Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1993 Nov 1;123(3):749–758. doi: 10.1083/jcb.123.3.749

TGF-beta 1 stimulation of cell locomotion utilizes the hyaluronan receptor RHAMM and hyaluronan

PMCID: PMC2200130  PMID: 7693717

Abstract

TGF-beta is a potent stimulator of motility in a variety of cell types. It has recently been shown that hyaluronan (HA) can directly promote locomotion of cells through interaction with the HA receptor RHAMM. We have investigated the role of RHAMM and HA in TGF-beta-stimulated locomotion and show that TGF-beta triggers the transcription, synthesis and membrane expression of the RHAMM receptor and the secretion of HA coincident with the induction of the locomotory response. This was demonstrated by both incubating cells with exogenous TGF-beta 1 and by stimulating the production of bioactive TGF-beta 1 in tumor cells transfected with TGF-beta 1 under the control of the metallothionein promoter. TGF-beta 1-induced locomotion was suppressed by antibodies that prevented HA/RHAMM interaction, using polyclonal antibodies to either RHAMM fusion protein or RHAMM peptides, or mAbs to purified RHAMM. Peptides corresponding to the HA-binding motif of RHAMM also suppressed TGF-beta 1-induced increases in motility rate. Spontaneous locomotion of fibrosarcoma cells was blocked by neutralizing secreted TGF-beta with panspecific TGF-beta antibodies and by inhibition of TGF- beta 1 secretion with antisense oligonucleotides. Polyclonal anti-RHAMM fusion protein antibodies and peptide from the RHAMM HA-binding motif also suppressed the spontaneous motility rate of fibrosarcoma cells. These data suggest that fibrosarcoma cell locomotion requires TGF-beta, and the pathway by which TGF-beta stimulates locomotion uses the HA receptor RHAMM and HA.

Full Text

The Full Text of this article is available as a PDF (1.5 MB).

Selected References

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

  1. Allen J. B., Manthey C. L., Hand A. R., Ohura K., Ellingsworth L., Wahl S. M. Rapid onset synovial inflammation and hyperplasia induced by transforming growth factor beta. J Exp Med. 1990 Jan 1;171(1):231–247. doi: 10.1084/jem.171.1.231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anzano M. A., Roberts A. B., De Larco J. E., Wakefield L. M., Assoian R. K., Roche N. S., Smith J. M., Lazarus J. E., Sporn M. B. Increased secretion of type beta transforming growth factor accompanies viral transformation of cells. Mol Cell Biol. 1985 Jan;5(1):242–247. doi: 10.1128/mcb.5.1.242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barnard J. A., Lyons R. M., Moses H. L. The cell biology of transforming growth factor beta. Biochim Biophys Acta. 1990 Jun 1;1032(1):79–87. doi: 10.1016/0304-419x(90)90013-q. [DOI] [PubMed] [Google Scholar]
  4. Bassols A., Massagué J. Transforming growth factor beta regulates the expression and structure of extracellular matrix chondroitin/dermatan sulfate proteoglycans. J Biol Chem. 1988 Feb 25;263(6):3039–3045. [PubMed] [Google Scholar]
  5. Bray B. A., Sampson P. M., Osman M., Giandomenico A., Turino G. M. Early changes in lung tissue hyaluronan (hyaluronic acid) and hyaluronidase in bleomycin-induced alveolitis in hamsters. Am Rev Respir Dis. 1991 Feb;143(2):284–288. doi: 10.1164/ajrccm/143.2.284. [DOI] [PubMed] [Google Scholar]
  6. Chan B. M., Kassner P. D., Schiro J. A., Byers H. R., Kupper T. S., Hemler M. E. Distinct cellular functions mediated by different VLA integrin alpha subunit cytoplasmic domains. Cell. 1992 Mar 20;68(6):1051–1060. doi: 10.1016/0092-8674(92)90077-p. [DOI] [PubMed] [Google Scholar]
  7. Chen J. K., Hoshi H., McKeehan W. L. Transforming growth factor type beta specifically stimulates synthesis of proteoglycan in human adult arterial smooth muscle cells. Proc Natl Acad Sci U S A. 1987 Aug;84(15):5287–5291. doi: 10.1073/pnas.84.15.5287. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Culty M., Miyake K., Kincade P. W., Sikorski E., Butcher E. C., Underhill C., Silorski E. The hyaluronate receptor is a member of the CD44 (H-CAM) family of cell surface glycoproteins. J Cell Biol. 1990 Dec;111(6 Pt 1):2765–2774. doi: 10.1083/jcb.111.6.2765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dalal B. I., Keown P. A., Greenberg A. H. Immunocytochemical localization of secreted transforming growth factor-beta 1 to the advancing edges of primary tumors and to lymph node metastases of human mammary carcinoma. Am J Pathol. 1993 Aug;143(2):381–389. [PMC free article] [PubMed] [Google Scholar]
  10. Danielpour D., Dart L. L., Flanders K. C., Roberts A. B., Sporn M. B. Immunodetection and quantitation of the two forms of transforming growth factor-beta (TGF-beta 1 and TGF-beta 2) secreted by cells in culture. J Cell Physiol. 1989 Jan;138(1):79–86. doi: 10.1002/jcp.1041380112. [DOI] [PubMed] [Google Scholar]
  11. Delpech B., Halavent C. Characterization and purification from human brain of a hyaluronic acid-binding glycoprotein, hyaluronectin. J Neurochem. 1981 Mar;36(3):855–859. doi: 10.1111/j.1471-4159.1981.tb01672.x. [DOI] [PubMed] [Google Scholar]
  12. Derynck R., Goeddel D. V., Ullrich A., Gutterman J. U., Williams R. D., Bringman T. S., Berger W. H. Synthesis of messenger RNAs for transforming growth factors alpha and beta and the epidermal growth factor receptor by human tumors. Cancer Res. 1987 Feb 1;47(3):707–712. [PubMed] [Google Scholar]
  13. Doege K., Sasaki M., Horigan E., Hassell J. R., Yamada Y. Complete primary structure of the rat cartilage proteoglycan core protein deduced from cDNA clones. J Biol Chem. 1987 Dec 25;262(36):17757–17767. [PubMed] [Google Scholar]
  14. Faassen A. E., Schrager J. A., Klein D. J., Oegema T. R., Couchman J. R., McCarthy J. B. A cell surface chondroitin sulfate proteoglycan, immunologically related to CD44, is involved in type I collagen-mediated melanoma cell motility and invasion. J Cell Biol. 1992 Jan;116(2):521–531. doi: 10.1083/jcb.116.2.521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Fava R. A., Olsen N. J., Postlethwaite A. E., Broadley K. N., Davidson J. M., Nanney L. B., Lucas C., Townes A. S. Transforming growth factor beta 1 (TGF-beta 1) induced neutrophil recruitment to synovial tissues: implications for TGF-beta-driven synovial inflammation and hyperplasia. J Exp Med. 1991 May 1;173(5):1121–1132. doi: 10.1084/jem.173.5.1121. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Goetinck P. F., Stirpe N. S., Tsonis P. A., Carlone D. The tandemly repeated sequences of cartilage link protein contain the sites for interaction with hyaluronic acid. J Cell Biol. 1987 Nov;105(5):2403–2408. doi: 10.1083/jcb.105.5.2403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gough N. M. Rapid and quantitative preparation of cytoplasmic RNA from small numbers of cells. Anal Biochem. 1988 Aug 15;173(1):93–95. doi: 10.1016/0003-2697(88)90164-9. [DOI] [PubMed] [Google Scholar]
  18. Hardwick C., Hoare K., Owens R., Hohn H. P., Hook M., Moore D., Cripps V., Austen L., Nance D. M., Turley E. A. Molecular cloning of a novel hyaluronan receptor that mediates tumor cell motility. J Cell Biol. 1992 Jun;117(6):1343–1350. doi: 10.1083/jcb.117.6.1343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Heldin P., Laurent T. C., Heldin C. H. Effect of growth factors on hyaluronan synthesis in cultured human fibroblasts. Biochem J. 1989 Mar 15;258(3):919–922. doi: 10.1042/bj2580919. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hurta R. A., Samuel S. K., Greenberg A. H., Wright J. A. Early induction of ribonucleotide reductase gene expression by transforming growth factor beta 1 in malignant H-ras transformed cell lines. J Biol Chem. 1991 Dec 15;266(35):24097–24100. [PubMed] [Google Scholar]
  22. 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]
  23. Hök M., Kjellén L., Johansson S. Cell-surface glycosaminoglycans. Annu Rev Biochem. 1984;53:847–869. doi: 10.1146/annurev.bi.53.070184.004215. [DOI] [PubMed] [Google Scholar]
  24. Khalil N., Bereznay O., Sporn M., Greenberg A. H. Macrophage production of transforming growth factor beta and fibroblast collagen synthesis in chronic pulmonary inflammation. J Exp Med. 1989 Sep 1;170(3):727–737. doi: 10.1084/jem.170.3.727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Khalil N., Greenberg A. H. The role of TGF-beta in pulmonary fibrosis. Ciba Found Symp. 1991;157:194–211. doi: 10.1002/9780470514061.ch13. [DOI] [PubMed] [Google Scholar]
  26. Kimata K., Honma Y., Okayama M., Oguri K., Hozumi M., Suzuki S. Increased synthesis of hyaluronic acid by mouse mammary carcinoma cell variants with high metastatic potential. Cancer Res. 1983 Mar;43(3):1347–1354. [PubMed] [Google Scholar]
  27. Klein-Soyer C., Beretz A., Cazenave J. P., Wittendorp-Rechenmann E., Vonesch J. L., Rechenmann R. V., Driot F., Maffrand J. P. Sulfated polysaccharides modulate effects of acidic and basic fibroblast growth factors on repair of injured confluent human vascular endothelium. Arteriosclerosis. 1989 Mar-Apr;9(2):147–153. doi: 10.1161/01.atv.9.2.147. [DOI] [PubMed] [Google Scholar]
  28. Krusius T., Gehlsen K. R., Ruoslahti E. A fibroblast chondroitin sulfate proteoglycan core protein contains lectin-like and growth factor-like sequences. J Biol Chem. 1987 Sep 25;262(27):13120–13125. [PubMed] [Google Scholar]
  29. Kähäri V. M., Larjava H., Uitto J. Differential regulation of extracellular matrix proteoglycan (PG) gene expression. Transforming growth factor-beta 1 up-regulates biglycan (PGI), and versican (large fibroblast PG) but down-regulates decorin (PGII) mRNA levels in human fibroblasts in culture. J Biol Chem. 1991 Jun 5;266(16):10608–10615. [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. Liotta L. A., Schiffmann E. Tumour motility factors. Cancer Surv. 1988;7(4):631–652. [PubMed] [Google Scholar]
  32. Madri J. A., Pratt B. M., Tucker A. M. Phenotypic modulation of endothelial cells by transforming growth factor-beta depends upon the composition and organization of the extracellular matrix. J Cell Biol. 1988 Apr;106(4):1375–1384. doi: 10.1083/jcb.106.4.1375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. McClarty G. A., Chan A. K., Choy B. K., Wright J. A. Reversion of hydroxyurea resistance, decline in ribonucleotide reductase activity, and loss of M2 gene amplification. Biochem Biophys Res Commun. 1987 Jun 30;145(3):1276–1282. doi: 10.1016/0006-291x(87)91575-0. [DOI] [PubMed] [Google Scholar]
  34. Mooradian D. L., McCarthy J. B., Komanduri K. V., Furcht L. T. Effects of transforming growth factor-beta 1 on human pulmonary adenocarcinoma cell adhesion, motility, and invasion in vitro. J Natl Cancer Inst. 1992 Apr 1;84(7):523–527. doi: 10.1093/jnci/84.7.523. [DOI] [PubMed] [Google Scholar]
  35. Neame P. J., Christner J. E., Baker J. R. The primary structure of link protein from rat chondrosarcoma proteoglycan aggregate. J Biol Chem. 1986 Mar 15;261(8):3519–3535. [PubMed] [Google Scholar]
  36. Nettelbladt O., Bergh J., Schenholm M., Tengblad A., Hällgren R. Accumulation of hyaluronic acid in the alveolar interstitial tissue in bleomycin-induced alveolitis. Am Rev Respir Dis. 1989 Mar;139(3):759–762. doi: 10.1164/ajrccm/139.3.759. [DOI] [PubMed] [Google Scholar]
  37. Nugent M. A., Edelman E. R. Transforming growth factor beta 1 stimulates the production of basic fibroblast growth factor binding proteoglycans in Balb/c3T3 cells. J Biol Chem. 1992 Oct 15;267(29):21256–21264. [PubMed] [Google Scholar]
  38. Partin A. W., Isaacs J. T., Treiger B., Coffey D. S. Early cell motility changes associated with an increase in metastatic ability in rat prostatic cancer cells transfected with the v-Harvey-ras oncogene. Cancer Res. 1988 Nov 1;48(21):6050–6053. [PubMed] [Google Scholar]
  39. Partin A. W., Schoeniger J. S., Mohler J. L., Coffey D. S. Fourier analysis of cell motility: correlation of motility with metastatic potential. Proc Natl Acad Sci U S A. 1989 Feb;86(4):1254–1258. doi: 10.1073/pnas.86.4.1254. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Perides G., Lane W. S., Andrews D., Dahl D., Bignami A. Isolation and partial characterization of a glial hyaluronate-binding protein. J Biol Chem. 1989 Apr 5;264(10):5981–5987. [PubMed] [Google Scholar]
  41. Perrotti D., Cimino L., Ferrari S., Sacchi A. Differential expression of transforming growth factor-beta 1 gene in 3LL metastatic variants. Cancer Res. 1991 Oct 15;51(20):5491–5494. [PubMed] [Google Scholar]
  42. Pierce G. F., Mustoe T. A., Lingelbach J., Masakowski V. R., Gramates P., Deuel T. F. Transforming growth factor beta reverses the glucocorticoid-induced wound-healing deficit in rats: possible regulation in macrophages by platelet-derived growth factor. Proc Natl Acad Sci U S A. 1989 Apr;86(7):2229–2233. doi: 10.1073/pnas.86.7.2229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Postlethwaite A. E., Keski-Oja J., Moses H. L., Kang A. H. Stimulation of the chemotactic migration of human fibroblasts by transforming growth factor beta. J Exp Med. 1987 Jan 1;165(1):251–256. doi: 10.1084/jem.165.1.251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Reibman J., Meixler S., Lee T. C., Gold L. I., Cronstein B. N., Haines K. A., Kolasinski S. L., Weissmann G. Transforming growth factor beta 1, a potent chemoattractant for human neutrophils, bypasses classic signal-transduction pathways. Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6805–6809. doi: 10.1073/pnas.88.15.6805. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Samuel S. K., Hurta R. A., Kondaiah P., Khalil N., Turley E. A., Wright J. A., Greenberg A. H. Autocrine induction of tumor protease production and invasion by a metallothionein-regulated TGF-beta 1 (Ser223, 225). EMBO J. 1992 Apr;11(4):1599–1605. doi: 10.1002/j.1460-2075.1992.tb05205.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Sato Y., Rifkin D. B. Autocrine activities of basic fibroblast growth factor: regulation of endothelial cell movement, plasminogen activator synthesis, and DNA synthesis. J Cell Biol. 1988 Sep;107(3):1199–1205. doi: 10.1083/jcb.107.3.1199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Schor S. L., Schor A. M., Grey A. M., Chen J., Rushton G., Grant M. E., Ellis I. Mechanism of action of the migration stimulating factor produced by fetal and cancer patient fibroblasts: effect on hyaluronic and synthesis. In Vitro Cell Dev Biol. 1989 Aug;25(8):737–746. doi: 10.1007/BF02623727. [DOI] [PubMed] [Google Scholar]
