Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Short Report
  • Published:

TGF-β1 acts as a tumor suppressor of human malignant keratinocytes independently of Smad 4 expression and ligand-induced G1 arrest

Oncogene volume 21, pages 1616–1624 (2002)Cite this article

Abstract

This study examined the role of TGF-β1 in human keratinocyte malignancy. Two carcinoma-derived human oral keratinocyte cell lines, BICR 31 and H314, were selected on the basis of their known resistance to TGF-β1-induced G1 arrest, the presence of wild type TGF-β cell surface receptors and normal Ras. Smad 4 protein was undetectable in both cell lines, but Smad 2 and Smad 3 were expressed at levels comparable with a fully TGF-β responsive cell line, and treatment of the cells with TGF-β1 resulted in the phosphorylation of Smad 2. Treatment with exogenous TGF-β1 resulted in a failure to induce transcription from an artificial Smad-dependent promoter and a failure to down-regulate c-myc, but resulted in an up-regulation of AP-1 associated genes (Fra-1, JunB and fibronectin). Transient transfection of Smad 4 into BICR 31 restored TGF-β1-induced growth inhibition and Smad-dependent transcriptional activation. Protracted treatment of cells with exogenous TGF-β1 resulted in the attenuation of cell growth in vitro. To over-express TGF-β1, both cell lines were transfected with latent TGF-β1 cDNA; neutralization studies of conditioned media demonstrated that whilst the majority of the peptide was in the latent form, a small proportion was present as the active peptide. Cells that over-expressed endogenous TGF-β1 grew more slowly in vitro compared to both the vector-only controls and cells that did not over-express the peptide. Orthotopic transplantation of cells that over-expressed endogenous TGF-β1 to the floor of the mouth in athymic mice resulted in marked inhibition of primary tumor formation compared to controls. Expression of a dominant-negative TGF-β type II receptor in cells that over-expressed endogenous TGF-β1 resulted in enhanced cell growth in vitro and diminished the tumor suppressor effect of the ligand in vivo, indicating that the endogenous TGF-β1 was acting in an autocrine capacity. The results demonstrate that over-expression of endogenous TGF-β1 in human malignant oral keratinocytes leads to growth inhibition in vivo and tumor suppression in vitro by mechanisms that are independent of Smad 4 expression and TGF-β1-induced G1 arrest.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3

Similar content being viewed by others

References

  • Amendt C, Schirmacher P, Weber H, Blessing M . 1998 Oncogene 17: 25–34

  • Burns JE, Baird MC, Clark LJ, Burns PA, Edington K, Chapman C, Mitchell R, Robertson G, Soutar D, Parkinson EK . 1993 Br. J. Cancer 67: 1274–1284

  • Chen CR, Kang Y, Massague J . 2001a Proc. Natl. Acad. Sci. USA 98: 992–999

  • Chen T, Yan W, Wells RG, Rimm DL, McNiff J, Leffell D, Reiss M . 2001b Int. J. Cancer 93: 653–661

  • Clark LJ, Edington K, Swan IR, McLay KA, Newlands WJ, Wills LC, Young HA, Johnstone PW, Mitchel R, Robertson G, Soutar D, Parkinson EK, Birnie GD . 1993 Br. J. Cancer 68: 617–620

  • Cui W, Kemp CJ, Duffie E, Balmain A, Ackhurst RJ . 1994 Cancer Res. 54: 5831–5836

  • Davies M, Prime SS, Stone AM, Heung Y-L, Huntley SP, Matthews JB, Eveson JW, Paterson IC . 1997 Int. J. Cancer 73: 68–74

  • De Caestecker MP, Piek E, Roberts AB . 2000 J. Natl. Cancer Inst. 92: 1388–1402

  • Derynck R, Zhang Y, Feng XH . 1998 Cell 95: 737–740

  • De Winter JP, Roelen BAJ, ten Dijke P, van der Burg B, van den Eijnden-van Raaij AJM . 1997 Oncogene 14: 1891–1899

  • Fink SP, Swinler SE, Lutterbaugh JD, Massague J, Thiagalingam S, Kinzler KW, Vogelstein B, Willson JKV, Markowitz S . 2001 Cancer Res. 61: 256–260

  • Garrigue-Antar L, Munoz-Antonia T, Antonia SJ, Gesmonde J, Vellucci VF, Reiss M . 1995 Cancer Res. 55: 3982–3987

  • Glick AB, Kulkarni AB, Tennenbaum T, Hennings H, Flanders KC, O'Reilly M, Sporn MB, Karlsson S, Yuspa SH . 1993 Proc. Natl. Acad. Sci. USA 90: 6076–6080

  • Glick AB, Lee MM, Darwiche N, Kulkarni AB, Karlsson S, Yuspa SH . 1994 Genes Dev. 15: 2429–2440

  • Hahn SA, Schutte M, Hoque AT, Moskaluk CA, da Costa LT, Rozenblum E, Weinstein CL, Fischer A, Yeo CJ, Hruban RH, Kern SE . 1996 Science 271: 350–353

  • Hocevar BA, Brown TL, Howe PH . 1999 EMBO J. 18: 1345–1356

  • Howell M, Itoh F, Pierreux CE, Valgeirsdottir S, Itoh S, ten Dijke P, Hill CS . 1999 Dev. Biol. 214: 354–369

  • Jonk LJ, Itoh S, Heldin CH, ten Dijke P, Kruijer W . 1998 J. Biol. Chem. 273: 21145–21152

  • Kim SK, Fan Y, Papadimitrikopoulou V, Clayman G, Hittelman WN, Hong WK, Lotan R, Mao L . 1996 Cancer Res. 56: 2519–2521

  • Kinzler KW, Vogelstein B . 1997 Nature 386: 761–763

  • Korchynskyi O, Landstrom M, Stoika R, Funa K, Heldin CH, ten Dijke P, Souchelnytskyi S . 1999 Int. J. Cancer 82: 197–202

  • Le Dai J, Schutte M, Bansal RK, Wilentz RE, Sugar AY, Kern SE . 1999 Mol. Carcinog. 26: 37–43

  • Malliri A, Yeudall WA, Nikolic M, Crouch DH, Parkinson E, Ozanne B . 1996 Cell Growth Differ. 7: 1291–1304

  • Masuyama N, Hanafusa H, Kusakabe M, Shibuya H, Nishida E . 1999 J. Biol. Chem. 274: 12163–12170

  • Paterson IC, Matthews JB, Huntley S, Robinson CM, Fahey M, Parkinson EK, Prime SS . 2001 J. Pathol. 193: 458–467

  • Paterson IC, Patel V, Sandy JR, Prime SS, Yeudall WA . 1995 Br. J. Cancer 72: 922–927

  • Petritsch C, Beug H, Balmain A, Oft M . 2000 Genes Dev. 14: 3093–3101

  • Pierreux CE, Nicolas FJ, Hill CS . 2000 Mol. Cell. Biol. 20: 9041–9054

  • Prime SS, Matthews JB, Patel V, Game SM, Donnelly MJ, Stone A, Paterson IC, Sandy JR, Yeudall WA . 1994 Int. J. Cancer 56: 406–412

  • Qing J, Zhang Y, Derynck R . 2000 J. Biol. Chem. 275: 38802–38812

  • Riggins GJ, Kinzler KW, Vogelstein B, Thiagalingam S . 1997 Cancer Res. 57: 2578–2580

  • Schutte M, Hruban RH, Hedrick L, Cho KR, Nadasdy GM, Weinstein CL, Bova GS, Isaacs WB, Cairns P, Nawroz H, Sidransky D, Casero Jr RA, Meltzer PS, Hahn SA, Kern SE . 1996 Cancer Res. 56: 2527–2530

  • Sirard C, Kim S, Mirtsos C, Tadich P, Hoodless PA, Itie A, Maxson R, Wrana JL, Mak TW . 2000 J. Biol. Chem. 275: 2063–2070

  • Takaku K, Oshima M, Miyoshi H, Matsui M, Seldin MF, Taketo MM . 1999 Cell 92: 645–656

  • Wang D, Song H, Evans JA, Lang JC, Schuller DE, Weghorst CM . 1997 Carcinogenesis 18: 2285–2290

  • Wilentz RE, Su GH, Dai JL, Sparks AB, Argani P, Sohn TA, Yeo CJ, Kern SE, Hruban RH . 2000a Am. J. Pathol. 156: 37–43

  • Wilentz RE, Iacobuzio-Donahue CA, Argani P, McCarthy DM, Parsons JL, Yoe CJ, Kern SE, Hruban RH . 2000b Cancer Res. 60: 2002–2006

  • Wong C, Rougier-Chapman EM, Frederick JP, Datto MB, Liberati NT, Li JM, Wang XF . 1999 Cell Biol. Mol. 19: 1821–1830

  • Xu X, Brodie SG, Yang X, Im YH, Parks WT, Chen L, Zhou YX, Weinstein M, Kim SJ, Deng CX . 2000 Oncogene 19: 1868–1874

  • Yamamura Y, Hua X, Bergelson S, Lodish HF . 2000 J. Biol. Chem. 275: 36295–36302

  • Yan W, Vellucci VF, Reiss M . 2000 Oncol. Res. 12: 157–167

  • Yeudall WA, Torrance LK, Elsegood KA, Speight P, Scully C, Prime SS . 1993 Oral Oncol. Eur. J. Cancer 29B: 63–67

  • Yeudall WA, Paterson IC, Patel V, Prime SS . 1995 Oral. Oncol. Eur. J. Cancer 31B: 136–143

  • Zawel L, Dai JL, Buckhaults P, Zhou S, Kinzler KW, Vogelstein B, Kern SE . 1998 Mol. Cell. 1: 611–617

  • Zhang Y, Feng XH, Derynck R . 1998 Nature 394: 909–913

Download references

Acknowledgements

We thank Prof B Vogelstein for the pSBC.Luc and the pMBE.Luc reporter plasmids and Prof J Massague for the Smad 4 expression construct. The pCM41c-myc, pBR18S, pSKIIfra1, pBR322αactin and fibronectin plasmids were kind gifts from Dr A Ganderillas (ICRF, UK).

Author information

Author notes

  1. Ian C Paterson and Maria Davies: Equal contribution by these authors.

Authors and Affiliations

  1. Department of Oral and Dental Science, University of Bristol, Bristol, BS1 2LY, UK

    Ian C Paterson, Maria Davies, Andrea Stone, Suzy Huntley, Emily Smith, Miranda Pring, John W Eveson, C Max Robinson & Stephen S Prime

  2. CRC Beatson Institute for Cancer Research, Glasgow, G61 1BD, UK

    E Kenneth Parkinson

Authors
  1. Ian C Paterson
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Maria Davies
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Andrea Stone
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Suzy Huntley
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Emily Smith
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Miranda Pring
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. John W Eveson
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. C Max Robinson
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. E Kenneth Parkinson
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Stephen S Prime
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ian C Paterson.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Paterson, I., Davies, M., Stone, A. et al. TGF-β1 acts as a tumor suppressor of human malignant keratinocytes independently of Smad 4 expression and ligand-induced G1 arrest. Oncogene 21, 1616–1624 (2002). https://doi.org/10.1038/sj.onc.1205217

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1205217

Keywords

Search

Advanced search

Quick links