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.

  • Original Article
  • Published:

TGF-beta receptor type-2 expression in cancer-associated fibroblasts regulates breast cancer cell growth and survival and is a prognostic marker in pre-menopausal breast cancer

Abstract

Transforming growth factor-beta (TGF-β) is a pleiotropic cytokine with the capability to act as tumour suppressor or tumour promoter depending on the cellular context. TGF-beta receptor type-2 (TGFBR2) is the ligand-binding receptor for all members of the TGF-β family. Data from mouse model experiments demonstrated that loss of Tgfbr2 expression in mammary fibroblasts was linked to tumour initiation and metastasis. Using a randomised tamoxifen trial cohort including in total 564 invasive breast carcinomas, we examined TGFBR2 expression (n=252) and phosphorylation level of downstream target SMAD2 (pSMAD2) (n=319) in cancer-associated fibroblasts (CAFs) and assessed links to clinicopathological markers, prognostic and treatment-predictive values. The study revealed that CAF-specific TGFBR2 expression correlated with improved recurrence-free survival. Multivariate analysis confirmed CAF-TGFBR2 to be an independent prognostic marker (multivariate Cox regression, hazard ratio: 0.534, 95% (CI): 0.360–0.793, P=0.002). CAF-specific pSMAD2 levels, however, did not associate with survival outcome. Experimentally, TGF-β signalling in fibroblasts was modulated using a TGF-β ligand and inhibitor or through lentiviral short hairpin RNA-mediated TGFBR2-specific knockdown. To determine the role of fibroblastic TGF-β pathway on breast cancer cells, we used cell contact-dependent cell growth and clonogenicity assays, which showed that knockdown of TGFBR2 in CAFs resulted in increased cell growth, proliferation and clonogenic survival. Further, in a mouse model transfected CAFs were co-injected with MCF7 and tumour weight and proportion was monitored. We found that mouse xenograft tumours comprising TGFBR2 knockdown fibroblasts were slightly bigger and displayed increased tumour cell capacity. Overall, our data demonstrate that fibroblast-related biomarkers possess clinically relevant information and that fibroblasts confer effects on breast cancer cell growth and survival. Regulation of tumour–stromal cross-talk through fibroblastic TGF-β pathway may depend on fibroblast phenotype, emphasising the importance to characterise tumour microenvironment subtypes.

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

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

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

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5

Similar content being viewed by others

References

  1. Bierie B, Moses HL . Tumour microenvironment: TGFbeta: the molecular Jekyll and Hyde of cancer. Nat Rev Cancer 2006; 6: 506–520.

    Article  CAS  PubMed  Google Scholar 

  2. Siegel PM, Massague J . Cytostatic and apoptotic actions of TGF-beta in homeostasis and cancer. Nat Rev Cancer 2003; 3: 807–821.

    Article  CAS  PubMed  Google Scholar 

  3. Massague J . TGFbeta in Cancer. Cell 2008; 134: 215–230.

    CAS  PubMed  PubMed Central  Google Scholar 

  4. Derynck R, Zhang YE . Smad-dependent and Smad-independent pathways in TGF-beta family signalling. Nature 2003; 425: 577–584.

    Article  CAS  PubMed  Google Scholar 

  5. De Wever O, Demetter P, Mareel M, Bracke M . Stromal myofibroblasts are drivers of invasive cancer growth. Int J Cancer 2008; 123: 2229–2238.

    Article  CAS  PubMed  Google Scholar 

  6. De Wever O, Mareel M . Role of tissue stroma in cancer cell invasion. J Pathol 2003; 200: 429–447.

    Article  CAS  PubMed  Google Scholar 

  7. Bhowmick NA, Neilson EG, Moses HL . Stromal fibroblasts in cancer initiation and progression. Nature 2004; 432: 332–337.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Mueller MM, Fusenig NE . Friends or foes—bipolar effects of the tumour stroma in cancer. Nat Rev Cancer 2004; 4: 839–849.

    Article  CAS  PubMed  Google Scholar 

  9. Bhowmick NA, Chytil A, Plieth D, Gorska AE, Dumont N, Shappell S et al. TGF-beta signaling in fibroblasts modulates the oncogenic potential of adjacent epithelia. Science 2004; 303: 848–851.

    Article  CAS  PubMed  Google Scholar 

  10. Cheng N, Bhowmick NA, Chytil A, Gorksa AE, Brown KA, Muraoka R et al. Loss of TGF-beta type II receptor in fibroblasts promotes mammary carcinoma growth and invasion through upregulation of TGF-alpha-, MSP- and HGF-mediated signaling networks. Oncogene 2005; 24: 5053–5068.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Cheng N, Chytil A, Shyr Y, Joly A, Moses HL . Enhanced hepatocyte growth factor signaling by type II transforming growth factor-beta receptor knockout fibroblasts promotes mammary tumorigenesis. Cancer Res 2007; 67: 4869–4877.

    Article  CAS  PubMed  Google Scholar 

  12. Cheng N, Chytil A, Shyr Y, Joly A, Moses HL . Transforming growth factor-beta signaling-deficient fibroblasts enhance hepatocyte growth factor signaling in mammary carcinoma cells to promote scattering and invasion. Mol Cancer Res 2008; 6: 1521–1533.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Bacman D, Merkel S, Croner R, Papadopoulos T, Brueckl W, Dimmler A . TGF-beta receptor 2 downregulation in tumour-associated stroma worsens prognosis and high-grade tumours show more tumour-associated macrophages and lower TGF-beta1 expression in colon carcinoma: a retrospective study. BMC Cancer 2007; 7: 156.