  48. Schwarz L. C., Wright J. A., Gingras M. C., Kondaiah P., Danielpour D., Pimentel M., Sporn M. B., Greenberg A. H. Aberrant TGF-beta production and regulation in metastatic malignancy. Growth Factors. 1990;3(2):115–127. doi: 10.3109/08977199009108274. [DOI] [PubMed] [Google Scholar]
  49. Stamenkovic I., Aruffo A., Amiot M., Seed B. The hematopoietic and epithelial forms of CD44 are distinct polypeptides with different adhesion potentials for hyaluronate-bearing cells. EMBO J. 1991 Feb;10(2):343–348. doi: 10.1002/j.1460-2075.1991.tb07955.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Stoker M., Gherardi E. Regulation of cell movement: the motogenic cytokines. Biochim Biophys Acta. 1991 Apr 16;1072(1):81–102. doi: 10.1016/0304-419x(91)90008-9. [DOI] [PubMed] [Google Scholar]
  51. Thomas L., Byers H. R., Vink J., Stamenkovic I. CD44H regulates tumor cell migration on hyaluronate-coated substrate. J Cell Biol. 1992 Aug;118(4):971–977. doi: 10.1083/jcb.118.4.971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Toole B. P., Biswas C., Gross J. Hyaluronate and invasiveness of the rabbit V2 carcinoma. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6299–6303. doi: 10.1073/pnas.76.12.6299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Toole B. P. Hyaluronan and its binding proteins, the hyaladherins. Curr Opin Cell Biol. 1990 Oct;2(5):839–844. doi: 10.1016/0955-0674(90)90081-o. [DOI] [PubMed] [Google Scholar]
  54. Toole B. P., Munaim S. I., Welles S., Knudson C. B. Hyaluronate-cell interactions and growth factor regulation of hyaluronate synthesis during limb development. Ciba Found Symp. 1989;143:138–285. doi: 10.1002/9780470513774.ch9. [DOI] [PubMed] [Google Scholar]
  55. Turley E. A., Austen L., Vandeligt K., Clary C. Hyaluronan and a cell-associated hyaluronan binding protein regulate the locomotion of ras-transformed cells. J Cell Biol. 1991 Mar;112(5):1041–1047. doi: 10.1083/jcb.112.5.1041. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Turley E. A. Hyaluronan and cell locomotion. Cancer Metastasis Rev. 1992 Mar;11(1):21–30. doi: 10.1007/BF00047600. [DOI] [PubMed] [Google Scholar]
  57. Turley E. A., Moore D., Hayden L. J. Characterization of hyaluronate binding proteins isolated from 3T3 and murine sarcoma virus transformed 3T3 cells. Biochemistry. 1987 Jun 2;26(11):2997–3005. doi: 10.1021/bi00385a007. [DOI] [PubMed] [Google Scholar]
  58. Turley E. A., Torrance J. Localization of hyaluronate and hyaluronate-binding protein on motile and non-motile fibroblasts. Exp Cell Res. 1985 Nov;161(1):17–28. doi: 10.1016/0014-4827(85)90486-0. [DOI] [PubMed] [Google Scholar]
  59. Turley E. A., Tretiak M. Glycosaminoglycan production by murine melanoma variants in vivo and in vitro. Cancer Res. 1985 Oct;45(10):5098–5105. [PubMed] [Google Scholar]
  60. Turley E., Auersperg N. A hyaluronate binding protein transiently codistributes with p21k-ras in cultured cell lines. Exp Cell Res. 1989 Jun;182(2):340–348. doi: 10.1016/0014-4827(89)90239-5. [DOI] [PubMed] [Google Scholar]
  61. Wahl S. M., Hunt D. A., Wakefield L. M., McCartney-Francis N., Wahl L. M., Roberts A. B., Sporn M. B. Transforming growth factor type beta induces monocyte chemotaxis and growth factor production. Proc Natl Acad Sci U S A. 1987 Aug;84(16):5788–5792. doi: 10.1073/pnas.84.16.5788. [DOI] [PMC free article] [PubMed] [Google Scholar]
  62. Welch D. R., Fabra A., Nakajima M. Transforming growth factor beta stimulates mammary adenocarcinoma cell invasion and metastatic potential. Proc Natl Acad Sci U S A. 1990 Oct;87(19):7678–7682. doi: 10.1073/pnas.87.19.7678. [DOI] [PMC free article] [PubMed] [Google Scholar]
  63. Yamada K. M., Kennedy D. W., Yamada S. S., Gralnick H., Chen W. T., Akiyama S. K. Monoclonal antibody and synthetic peptide inhibitors of human tumor cell migration. Cancer Res. 1990 Aug 1;50(15):4485–4496. [PubMed] [Google Scholar]
  64. Yamaguchi Y., Mann D. M., Ruoslahti E. Negative regulation of transforming growth factor-beta by the proteoglycan decorin. Nature. 1990 Jul 19;346(6281):281–284. doi: 10.1038/346281a0. [DOI] [PubMed] [Google Scholar]
  65. Yang B., Zhang L., Turley E. A. Identification of two hyaluronan-binding domains in the hyaluronan receptor RHAMM. J Biol Chem. 1993 Apr 25;268(12):8617–8623. [PubMed] [Google Scholar]

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

RESOURCES