Abstract
Tumor growth factor-β (TGF-β) signaling in cancer has been implicated in growth suppression of early lesions and enhancing tumor cell invasion and metastasis. However, the cellular mechanisms that determine this signaling output in individual tumors are still largely unknown. In endothelial cells, TGF-β signaling is modulated by the TGF-β co-receptor endoglin (CD105). Here we demonstrate that endoglin is expressed in a subset of invasive breast cancers and cell lines and is subject to epigenetic silencing by gene methylation. Endoglin downregulation in non-tumorigenic MCF10A breast cells leads to the formation of abnormal acini in 3D culture, but does not promote cell migration or transformation. In contrast, in the presence of activated ErbB2, endoglin downregulation in MCF10A cells leads to enhanced invasion into a 3D matrix. Consistent with these data, ectopic expression of endoglin in MDA-MB-231 cells blocks TGF-β-enhanced cell motility and invasion and reduces lung colonization in an in vivo metastasis model. Unlike endothelial cells, endoglin does not modulate Smad-mediated TGF-β signaling in breast cells but attenuates the cytoskeletal remodeling to impair cell migration and invasion. Importantly, in a large cohort of invasive breast cancers, lack of endoglin expression in the tumor cell compartment correlates with ENG gene methylation and poor clinical outcome.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 50 print issues and online access
$259.00 per year
only $5.18 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Aranda V, Haire T, Nolan ME, Calarco JP, Rosenberg AZ, Fawcett JP et al. (2006). Par6-aPKC uncouples ErbB2 induced disruption of polarized epithelial organization from proliferation control. Nat Cell Biol 8: 1235–1245.
Barbara NP, Wrana JL, Letarte M . (1999). Endoglin is an accessory protein that interacts with the signaling receptor complex of multiple members of the transforming growth factor-beta superfamily. J Biol Chem 274: 584–594.
Bernabeu C, Conley BA, Vary CP . (2007). Novel biochemical pathways of endoglin in vascular cell physiology. J Cell Biochem 102: 1375–1388.
Bernabeu C, Lopez-Novoa JM, Quintanilla M . (2009). The emerging role of TGF-beta superfamily coreceptors in cancer. Biochim Biophys Acta 1792: 954–973.
Bhowmick NA, Ghiassi M, Bakin A, Aakre M, Lundquist CA, Engel ME et al. (2001). Transforming growth factor-beta1 mediates epithelial to mesenchymal transdifferentiation through a RhoA-dependent mechanism. Mol Biol Cell 12: 27–36.
Cheifetz S, Bellon T, Cales C, Vera S, Bernabeu C, Massague J et al. (1992). Endoglin is a component of the transforming growth factor-beta receptor system in human endothelial cells. J Biol Chem 267: 19027–19030.
Conley BA, Koleva R, Smith JD, Kacer D, Zhang D, Bernabeu C et al. (2004). Endoglin controls cell migration and composition of focal adhesions: function of the cytosolic domain. J Biol Chem 279: 27440–27449.
Craft CS, Romero D, Vary CP, Bergan RC . (2007). Endoglin inhibits prostate cancer motility via activation of the ALK2-Smad1 pathway. Oncogene 26: 7240–7250.
Debnath J, Brugge JS . (2005). Modelling glandular epithelial cancers in three-dimensional cultures. Nat Rev Cancer 5: 675–688.
Feng XH, Derynck R . (2005). Specificity and versatility in tgf-beta signaling through Smads. Annu Rev Cell Dev Biol 21: 659–693.
Guerrero-Esteo M, Sanchez-Elsner T, Letamendia A, Bernabeu C . (2002). Extracellular and cytoplasmic domains of endoglin interact with the transforming growth factor-beta receptors I and II. J Biol Chem 277: 29197–29209.
Ikediobi ON, Davies H, Bignell G, Edkins S, Stevens C, O'Meara S et al. (2006). Mutation analysis of 24 known cancer genes in the NCI-60 cell line set. Mol Cancer Ther 5: 2606–2612.
Illingworth RS, Bird AP . (2009). CpG islands—‘a rough guide’. FEBS Lett 583: 1713–1720.
