct;3(2):113–120. doi: 10.1016/0092-8674(74)90114-7. [DOI] [PubMed] [Google Scholar]
  23. Hynes R. O., Yamada K. M. Fibronectins: multifunctional modular glycoproteins. J Cell Biol. 1982 Nov;95(2 Pt 1):369–377. doi: 10.1083/jcb.95.2.369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Irish P. S., Hasty D. L. Immunocytochemical localization of fibronectin in human fibroblast cultures using a cell surface replica technique. J Histochem Cytochem. 1983 Jan;31(1):69–77. doi: 10.1177/31.1.6833740. [DOI] [PubMed] [Google Scholar]
  25. Jaffe E. A., Ruggiero J. T., Leung L. K., Doyle M. J., McKeown-Longo P. J., Mosher D. F. Cultured human fibroblasts synthesize and secrete thrombospondin and incorporate it into extracellular matrix. Proc Natl Acad Sci U S A. 1983 Feb;80(4):998–1002. doi: 10.1073/pnas.80.4.998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Jilek F., Hörmann H. Cold-insoluble globulin, II. Plasminolysis of cold-insoluble globulin. Hoppe Seylers Z Physiol Chem. 1977 Jan;358(1):133–136. [PubMed] [Google Scholar]
  27. Johansson S., Hök M. Substrate adhesion of rat hepatocytes: on the mechanism of attachment to fibronectin. J Cell Biol. 1984 Mar;98(3):810–817. doi: 10.1083/jcb.98.3.810. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Jones P. A., Laug W. E., Benedict W. F. Fibrinolytic activity in a human fibrosarcoma cell line and evidence for the induction of plasminogen activator secretion during tumor formation. Cell. 1975 Oct;6(2):245–252. doi: 10.1016/0092-8674(75)90015-x. [DOI] [PubMed] [Google Scholar]
  29. Kleinman H. K., Martin G. R., Fishman P. H. Ganglioside inhibition of fibronectin-mediated cell adhesion to collagen. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3367–3371. doi: 10.1073/pnas.76.7.3367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  31. Lehto V. P., Virtanen I. Formation of stress fibres and focal adhesion sites in monensin-exposed cultured human fibroblasts in response to exogenously added cellular fibronectin. Exp Cell Res. 1985 Jun;158(2):563–569. doi: 10.1016/0014-4827(85)90481-1. [DOI] [PubMed] [Google Scholar]
  32. Lien Y. H., Wong M. J., Golbus M. S., Stern R. Hydrocortisone stimulates fibronectin synthesis in cultured fibroblasts. J Cell Physiol. 1984 Jul;120(1):103–107. doi: 10.1002/jcp.1041200114. [DOI] [PubMed] [Google Scholar]
  33. Lyubimov A. V., Vasiliev J. M. Distribution of fibronectin-containing structures on the surface of lamelloplasm and endoplasm of fibroblasts ; hypothesis of receptor-mediated assembly of fibronectin structures. Cell Biol Int Rep. 1982 Feb;6(2):105–112. doi: 10.1016/0309-1651(82)90086-8. [DOI] [PubMed] [Google Scholar]
  34. Marceau N., Goyette R., Valet J. P., Deschenes J. The effect of dexamethasone on formation of a fibronectin extracellular matrix by rat hepatocytes in vitro. Exp Cell Res. 1980 Feb;125(2):497–502. doi: 10.1016/0014-4827(80)90146-9. [DOI] [PubMed] [Google Scholar]
  35. McKeown-Longo P. J., Mosher D. F. Binding of plasma fibronectin to cell layers of human skin fibroblasts. J Cell Biol. 1983 Aug;97(2):466–472. doi: 10.1083/jcb.97.2.466. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. McKeown-Longo P. J., Mosher D. F. Interaction of the 70,000-mol-wt amino-terminal fragment of fibronectin with the matrix-assembly receptor of fibroblasts. J Cell Biol. 1985 Feb;100(2):364–374. doi: 10.1083/jcb.100.2.364. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. McKeown-Longo P. J., Mosher D. F. Mechanism of formation of disulfide-bonded multimers of plasma fibronectin in cell layers of cultured human fibroblasts. J Biol Chem. 1984 Oct 10;259(19):12210–12215. [PubMed] [Google Scholar]
  38. Mosher D. F., McKeown-Longo P. J. Assembly of fibronectin-containing extracellular matrix: a glimpse of the machinery. Biopolymers. 1985 Jan;24(1):199–210. doi: 10.1002/bip.360240116. [DOI] [PubMed] [Google Scholar]
  39. Olden K., Yamada K. M. Mechanism of the decrease in the major cell surface protein of chick embryo fibroblasts after transformation. Cell. 1977 Aug;11(4):957–969. doi: 10.1016/0092-8674(77)90307-5. [DOI] [PubMed] [Google Scholar]
