Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1994 Dec 2;127(6):2021–2036. doi: 10.1083/jcb.127.6.2021

TGF-beta induced transdifferentiation of mammary epithelial cells to mesenchymal cells: involvement of type I receptors

PMCID: PMC2120317  PMID: 7806579

Abstract

The secreted polypeptide transforming growth factor-beta (TGF-beta) exerts its multiple activities through type I and II cell surface receptors. In epithelial cells, activation of the TGF-beta signal transduction pathways leads to inhibition of cell proliferation and an increase in extracellular matrix production. TGF-beta is widely expressed during development and its biological activity has been implicated in epithelial-mesenchymal interactions, e.g., in branching morphogenesis of the lung, kidney, and mammary gland, and in inductive events between mammary epithelium and stroma. In the present study, we investigated the effects of TGF-beta on mouse mammary epithelial cells in vitro. TGF-beta reversibly induced an alteration in the differentiation of normal mammary epithelial NMuMG cells from epithelial to fibroblastic phenotype. The change in cell morphology correlated with (a) decreased expression of the epithelial markers E- cadherin, ZO-1, and desmoplakin I and II; (b) increased expression of mesenchymal markers, such as fibronectin; and (c) a fibroblast-like reorganization of actin fibers. This phenotypic differentiation displays the hallmarks of an epithelial to mesenchymal transdifferentiation event. Since NMuMG cells make high levels of the type I TGF-beta receptor Tsk7L, yet lack expression of the ALK-5/R4 type I receptor which has been reported to mediate TGF-beta responsiveness, we evaluated the role of the Tsk7L receptor in TGF-beta- mediated transdifferentiation. We generated NMuMG cells that stably overexpress a truncated Tsk7L type I receptor that lacks most of the cytoplasmic kinase domain, thus function as a dominant negative mutant. These transfected cells no longer underwent epithelial to mesenchymal morphological change upon exposure to TGF-beta, yet still displayed some TGF-beta-mediated responses. We conclude that TGF-beta has the ability to modulate E-cadherin expression and induce a reversible epithelial to mesenchymal transdifferentiation in epithelial cells. Unlike other transdifferentiating growth factors, such as bFGF and HGF, these changes are accompanied by growth inhibition. Our results also implicate the Tsk7L type I receptor as mediating the TGF-beta-induced epithelial to mesenchymal transition.

Full Text

The Full Text of this article is available as a PDF (14.9 MB).

Selected References

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

  1. Attisano L., Cárcamo J., Ventura F., Weis F. M., Massagué J., Wrana J. L. Identification of human activin and TGF beta type I receptors that form heteromeric kinase complexes with type II receptors. Cell. 1993 Nov 19;75(4):671–680. doi: 10.1016/0092-8674(93)90488-c. [DOI] [PubMed] [Google Scholar]
  2. Bachem M. G., Sell K. M., Melchior R., Kropf J., Eller T., Gressner A. M. Tumor necrosis factor alpha (TNF alpha) and transforming growth factor beta 1 (TGF beta 1) stimulate fibronectin synthesis and the transdifferentiation of fat-storing cells in the rat liver into myofibroblasts. Virchows Arch B Cell Pathol Incl Mol Pathol. 1993;63(2):123–130. doi: 10.1007/BF02899251. [DOI] [PubMed] [Google Scholar]
  3. Bassing C. H., Howe D. J., Segarini P. R., Donahoe P. K., Wang X. F. A single heteromeric receptor complex is sufficient to mediate biological effects of transforming growth factor-beta ligands. J Biol Chem. 1994 May 27;269(21):14861–14864. [PubMed] [Google Scholar]
  4. Bassing C. H., Yingling J. M., Howe D. J., Wang T., He W. W., Gustafson M. L., Shah P., Donahoe P. K., Wang X. F. A transforming growth factor beta type I receptor that signals to activate gene expression. Science. 1994 Jan 7;263(5143):87–89. doi: 10.1126/science.8272871. [DOI] [PubMed] [Google Scholar]
  5. Behrens J., Mareel M. M., Van Roy F. M., Birchmeier W. Dissecting tumor cell invasion: epithelial cells acquire invasive properties after the loss of uvomorulin-mediated cell-cell adhesion. J Cell Biol. 1989 Jun;108(6):2435–2447. doi: 10.1083/jcb.108.6.2435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brand T., MacLellan W. R., Schneider M. D. A dominant-negative receptor for type beta transforming growth factors created by deletion of the kinase domain. J Biol Chem. 1993 Jun 5;268(16):11500–11503. [PubMed] [Google Scholar]
  7. Burdsal C. A., Damsky C. H., Pedersen R. A. The role of E-cadherin and integrins in mesoderm differentiation and migration at the mammalian primitive streak. Development. 1993 Jul;118(3):829–844. doi: 10.1242/dev.118.3.829. [DOI] [PubMed] [Google Scholar]
  8. Chen R. H., Ebner R., Derynck R. Inactivation of the type II receptor reveals two receptor pathways for the diverse TGF-beta activities. Science. 1993 May 28;260(5112):1335–1338. doi: 10.1126/science.8388126. [DOI] [PubMed] [Google Scholar]
  9. Cárcamo J., Weis F. M., Ventura F., Wieser R., Wrana J. L., Attisano L., Massagué J. Type I receptors specify growth-inhibitory and transcriptional responses to transforming growth factor beta and activin. Mol Cell Biol. 1994 Jun;14(6):3810–3821. doi: 10.1128/mcb.14.6.3810. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Daniel C. W., Silberstein G. B., Van Horn K., Strickland P., Robinson S. TGF-beta 1-induced inhibition of mouse mammary ductal growth: developmental specificity and characterization. Dev Biol. 1989 Sep;135(1):20–30. doi: 10.1016/0012-1606(89)90154-1. [DOI] [PubMed] [Google Scholar]
  11. Ebner R., Chen R. H., Lawler S., Zioncheck T., Derynck R. Determination of type I receptor specificity by the type II receptors for TGF-beta or activin. Science. 1993 Nov 5;262(5135):900–902. doi: 10.1126/science.8235612. [DOI] [PubMed] [Google Scholar]
  12. Ebner R., Chen R. H., Shum L., Lawler S., Zioncheck T. F., Lee A., Lopez A. R., Derynck R. Cloning of a type I TGF-beta receptor and its effect on TGF-beta binding to the type II receptor. Science. 1993 May 28;260(5112):1344–1348. doi: 10.1126/science.8388127. [DOI] [PubMed] [Google Scholar]
  13. Edwards D. R., Murphy G., Reynolds J. J., Whitham S. E., Docherty A. J., Angel P., Heath J. K. Transforming growth factor beta modulates the expression of collagenase and metalloproteinase inhibitor. EMBO J. 1987 Jul;6(7):1899–1904. doi: 10.1002/j.1460-2075.1987.tb02449.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Eguchi G., Kodama R. Transdifferentiation. Curr Opin Cell Biol. 1993 Dec;5(6):1023–1028. doi: 10.1016/0955-0674(93)90087-7. [DOI] [PubMed] [Google Scholar]
  15. Eguchi G., Okada T. S. Differentiation of lens tissue from the progeny of chick retinal pigment cells cultured in vitro: a demonstration of a switch of cell types in clonal cell culture. Proc Natl Acad Sci U S A. 1973 May;70(5):1495–1499. doi: 10.1073/pnas.70.5.1495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Engler-Blum G., Meier M., Frank J., Müller G. A. Reduction of background problems in nonradioactive northern and Southern blot analyses enables higher sensitivity than 32P-based hybridizations. Anal Biochem. 1993 May 1;210(2):235–244. doi: 10.1006/abio.1993.1189. [DOI] [PubMed] [Google Scholar]
  17. Fafeur V., O'Hara B., Böhlen P. A glycosylation-deficient endothelial cell mutant with modified responses to transforming growth factor-beta and other growth inhibitory cytokines: evidence for multiple growth inhibitory signal transduction pathways. Mol Biol Cell. 1993 Feb;4(2):135–144. doi: 10.1091/mbc.4.2.135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Franzén P., ten Dijke P., Ichijo H., Yamashita H., Schulz P., Heldin C. H., Miyazono K. Cloning of a TGF beta type I receptor that forms a heteromeric complex with the TGF beta type II receptor. Cell. 1993 Nov 19;75(4):681–692. doi: 10.1016/0092-8674(93)90489-d. [DOI] [PubMed] [Google Scholar]
  19. Frixen U. H., Behrens J., Sachs M., Eberle G., Voss B., Warda A., Löchner D., Birchmeier W. E-cadherin-mediated cell-cell adhesion prevents invasiveness of human carcinoma cells. J Cell Biol. 1991 Apr;113(1):173–185. doi: 10.1083/jcb.113.1.173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Gavrilović J., Moens G., Thiery J. P., Jouanneau J. Expression of transfected transforming growth factor alpha induces a motile fibroblast-like phenotype with extracellular matrix-degrading potential in a rat bladder carcinoma cell line. Cell Regul. 1990 Dec;1(13):1003–1014. doi: 10.1091/mbc.1.13.1003. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Geiser A. G., Burmester J. K., Webbink R., Roberts A. B., Sporn M. B. Inhibition of growth by transforming growth factor-beta following fusion of two nonresponsive human carcinoma cell lines. Implication of the type II receptor in growth inhibitory responses. J Biol Chem. 1992 Feb 5;267(4):2588–2593. [PubMed] [Google Scholar]
  22. Greenburg G., Hay E. D. Cytodifferentiation and tissue phenotype change during transformation of embryonic lens epithelium to mesenchyme-like cells in vitro. Dev Biol. 1986 Jun;115(2):363–379. doi: 10.1016/0012-1606(86)90256-3. [DOI] [PubMed] [Google Scholar]
  23. Greenburg G., Hay E. D. Cytoskeleton and thyroglobulin expression change during transformation of thyroid epithelium to mesenchyme-like cells. Development. 1988 Mar;102(3):605–622. doi: 10.1242/dev.102.3.605. [DOI] [PubMed] [Google Scholar]
  24. He W. W., Gustafson M. L., Hirobe S., Donahoe P. K. Developmental expression of four novel serine/threonine kinase receptors homologous to the activin/transforming growth factor-beta type II receptor family. Dev Dyn. 1993 Feb;196(2):133–142. doi: 10.1002/aja.1001960207. [DOI] [PubMed] [Google Scholar]
  25. Heine U. I., Munoz E. F., Flanders K. C., Roberts A. B., Sporn M. B. Colocalization of TGF-beta 1 and collagen I and III, fibronectin and glycosaminoglycans during lung branching morphogenesis. Development. 1990 May;109(1):29–36. doi: 10.1242/dev.109.1.29. [DOI] [PubMed] [Google Scholar]
  26. Heine U., Munoz E. F., Flanders K. C., Ellingsworth L. R., Lam H. Y., Thompson N. L., Roberts A. B., Sporn M. B. Role of transforming growth factor-beta in the development of the mouse embryo. J Cell Biol. 1987 Dec;105(6 Pt 2):2861–2876. doi: 10.1083/jcb.105.6.2861. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. Hosobuchi M., Stampfer M. R. Effects of transforming growth factor beta on growth of human mammary epithelial cells in culture. In Vitro Cell Dev Biol. 1989 Aug;25(8):705–713. doi: 10.1007/BF02623723. [DOI] [PubMed] [Google Scholar]
  29. Ignotz R. A., Heino J., Massagué J. Regulation of cell adhesion receptors by transforming growth factor-beta. Regulation of vitronectin receptor and LFA-1. J Biol Chem. 1989 Jan 5;264(1):389–392. [PubMed] [Google Scholar]
  30. Kingsley D. M. The TGF-beta superfamily: new members, new receptors, and new genetic tests of function in different organisms. Genes Dev. 1994 Jan;8(2):133–146. doi: 10.1101/gad.8.2.133. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Laiho M., Weis M. B., Massagué J. Concomitant loss of transforming growth factor (TGF)-beta receptor types I and II in TGF-beta-resistant cell mutants implicates both receptor types in signal transduction. J Biol Chem. 1990 Oct 25;265(30):18518–18524. [PubMed] [Google Scholar]
  33. Lawler S., Candia A. F., Ebner R., Shum L., Lopez A. R., Moses H. L., Wright C. V., Derynck R. The murine type II TGF-beta receptor has a coincident embryonic expression and binding preference for TGF-beta 1. Development. 1994 Jan;120(1):165–175. doi: 10.1242/dev.120.1.165. [DOI] [PubMed] [Google Scholar]
  34. Lin H. Y., Wang X. F., Ng-Eaton E., Weinberg R. A., Lodish H. F. Expression cloning of the TGF-beta type II receptor, a functional transmembrane serine/threonine kinase. Cell. 1992 Feb 21;68(4):775–785. doi: 10.1016/0092-8674(92)90152-3. [DOI] [PubMed] [Google Scholar]
  35. López-Casillas F., Cheifetz S., Doody J., Andres J. L., Lane W. S., Massagué J. Structure and expression of the membrane proteoglycan betaglycan, a component of the TGF-beta receptor system. Cell. 1991 Nov 15;67(4):785–795. doi: 10.1016/0092-8674(91)90073-8. [DOI] [PubMed] [Google Scholar]
  36. López-Casillas F., Wrana J. L., Massagué J. Betaglycan presents ligand to the TGF beta signaling receptor. Cell. 1993 Jul 2;73(7):1435–1444. doi: 10.1016/0092-8674(93)90368-z. [DOI] [PubMed] [Google Scholar]
  37. Matsuzaki F., Mège R. M., Jaffe S. H., Friedlander D. R., Gallin W. J., Goldberg J. I., Cunningham B. A., Edelman G. M. cDNAs of cell adhesion molecules of different specificity induce changes in cell shape and border formation in cultured S180 cells. J Cell Biol. 1990 Apr;110(4):1239–1252. doi: 10.1083/jcb.110.4.1239. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Moustakas A., Lin H. Y., Henis Y. I., Plamondon J., O'Connor-McCourt M. D., Lodish H. F. The transforming growth factor beta receptors types I, II, and III form hetero-oligomeric complexes in the presence of ligand. J Biol Chem. 1993 Oct 25;268(30):22215–22218. [PubMed] [Google Scholar]
  39. Nagafuchi A., Shirayoshi Y., Okazaki K., Yasuda K., Takeichi M. Transformation of cell adhesion properties by exogenously introduced E-cadherin cDNA. Nature. 1987 Sep 24;329(6137):341–343. doi: 10.1038/329341a0. [DOI] [PubMed] [Google Scholar]
  40. Ohtsuki M., Massagué J. Evidence for the involvement of protein kinase activity in transforming growth factor-beta signal transduction. Mol Cell Biol. 1992 Jan;12(1):261–265. doi: 10.1128/mcb.12.1.261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Pierce D. F., Jr, Johnson M. D., Matsui Y., Robinson S. D., Gold L. I., Purchio A. F., Daniel C. W., Hogan B. L., Moses H. L. Inhibition of mammary duct development but not alveolar outgrowth during pregnancy in transgenic mice expressing active TGF-beta 1. Genes Dev. 1993 Dec;7(12A):2308–2317. doi: 10.1101/gad.7.12a.2308. [DOI] [PubMed] [Google Scholar]
  42. Potts J. D., Runyan R. B. Epithelial-mesenchymal cell transformation in the embryonic heart can be mediated, in part, by transforming growth factor beta. Dev Biol. 1989 Aug;134(2):392–401. doi: 10.1016/0012-1606(89)90111-5. [DOI] [PubMed] [Google Scholar]
  43. Roberts C. J., Birkenmeier T. M., McQuillan J. J., Akiyama S. K., Yamada S. S., Chen W. T., Yamada K. M., McDonald J. A. Transforming growth factor beta stimulates the expression of fibronectin and of both subunits of the human fibronectin receptor by cultured human lung fibroblasts. J Biol Chem. 1988 Apr 5;263(10):4586–4592. [PubMed] [Google Scholar]
  44. Robertson P. L., Markovac J., Datta S. C., Goldstein G. W. Transforming growth factor beta stimulates phosphoinositol metabolism and translocation of protein kinase C in cultured astrocytes. Neurosci Lett. 1988 Oct 31;93(1):107–113. doi: 10.1016/0304-3940(88)90021-3. [DOI] [PubMed] [Google Scholar]
  45. Robinson S. D., Silberstein G. B., Roberts A. B., Flanders K. C., Daniel C. W. Regulated expression and growth inhibitory effects of transforming growth factor-beta isoforms in mouse mammary gland development. Development. 1991 Nov;113(3):867–878. doi: 10.1242/dev.113.3.867. [DOI] [PubMed] [Google Scholar]
  46. Rogers S. A., Ryan G., Purchio A. F., Hammerman M. R. Metanephric transforming growth factor-beta 1 regulates nephrogenesis in vitro. Am J Physiol. 1993 Jun;264(6 Pt 2):F996–1002. doi: 10.1152/ajprenal.1993.264.6.F996. [DOI] [PubMed] [Google Scholar]
  47. Schwarzbauer J. E., Tamkun J. W., Lemischka I. R., Hynes R. O. Three different fibronectin mRNAs arise by alternative splicing within the coding region. Cell. 1983 Dec;35(2 Pt 1):421–431. doi: 10.1016/0092-8674(83)90175-7. [DOI] [PubMed] [Google Scholar]
  48. Silberstein G. B., Daniel C. W. Reversible inhibition of mammary gland growth by transforming growth factor-beta. Science. 1987 Jul 17;237(4812):291–293. doi: 10.1126/science.3474783. [DOI] [PubMed] [Google Scholar]
  49. Silberstein G. B., Strickland P., Coleman S., Daniel C. W. Epithelium-dependent extracellular matrix synthesis in transforming growth factor-beta 1-growth-inhibited mouse mammary gland. J Cell Biol. 1990 Jun;110(6):2209–2219. doi: 10.1083/jcb.110.6.2209. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Somers C. E., Mosher D. F. Protein kinase C modulation of fibronectin matrix assembly. J Biol Chem. 1993 Oct 25;268(30):22277–22280. [PubMed] [Google Scholar]
  51. Sommers C. L., Heckford S. E., Skerker J. M., Worland P., Torri J. A., Thompson E. W., Byers S. W., Gelmann E. P. Loss of epithelial markers and acquisition of vimentin expression in adriamycin- and vinblastine-resistant human breast cancer cell lines. Cancer Res. 1992 Oct 1;52(19):5190–5197. [PubMed] [Google Scholar]
  52. Stampfer M. R., Yaswen P., Alhadeff M., Hosoda J. TGF beta induction of extracellular matrix associated proteins in normal and transformed human mammary epithelial cells in culture is independent of growth effects. J Cell Physiol. 1993 Apr;155(1):210–221. doi: 10.1002/jcp.1041550127. [DOI] [PubMed] [Google Scholar]
  53. Tomoda T., Kudoh T., Noma T., Nakazawa A., Muramatsu M., Arai K. Molecular cloning of a mouse counterpart for human TGF-beta type I receptor. Biochem Biophys Res Commun. 1994 Feb 15;198(3):1054–1062. doi: 10.1006/bbrc.1994.1150. [DOI] [PubMed] [Google Scholar]
  54. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Tsarfaty I., Rong S., Resau J. H., Rulong S., da Silva P. P., Vande Woude G. F. The Met proto-oncogene mesenchymal to epithelial cell conversion. Science. 1994 Jan 7;263(5143):98–101. doi: 10.1126/science.7505952. [DOI] [PubMed] [Google Scholar]
  56. Vallés A. M., Tucker G. C., Thiery J. P., Boyer B. Alternative patterns of mitogenesis and cell scattering induced by acidic FGF as a function of cell density in a rat bladder carcinoma cell line. Cell Regul. 1990 Dec;1(13):975–988. doi: 10.1091/mbc.1.13.975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Wang X. F., Lin H. Y., Ng-Eaton E., Downward J., Lodish H. F., Weinberg R. A. Expression cloning and characterization of the TGF-beta type III receptor. Cell. 1991 Nov 15;67(4):797–805. doi: 10.1016/0092-8674(91)90074-9. [DOI] [PubMed] [Google Scholar]
  58. Wieser R., Attisano L., Wrana J. L., Massagué J. Signaling activity of transforming growth factor beta type II receptors lacking specific domains in the cytoplasmic region. Mol Cell Biol. 1993 Dec;13(12):7239–7247. doi: 10.1128/mcb.13.12.7239. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Wrana J. L., Attisano L., Cárcamo J., Zentella A., Doody J., Laiho M., Wang X. F., Massagué J. TGF beta signals through a heteromeric protein kinase receptor complex. Cell. 1992 Dec 11;71(6):1003–1014. doi: 10.1016/0092-8674(92)90395-s. [DOI] [PubMed] [Google Scholar]
  60. Zentella A., Weis F. M., Ralph D. A., Laiho M., Massagué J. Early gene responses to transforming growth factor-beta in cells lacking growth-suppressive RB function. Mol Cell Biol. 1991 Oct;11(10):4952–4958. doi: 10.1128/mcb.11.10.4952. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. ten Dijke P., Yamashita H., Ichijo H., Franzén P., Laiho M., Miyazono K., Heldin C. H. Characterization of type I receptors for transforming growth factor-beta and activin. Science. 1994 Apr 1;264(5155):101–104. doi: 10.1126/science.8140412. [DOI] [PubMed] [Google Scholar]

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

RESOURCES