    Article  PubMed  PubMed Central  Google Scholar 

  14. Casbas-Hernandez P, Fleming JM, Troester MA . Gene expression analysis of in vitro cocultures to study interactions between breast epithelium and stroma. J Biomed Biotechnol 2011; 2011: 520987.

    Article  PubMed  PubMed Central  Google Scholar 

  15. Stuelten CH, Busch JI, Tang B, Flanders KC, Oshima A, Sutton E et al. Transient tumor-fibroblast interactions increase tumor cell malignancy by a TGF-Beta mediated mechanism in a mouse xenograft model of breast cancer. PLoS One 2010; 5: e9832.

    Article  PubMed  PubMed Central  Google Scholar 

  16. Hanahan D, Weinberg RA . The hallmarks of cancer. Cell 2000; 100: 57–70.

    Article  CAS  PubMed  Google Scholar 

  17. Neve RM, Chin K, Fridlyand J, Yeh J, Baehner FL, Fevr T et al. A collection of breast cancer cell lines for the study of functionally distinct cancer subtypes. Cancer Cell 2006; 10: 515–527.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Busch S, Ryden L, Stal O, Jirstrom K, Landberg G . Low ERK phosphorylation in cancer-associated fibroblasts is associated with tamoxifen resistance in pre-menopausal breast cancer. PLoS One 2012; 7: e45669.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Desmouliere A, Geinoz A, Gabbiani F, Gabbiani G . Transforming growth factor-beta 1 induces alpha-smooth muscle actin expression in granulation tissue myofibroblasts and in quiescent and growing cultured fibroblasts. J Cell Biol 1993; 122: 103–111.

    Article  CAS  PubMed  Google Scholar 

  20. Kojima Y, Acar A, Eaton EN, Mellody KT, Scheel C, Ben-Porath I et al. Autocrine TGF-beta and stromal cell-derived factor-1 (SDF-1) signaling drives the evolution of tumor-promoting mammary stromal myofibroblasts. Proc Natl Acad Sci USA 2010; 107: 20009–20014.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Inman GJ, Nicolas FJ, Callahan JF, Harling JD, Gaster LM, Reith AD et al. SB-431542 is a potent and specific inhibitor of transforming growth factor-beta superfamily type I activin receptor-like kinase (ALK) receptors ALK4, ALK5, and ALK7. Mol Pharmacol 2002; 62: 65–74.

    Article  CAS  PubMed  Google Scholar 

  22. Shi Y, Massague J . Mechanisms of TGF-beta signaling from cell membrane to the nucleus. Cell 2003; 113: 685–700.

    Article  CAS  PubMed  Google Scholar 

  23. Bauer M, Su G, Casper C, He R, Rehrauer W, Friedl A . Heterogeneity of gene expression in stromal fibroblasts of human breast carcinomas and normal breast. Oncogene 2010; 29: 1732–1740.

  24. Sugimoto H, Mundel TM, Kieran MW, Kalluri R . Identification of fibroblast heterogeneity in the tumor microenvironment. Cancer Biol Ther 2006; 5: 1640–1646.

    Article  CAS  PubMed  Google Scholar 

  25. Ryden L, Jonsson PE, Chebil G, Dufmats M, Ferno M, Jirstrom K et al. Two years of adjuvant tamoxifen in premenopausal patients with breast cancer: a randomised, controlled trial with long-term follow-up. Eur J Cancer 2005; 41: 256–264.

    Article  CAS  PubMed  Google Scholar 

  26. Holm C, Rayala S, Jirstrom K, Stal O, Kumar R, Landberg G . Association between Pak1 expression and subcellular localization and tamoxifen resistance in breast cancer patients. J Natl Cancer Inst 2006; 98: 671–680.

    Article  CAS  PubMed  Google Scholar 

  27. Polanska UM, Acar A, Orimo A . Experimental generation of carcinoma-associated fibroblasts (CAFs) from human mammary fibroblasts. J Vis Exp 2011; 56: e3201.

    Google Scholar 

  28. Stewart SA, Dykxhoorn DM, Palliser D, Mizuno H, Yu EY, An DS et al. Lentivirus-delivered stable gene silencing by RNAi in primary cells. RNA 2003; 9: 493–501.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Yamashita S, Takahashi S, McDonell N, Watanabe N, Niwa T, Hosoya K et al. Methylation silencing of transforming growth factor-beta receptor type II in rat prostate cancers. Cancer Res 2008; 68: 2112–2121.

    Article  CAS  PubMed  Google Scholar 

  30. Samoszuk M, Tan J, Chorn G . Clonogenic growth of human breast cancer cells co-cultured in direct contact with serum-activated fibroblasts. Breast Cancer Res 2005; 7: R274–R283.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank Elise Nilsson for excellent technical assistance, the MCRC Biobank for providing us with patient material and Dr Akira Orimo for the kind gift of fibroblast model cell lines and lentiviral plasmids. We would further thank Éamon Hughes for his assistance with the animal work. This study was supported by the Swedish Cancer Society and Breakthrough Breast Cancer UK. The project was further supported by BioCARE—a National Strategic Research Program at University of Gothenburg.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to G Landberg.

Ethics declarations

Competing interests

The authors declare no conflict of interest.

Additional information

Supplementary Information accompanies this paper on the Oncogene website

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Busch, S., Acar, A., Magnusson, Y. et al. TGF-beta receptor type-2 expression in cancer-associated fibroblasts regulates breast cancer cell growth and survival and is a prognostic marker in pre-menopausal breast cancer. Oncogene 34, 27–38 (2015). https://doi.org/10.1038/onc.2013.527

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2013.527

Keywords

This article is cited by

Search

Quick links