Research Article| Volume 49, ISSUE 11, P2608-2618, July 2013

Hypoxia inducible factor-1 is activated by transcriptional co-activator with PDZ-binding motif (TAZ) versus WWdomain-containing oxidoreductase (WWOX) in hypoxic microenvironment of bone metastasis from breast cancer

Published:April 08, 2013DOI:


      The hypoxic microenvironment of bone marrow favours the bone metastasis process. Hypoxia inducible factor (HIF)-1α is hallmark for hypoxia, correlating with poor prognosis and radio/chemotherapy resistance of primary-breast carcinoma. For bone metastasis, the molecular mechanisms involved in HIF-1α expression and HIF-1 (α/β heterodimer)-transcription factor activity are scarcely known. We studied the role played by HIF-1 in the cross-talk between neoplastic and supportive-microenvironmental cells. Also, WWdomain-containing oxidoreductase (Wwox) and transcriptional co-activator with PDZ-binding motif (TAZ) were taken into consideration evaluating whether these Hippo-pathway effectors affect bone-metastatic phenotype through HIF-1 activity. Considering bone-metastasis specimens, nuclear HIF-1α–TAZ co-localisation occurred in neoplastic and supportive cells, such as fibroblasts and endotheliocytes. Based on these data, the functional importance was verified using 1833-bone metastatic clone under hypoxia: nuclear HIF-1α and TAZ expression increased and co-immunoprecipitated, activating HIF-1-DNA binding and transactivation. In contrast, Wwox localised at perinuclear level in neoplastic cells of bone metastasis, being almost absent in supportive cells, and Wwox-protein expression diminished in hypoxic-1833 cells. Thus, TAZ regulation of HIF-1 activity might be important for bone-secondary growth, participating in metastasis-stroma cross-talk. Further, TAZ and HIF-1α-protein levels seemed correlated. In fact, blocking cyclooxygenase-2 with NS398 in hypoxic-1833 cells, not only HIF-1α decreased but also molecular-mechanism(s) upstream of the Hippo pathway were triggered: LATS-dependent TAZ phosphorylation seemed responsible for TAZ nucleus/cytoplasm translocation and degradation. In the 1833-xenograft model, NS398 largely prevented the outgrowth of bone-metastatic cells, probably related to remarkable-extracellular matrix assembly. We gained clinical insight into HIF-1α and TAZ as candidate biomarkers for bone avidity, relevant for early-therapeutic intervention against bone metastasis.


      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'


      Subscribe to European Journal of Cancer
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect


        • Lu X.
        • Yan C.H.
        • Yuan M.
        • Wei Y.
        • Hu G.
        • Kang Y.
        In vivo dynamics and distinct function of hypoxia in primary tumor growth and organotropic metastasis of breast cancer.
        Cancer Res. 2010; 70: 3905-3914
        • Semenza G.L.
        Oxygen sensing, homeostasis, and disease.
        N Engl J Med. 2011; 365: 537-547
        • Wong C.C.
        • Gilkes D.M.
        • Zhang H.
        • et al.
        Hypoxia-inducible factor 1 is a master regulator of breast cancer metastatic niche formation.
        Proc Natl Acad Sci USA. 2011; 108: 16369-16374
        • Tian Y.-M.
        • Yeoh K.K.
        • Lee M.K.
        • et al.
        Differential sensitivity of hypoxia inducible factor hydroxylation sites to hypoxia and hydroxylase inhibitors.
        J Biol Chem. 2011; 286: 13041-13051
        • Maroni P.
        • Matteucci E.
        • Luzzati A.
        • Perrucchini G.
        • Bendinelli P.
        • Desiderio M.A.
        Nuclear co-localization and functional interaction of COX-2 and HIF-1α characterize bone metastasis of human breast carcinoma.
        Breast Cancer Res Treat. 2011; 129: 433-450
        • Wilson C.L.
        • Thomsen J.
        • Hoivik D.J.
        • et al.
        Aryl hydrocarbon (Ah) nonresponsiveness in estrogen receptor-negative MDA-MB231 cells is associated with the expression of variant Arnt protein.
        Arch Biochem Biophys. 1997; 346: 65-73
        • Zhong H.
        • De Marzo A.M.
        • Laughner E.
        • et al.
        Overexpression of hypoxia-inducible factor 1 alpha in common human cancers and their metastasis.
        Cancer Res. 1999; 15: 5830-5835
        • Montagner M.
        • Enzo E.
        • Forcato M.
        • et al.
        SHARP1 suppresses breast cancer metastasis by promoting degradation of hypoxia-inducible factors.
        Nature. 2012; 487: 380-384
        • Chen H.H.W.
        • Su W.-C.
        • Lin P.-W.
        • Guo H.-R.
        • Lee W.-Y.
        Hypoxia-inducible factor-1α correlates with Met and metastasis in node-negative breast cancer.
        Breast Cancer Res Treat. 2007; 103: 167-175
        • Gort E.H.
        • Groot A.J.
        • van der Wall E.
        • van Diest P.J.
        • Vooijs M.A.
        Hypoxic regulation of metastasis via hypoxia-inducible factors.
        Curr Mol Med. 2008; 8: 60-67
        • Erler J.T.
        • Bennewith K.L.
        • Cox T.R.
        • et al.
        Hypoxia-induced lysyloxidase is a critical mediator of bone marrow cell recruitment to form the premetastatic niche.
        Cancer Cell. 2009; 15: 35-44
        • Weilbaecher K.N.
        • Guise T.A.
        • McCauley L.K.
        Cancer to bone: a fatal attraction.
        Nat Rev Cancer. 2011; 11: 411-425
        • Hiraga T.
        • Kizaka-Kondoh S.
        • Hirota K.
        • Hiraoka M.
        • Yoneda T.
        Hypoxia and hypoxia-inducible factor-1 expression enhance osteolytic bone metastases of breast cancer.
        Cancer Res. 2007; 67: 4154-4163
        • Matteucci E.
        • Maroni P.
        • Luzzati A.
        • Perrucchini G.
        • Bendinelli P.
        • Desiderio M.A.
        Bone metastatic process of breast cancer involves methylation state affecting E-cadherin expression through TAZ and Wwox nuclear effectors.
        Eur J Cancer. 2013; 49: 231-244
        • Tiwari N.
        • Gheldof A.
        • Tatari M.
        • Christofori G.
        EMT as the ultimate survival mechanism of cancer cells.
        Semin Cancer Biol. 2012; 22: 194-207
        • Cordenonsi M.
        • Zanconato F.
        • Azzolin L.
        • et al.
        The Hippo transducer TAZ confers cancer stem cell-related traits on breast cancer cells.
        Cell. 2011; 147: 759-772
        • Sudol M.
        • Harvey K.F.
        Modularity in the Hippo signaling pathway.
        Trends Biochem Sci. 2010; 35: 627-633
        • Liu C.
        • Huang W.
        • Lei Q.
        Regulation and function of the TAZ transcription co-activator.
        Int J Biochem Mol Biol. 2011; 2: 247-256
        • Aqeilan R.I.
        • Donati V.
        • Gaudino E.
        • et al.
        Association of Wwox with ErbB4 in breast cancer.
        Cancer Res. 2007; 67: 9330-9336
        • Matteucci E.
        • Bendinelli P.
        • Desiderio M.A.
        Nuclear localization of active HGF receptor Met in aggressive MDA-MB231 breast carcinoma cells.
        Carcinogenesis. 2009; 30: 937-945
        • Del Mare S.
        • Salah Z.
        • Aqeilan R.I.
        WWOX: its genomics, partners, and functions.
        J Cell Biochem. 2009; 108: 737-745
        • Muraki C.
        • Ohga N.
        • Hida Y.
        • et al.
        Cyclooxygenase-2 inhibition causes antiangiogenic effects on tumor endothelial and vascular progenitor cells.
        Int J Cancer. 2012; 130: 59-70
        • Hiraga T.
        • Myoui A.
        • Choi M.E.
        • Yoshikawa H.
        • Yoneda T.
        Stimulation of cyclooxygenase-2 expression by bone-derived transforming growth factor-beta enhances bone metastases in breast cancer.
        Cancer Res. 2006; 66: 2067-2073
        • Vrekoussis T.
        • Chaniotis V.
        • Navrozoglou I.
        • et al.
        Image analysis of breast cancer immunohistochemistry-stained sections using ImageJ: an RGB-based model.
        Anticancer Res. 2009; 29: 4995-4998
        • Tacchini L.
        • Dansi P.
        • Matteucci E.
        • Desiderio M.A.
        Hepatocyte growth factor signalling stimulates hypoxia inducible factor-1 (HIF-1) activity in HepG2 hepatoma cells.
        Carcinogenesis. 2001; 9: 1363-1371
        • Bendinelli P.
        • Matteucci E.
        • Maroni P.
        • Desiderio M.A.
        NF-kB activation, dependent on acetylation/deacetylation, contributes to HIF-1 activity and migration of bone metastatic breast cancer cells.
        Mol Cancer Res. 2009; 7: 1328-1341
        • Varelas X.
        • Sakuma R.
        • Samavarchi-Tehrani P.
        • et al.
        TAZ controls Smad nucleocytoplasmic shuttling and regulates human embryonic stem-cell self-renewal.
        Nat Cell Biol. 2008; 10: 837-848
        • Liao J.
        • McCauley L.K.
        Skeletal metastasis: established and emerging roles of parathyroid hormone related protein (PTHrP).
        Cancer Metastasis Rev. 2006; 25: 559-571
        • Li H.
        • Wolfe A.
        • Septer S.
        • et al.
        Deregulation of Hippo kinase signaling in human hepatic malignancies.
        Liver Int. 2012; 32: 38-47
        • Yoneda T.
        • Hiraga T.
        Crosstalk between cancer cells and bone microenvironment in bone metastasis.
        Biochem Biophys Res Commun. 2005; 328: 679-687
        • Kaplan R.M.
        • Riba R.D.
        • Zazharoulis S.
        • et al.
        VEGFR1-positive haemotopoietic bone marrow progenitors initiate the pre-metastatic niche.
        Nature. 2005; 438: 820-827
        • Desiderio M.A.
        Hepatocyte growth factor in invasive growth of carcinomas.
        Cell Mol Life Sci. 2007; 64: 1341-1354
        • Previdi S.
        • Maroni P.
        • Matteucci E.
        • Broggini M.
        • Bendinelli P.
        • Desiderio M.A.
        Interaction between human-breast cancer metastasis and bone microenvironment through activated hepatocyte growth factor/Met and β-catenin/Wnt pathways.
        Eur J Cancer. 2010; 46: 1679-1691
        • Luo D.
        • Wang J.
        • Li J.
        • Post M.
        Mouse snail is a target gene for HIF-1.
        Mol Cancer Res. 2011; 9: 234-245
        • Proulx-Bonneau S.
        • Guezguez A.
        • Annabi B.
        A concerted HIF-1α/MT1-MMP signaling axis regulates the expression of the 3BP2 adaptor protein in hypoxic mesenchymal stromal cells.
        PLoS One. 2011; 6: e21511
        • Liu C.-Y.
        • Zha Z.-Y.
        • Zhou X.
        • et al.
        The Hippo tumor pathway promotes TAZ degradation by phosphorylating a phosphodegron and recruiting the SCFβ-TrCP E3 ligase.
        J Biol Chem. 2010; 285: 37159-37169
        • Kurban G.
        • Duplan E.
        • Ramlal N.
        • et al.
        Collagen matrix assembly is driven by the interaction of von Hippel–Lindau tumor suppressor protein with hydroxylated collagen IV alpha 2.
        Oncogene. 2008; 27: 1004-1012
        • Semenza G.L.
        Targeting HIF-1 for cancer therapy.
        Nat Rev Cancer. 2003; 3: 721-732
        • Melillo G.
        Targeting hypoxia cell signaling foe cancer therapy.
        Cancer Metastasis Rev. 2007; 26: 341-352
        • Joyce J.A.
        • Pollard J.W.
        Microenvironmental regulation of metastasis.
        Nat Rev Cancer. 2009; 9: 239-252
        • Psaila B.
        • Lyden D.
        The metastatic niche: adapting the foreign soil.
        Nat Rev Cancer. 2009; 9: 285-293