Ishibashi O, Ikegame M, Takizawa F, Yoshizawa T, Moksed MA, Iizawa F et al. (2010). Endoglin is involved in BMP-2-induced osteogenic differentiation of periodontal ligament cells through a pathway independent of Smad-1/5/8 phosphorylation. J Cell Physiol 222: 465–473.
Johnson DW, Berg JN, Baldwin MA, Gallione CJ, Marondel I, Yoon SJ et al. (1996). Mutations in the activin receptor-like kinase 1 gene in hereditary haemorrhagic telangiectasia type 2. Nat Genet 13: 189–195.
Klingbeil P, Natrajan R, Everitt G, Vatcheva R, Marchio C, Palacios J et al. (2010). CD44 is overexpressed in basal-like breast cancers but is not a driver of 11p13 amplification. Breast Cancer Res Treat 120: 95–109.
Lebrin F, Goumans MJ, Jonker L, Carvalho RL, Valdimarsdottir G, Thorikay M et al. (2004). Endoglin promotes endothelial cell proliferation and TGF-beta/ALK1 signal transduction. Embo J 23: 4018–4028.
Lee NY, Blobe GC . (2007). The interaction of endoglin with beta-arrestin2 regulates transforming growth factor-beta-mediated ERK activation and migration in endothelial cells. J Biol Chem 282: 21507–21517.
Massague J . (2008). TGFbeta in Cancer. Cell 134: 215–230.
McAllister KA, Grogg KM, Johnson DW, Gallione CJ, Baldwin MA, Jackson CE et al. (1994). Endoglin, a TGF-beta binding protein of endothelial cells, is the gene for hereditary haemorrhagic telangiectasia type 1. Nat Genet 8: 345–351.
Moustakas A, Heldin CH . (2005). Non-Smad TGF-beta signals. J Cell Sci 118: 3573–3584.
Muthuswamy SK, Li D, Lelievre S, Bissell MJ, Brugge JS . (2001). ErbB2, but not ErbB1, reinitiates proliferation and induces luminal repopulation in epithelial acini. Nat Cell Biol 3: 785–792.
Mythreye K, Blobe GC . (2009). The type III TGF-beta receptor regulates epithelial and cancer cell migration through beta-arrestin2-mediated activation of Cdc42. Proc Natl Acad Sci USA 106: 8221–8226.
Oxmann D, Held-Feindt J, Stark AM, Hattermann K, Yoneda T, Mentlein R . (2008). Endoglin expression in metastatic breast cancer cells enhances their invasive phenotype. Oncogene 27: 3567–3575.
Ozdamar B, Bose R, Barrios-Rodiles M, Wang HR, Zhang Y, Wrana JL . (2005). Regulation of the polarity protein Par6 by TGFbeta receptors controls epithelial cell plasticity. Science 307: 1603–1609.
Padua D, Zhang XH, Wang Q, Nadal C, Gerald WL, Gomis RR et al. (2008). TGFbeta primes breast tumors for lung metastasis seeding through angiopoietin-like 4. Cell 133: 66–77.
Perez-Gomez E, Villa-Morales M, Santos J, Fernandez-Piqueras J, Gamallo C, Dotor J et al. (2007). A role for endoglin as a suppressor of malignancy during mouse skin carcinogenesis. Cancer Res 67: 10268–10277.
Quintanilla M, Ramirez JR, Perez-Gomez E, Romero D, Velasco B, Letarte M et al. (2003). Expression of the TGF-beta coreceptor endoglin in epidermal keratinocytes and its dual role in multistage mouse skin carcinogenesis. Oncogene 22: 5976–5985.
Roberts AB, Wakefield LM . (2003). The two faces of transforming growth factor beta in carcinogenesis. Proc Natl Acad Sci USA 100: 8621–8623.
Rodriguez-Barbero A, Obreo J, Alvarez-Munoz P, Pandiella A, Bernabeu C, Lopez-Novoa JM . (2006). Endoglin modulation of TGF-beta1-induced collagen synthesis is dependent on ERK1/2 MAPK activation. Cell Physiol Biochem 18: 135–142.
Romero D, Terzic A, Conley BA, Craft CS, Jovanovic B, Bergan RC et al. (2010). Endoglin phosphorylation by ALK2 contributes to the regulation of prostate cancer cell migration. Carcinogenesis 31: 359–366.
Sanz-Rodriguez F, Guerrero-Esteo M, Botella LM, Banville D, Vary CP, Bernabeu C . (2004). Endoglin regulates cytoskeletal organization through binding to ZRP-1, a member of the Lim family of proteins. J Biol Chem 279: 32858–32868.
Scharpfenecker M, van Dinther M, Liu Z, van Bezooijen RL, Zhao Q, Pukac L et al. (2007). BMP-9 signals via ALK1 and inhibits bFGF-induced endothelial cell proliferation and VEGF-stimulated angiogenesis. J Cell Sci 120: 964–972.
Scherner O, Meurer SK, Tihaa L, Gressner AM, Weiskirchen R . (2007). Endoglin differentially modulates antagonistic transforming growth factor-beta1 and BMP-7 signaling. J Biol Chem 282: 13934–13943.
Seton-Rogers SE, Lu Y, Hines LM, Koundinya M, LaBaer J, Muthuswamy SK et al. (2004). Cooperation of the ErbB2 receptor and transforming growth factor beta in induction of migration and invasion in mammary epithelial cells. Proc Natl Acad Sci USA 101: 1257–1262.
Shi Y, Massague J . (2003). Mechanisms of TGF-beta signaling from cell membrane to the nucleus. Cell 113: 685–700.
ten Dijke P, Goumans MJ, Pardali E . (2008). Endoglin in angiogenesis and vascular diseases. Angiogenesis 11: 79–89.
Webb DJ, Parsons JT, Horwitz AF . (2002). Adhesion assembly, disassembly and turnover in migrating cells — over and over and over again. Nat Cell Biol 4: E97–100.
Wong VC, Chan PL, Bernabeu C, Law S, Wang LD, Li JL et al. (2008). Identification of an invasion and tumor-suppressing gene, Endoglin (ENG), silenced by both epigenetic inactivation and allelic loss in esophageal squamous cell carcinoma. Int J Cancer 123: 2816–2823.
Zhang YE . (2009). Non-Smad pathways in TGF-beta signaling. Cell Res 19: 128–139.
Acknowledgements
We thank Kay Savage and Suzanne Parry for immunohistochemical staining, Reshma Shah for help with the methylation analysis, Caterina Marchio for microdissection, Senthil Muthuswamy for the MCF10A.ErbB2 cells and Ariad Pharmaceuticals (www.ariad.com/regulationkits) for the AP1510 reagent. This project was funded by Breakthrough Breast Cancer and from NHS funding to the NIHR Biomedical Research Centre, a Marie Curie Intra-European Fellowship (PIEF-GA-2008-221083) and Breast Cancer Research Scotland with support from the Tayside Tissue Bank.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no conflict of interest.
Additional information
Supplementary Information accompanies the paper on the Oncogene website
Supplementary information
Rights and permissions
About this article
Cite this article
Henry, L., Johnson, D., Sarrió, D. et al. Endoglin expression in breast tumor cells suppresses invasion and metastasis and correlates with improved clinical outcome. Oncogene 30, 1046–1058 (2011). https://doi.org/10.1038/onc.2010.488
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/onc.2010.488
Keywords
This article is cited by
-
Identification of novel alternative splicing associated with mastitis disease in Holstein dairy cows using large gap read mapping
BMC Genomics (2022)
-
Endoglin targeting inhibits tumor angiogenesis and metastatic spread in breast cancer
Oncogene (2016)
-
Identification of Endoglin as an epigenetically regulated tumour-suppressor gene in lung cancer
British Journal of Cancer (2015)
-
Activin A Is Anti-Lymphangiogenic in a Melanoma Mouse Model
Journal of Investigative Dermatology (2015)
-
Immunohistochemical expression of endoglin offers a reliable estimation of bone marrow neoangiogenesis in multiple myeloma
Journal of Cancer Research and Clinical Oncology (2015)