  40. Oliver N., Newby R. F., Furcht L. T., Bourgeois S. Regulation of fibronectin biosynthesis by glucocorticoids in human fibrosarcoma cells and normal fibroblasts. Cell. 1983 May;33(1):287–296. doi: 10.1016/0092-8674(83)90357-4. [DOI] [PubMed] [Google Scholar]
  41. Oppenheimer-Marks N., Grinnell F. Calcium ions protect cell-substratum adhesion receptors against proteolysis. Evidence from immunoabsorption and electroblotting studies. Exp Cell Res. 1984 Jun;152(2):467–475. doi: 10.1016/0014-4827(84)90648-7. [DOI] [PubMed] [Google Scholar]
  42. Owens M. R., Cimino C. D. Synthesis of fibronectin by the isolated perfused rat liver. Blood. 1982 Jun;59(6):1305–1309. [PubMed] [Google Scholar]
  43. Perkins M. E., Ji T. H., Hynes R. O. Cross-linking of fibronectin to sulfated proteoglycans at the cell surface. Cell. 1979 Apr;16(4):941–952. doi: 10.1016/0092-8674(79)90109-0. [DOI] [PubMed] [Google Scholar]
  44. Pytela R., Pierschbacher M. D., Ruoslahti E. Identification and isolation of a 140 kd cell surface glycoprotein with properties expected of a fibronectin receptor. Cell. 1985 Jan;40(1):191–198. doi: 10.1016/0092-8674(85)90322-8. [DOI] [PubMed] [Google Scholar]
  45. Raugi G. J., Mumby S. M., Abbott-Brown D., Bornstein P. Thrombospondin: synthesis and secretion by cells in culture. J Cell Biol. 1982 Oct;95(1):351–354. doi: 10.1083/jcb.95.1.351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Rennard S. I., Wind M. L., Hewitt A. T., Kleinman H. K. Effect of collagen and cell shape on binding of fibronectin to cells. Arch Biochem Biophys. 1981 Jan;206(1):205–212. doi: 10.1016/0003-9861(81)90082-5. [DOI] [PubMed] [Google Scholar]
  47. Salhanick A. I., Krupp M. N., Amatruda J. M. Dexamethasone stimulates insulin receptor synthesis in cultured rat hepatocytes. J Biol Chem. 1983 Dec 10;258(23):14130–14135. [PubMed] [Google Scholar]
  48. Singer I. I. The fibronexus: a transmembrane association of fibronectin-containing fibers and bundles of 5 nm microfilaments in hamster and human fibroblasts. Cell. 1979 Mar;16(3):675–685. doi: 10.1016/0092-8674(79)90040-0. [DOI] [PubMed] [Google Scholar]
  49. Spiegel S., Schlessinger J., Fishman P. H. Incorporation of fluorescent gangliosides into human fibroblasts: mobility, fate, and interaction with fibronectin. J Cell Biol. 1984 Aug;99(2):699–704. doi: 10.1083/jcb.99.2.699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Spiegel S., Yamada K. M., Hom B. E., Moss J., Fishman P. H. Fibrillar organization of fibronectin is expressed coordinately with cell surface gangliosides in a variant murine fibroblast. J Cell Biol. 1986 May;102(5):1898–1906. doi: 10.1083/jcb.102.5.1898. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Thompson L. K., Horowitz P. M., Bentley K. L., Thomas D. D., Alderete J. F., Klebe R. J. Localization of the ganglioside-binding site of fibronectin. J Biol Chem. 1986 Apr 15;261(11):5209–5214. [PubMed] [Google Scholar]
  52. Urushihara H., Yamada K. M. Evidence for involvement of more than one class of glycoprotein in cell interactions with fibronectin. J Cell Physiol. 1986 Mar;126(3):323–332. doi: 10.1002/jcp.1041260302. [DOI] [PubMed] [Google Scholar]
  53. Wagner D. D., Ivatt R., Destree A. T., Hynes R. O. Similarities and differences between the fibronectins of normal and transformed hamster cells. J Biol Chem. 1981 Nov 25;256(22):11708–11715. [PubMed] [Google Scholar]
  54. Wiley H. S., Cunningham D. D. The endocytotic rate constant. A cellular parameter for quantitating receptor-mediated endocytosis. J Biol Chem. 1982 Apr 25;257(8):4222–4229. [PubMed] [Google Scholar]
  55. Yamada K. M. Cell surface interactions with extracellular materials. Annu Rev Biochem. 1983;52:761–799. doi: 10.1146/annurev.bi.52.070183.003553. [DOI] [PubMed] [Google Scholar]
  56. Yamada K. M., Critchley D. R., Fishman P. H., Moss J. Exogenous gangliosides enhance the interaction of fibronectin with ganglioside-deficient cells. Exp Cell Res. 1983 Feb;143(2):295–302. doi: 10.1016/0014-4827(83)90054-x. [DOI] [PubMed] [Google Scholar]
  57. Yamada K. M., Yamada S. S., Pastan I. Cell surface protein partially restores morphology, adhesiveness, and contact inhibition of movement to transformed fibroblasts. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1217–1221. doi: 10.1073/pnas.73.4.1217. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES