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:

Beta4 integrin promotes osteosarcoma metastasis and interacts with ezrin

Oncogene volume 28, pages 3401–3411 (2009)Cite this article

Abstract

The development of pulmonary metastasis is the major cause of death in osteosarcoma, and its molecular basis is poorly understood. In this study, we show that β4 integrin is highly expressed in human osteosarcoma cell lines and tumor samples. Furthermore, highly metastatic MNNG-HOS cells have increased levels of β4 integrin. Suppression of β4 integrin expression by shRNA and disruption of β4 integrin function by transfection of dominant-negative β4 integrin was sufficient to revert this highly metastatic phenotype in the MNNG-HOS model without significantly affecting primary tumor growth. These findings suggest a role for β4 integrin expression in the metastatic phenotype in human osteosarcoma cells. In addition, we identified a previously uncharacterized interaction between β4 integrin and ezrin, a membrane-cytoskeletal linker protein that is implicated in the metastatic behavior of osteosarcoma. The β4 integrin–ezrin interaction appears to be critical for maintenance of β4 integrin expression. These data begin to integrate ezrin and β4 integrin expression into a model of action for the mechanism of osteosarcoma metastases.

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
Figure 4
Figure 5
Figure 6

Similar content being viewed by others

References

  • Andreoli C, Martin M, Le Borgne R, Reggio H, Mangeat P . (1994). Ezrin has properties to self-associate at the plasma membrane. J Cell Sci 107: 2509–2521.

    CAS  PubMed  Google Scholar 

  • Aplin AE, Howe AK, Juliano RL . (1999). Cell adhesion molecules, signal transduction and cell growth. Curr Opin Cell Biol 11: 737–744.

    Article  CAS  Google Scholar 

  • Bertotti A, Comoglio PM, Trusolino L . (2005). Beta4 integrin is a transforming molecule that unleashes Met tyrosine kinase tumorigenesis. Cancer Res 65: 10674–10679.

    Article  CAS  Google Scholar 

  • Bertotti A, Comoglio PM, Trusolino L . (2006). Beta4 integrin activates a Shp2-Src signaling pathway that sustains HGF-induced anchorage-independent growth. J Cell Biol 175: 993–1003.

    Article  CAS  PubMed Central  Google Scholar 

  • Calderwood DA, Fujioka Y, de Pereda JM, Garcia-lvarez B, Nakamoto T, Margolis B et al. (2003). Integrin beta cytoplasmic domain interactions with phosphotyrosine-binding domains: a structural prototype for diversity in integrin signaling. Proc Natl Acad Sci USA 100: 2272–2277.

    Article  CAS  Google Scholar 

  • Calderwood DA, Huttenlocher A, Kiosses WB, Rose DM, Woodside DG, Schwartz MA et al. (2001). Increased filamin binding to beta-integrin cytoplasmic domains inhibits cell migration. Nat Cell Biol 3: 1060–1068.

    Article  CAS  Google Scholar 

  • Calderwood DA, Zent R, Grant R, Rees DJ, Hynes RO, Ginsberg MH . (1999). The Talin head domain binds to integrin beta subunit cytoplasmic tails and regulates integrin activation. J Biol Chem 274: 28071–28074.

    Article  CAS  Google Scholar 

  • Chung J, Bachelder RE, Lipscomb EA, Shaw LM, Mercurio AM . (2002). Integrin (alpha 6 beta 4) regulation of eIF-4E activity and VEGF translation: a survival mechanism for carcinoma cells. J Cell Biol 158: 165–174.

    Article  CAS  PubMed Central  Google Scholar 

  • Gaffney R, Unni KK, Sim FH, Slezak JM, Esther RJ, Bolander ME . (2006). Follow-up study of long-term survivors of osteosarcoma in the prechemotherapy era. Hum Pathol 244: 799–804.

    Google Scholar 

  • Geiger B, Bershadsky A, Pankov R, Yamada KM . (2001). Transmembrane crosstalk between the extracellular matrix--cytoskeleton crosstalk. Nat Rev Mol Cell Biol 2: 793–805.

    Article  CAS  Google Scholar 

  • Giancotti FG, Ruoslahti E . (1999). Integrin signaling. Science 285: 1028–1032.

    Article  CAS  PubMed Central  Google Scholar 

  • Giancotti FG, Tarone G . (2003). Positional control of cell fate through joint integrin/receptor protein kinase signaling. Annu Rev Cell Dev Biol 19: 173–206.

    Article  CAS  PubMed Central  Google Scholar 

  • Grossman HB, Lee C, Bromberg J, Liebert M . (2000). Expression of the alpha6beta4 integrin provides prognostic information in bladder cancer. Oncol Rep 7: 13–16.

    CAS  PubMed  Google Scholar 

  • Guo W, Giancotti FG . (2004). Integrin signalling during tumour progression. Nat Rev Mol Cell Biol 5: 816–826.

    Article  CAS  PubMed Central  Google Scholar 

  • Guo W, Pylayeva Y, Pepe A, Yoshioka T, Muller WJ, Inghirami G et al. (2006). Beta 4 integrin amplifies ErbB2 signaling to promote mammary tumorigenesis. Cell 126: 489–502.

    Article  CAS  Google Scholar 

  • Harris MB, Gieser P, Goorin AM, Ayala A, Shochat SJ, Ferguson WS et al. (1998). Treatment of metastatic osteosarcoma at diagnosis: a Pediatric Oncology Group Study. J Clin Oncol 16: 3641–3648.

    Article  CAS  Google Scholar 

  • Hood JD, Cheresh DA . (2002). Role of integrins in cell invasion and migration. Nat Rev Cancer 2: 91–100.

    Article  Google Scholar 

  • Howe A, Aplin AE, Alahari SK, Juliano RL . (1998). Integrin signaling and cell growth control. Curr Opin Cell Biol 10: 220–231.

    Article  CAS  Google Scholar 

  • Hynes RO . (2002). Integrins: bidirectional, allosteric signaling machines. Cell 110: 673–687.

    Article  CAS  Google Scholar 

  • Kashiwagi H, Schwartz MA, Eigenthaler M, Davis KA, Ginsberg MH, Shattil SJ . (1997). Affinity modulation of platelet integrin alphaIIbbeta3 by beta3-endonexin, a selective binding partner of the beta3 integrin cytoplasmic tail. J Cell Biol 137: 1433–1443.

    Article  CAS  PubMed Central  Google Scholar 

  • Khanna C, Wan X, Bose S, Cassaday R, Olomu O, Mendoza A et al. (2004). The membrane-cytoskeleton linker ezrin is necessary for osteosarcoma metastasis. Nat Med 10: 182–186.

    Article  CAS  PubMed Central  Google Scholar 

  • Kim SY, Lee CH, Midura BV, Yeung C, Mendoza A, Hong SH et al. (2008). Inhibition of the CXCR4/CXCL12 chemokine pathway reduces the development of murine pulmonary metastases. Clin Exp Metastasis 25: 201–211.

    Article  CAS  Google Scholar 

  • Kolanus W, Nagel W, Schiller B, Zeitlmann L, Godar S, Stockinger H et al. (1996). Alpha L beta 2 integrin/LFA-1 binding to ICAM-1 induced by cytohesin-1, a cytoplasmic regulatory molecule. Cell 86: 233–242.

    Article  CAS  Google Scholar 

  • Liu S, Calderwood DA, Ginsberg MH . (2002). Integrin cytoplasmic domain-binding proteins. J Cell Sci 113: 3563–3571.

    Google Scholar 

  • Liu XF, Helman LJ, Yeung C, Bera TK, Lee B, Pastan I . (2000). XAGE-1, a new gene that is frequently expressed in Ewing's sarcoma. Cancer Res 60: 4752–4755.

    CAS  PubMed  Google Scholar 

  • Livak KG, Schmittgen TD . (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods 4: 402–408.

    Article  Google Scholar 

  • Lu S, Simin K, Khan A, Mercurio AM . (2008). Analysis of Integrin {beta}4 Expression in Human Breast Cancer: Association with Basal-like Tumors and Prognostic Significance. Clin Cancer Res 14: 1050–1058.

    Article  CAS  Google Scholar 

  • Mariotti A, Kedeshian PA, Dans M, Curatola AM, Gagnoux-Palacios L, Giancotti FG . (2001). EGF-R signaling through Fyn kinase disrupts the function of integrin alpha6beta4 at hemidesmosomes: role in epithelial cell migration and carcinoma invasion. J Cell Biol 155: 447–458.

    Article  CAS  PubMed Central  Google Scholar 

  • Mercurio AM, Rabinovitz I . (2001). Towards a mechanistic understanding of tumor invasion—lessons from the α6β4integrin. Semin Cancer Biol 11: 129–141.

    Article  CAS  Google Scholar 

  • Meyers PS, Heller G, Healey G, Huvos A, Applewhite A, Sun M et al. (1993). Osteogenic sarcoma with clinically detectable metastasis at initial presentation. J Clin Oncol 11: 449–453.

    Article  CAS  Google Scholar 

  • Nguyen R, Reczek D, Bretscher A . (2001). Hierarchy of merlin and ezrin N- and C-terminal domain interactions in homo- and heterotypic associations and their relationship to binding of scaffolding proteins EBP50 and E3KARP. J Biol Chem 276: 7621–7629.

    Article  CAS  Google Scholar 

  • Nikolopoulos SN, Blaikie P, Yoshioka T, Guo W, Giancotti FG . (2004). Integrin beta4 signaling promotes tumor angiogenesis. Cancer Cell 6: 471–483.

    Article  CAS  Google Scholar 

  • Pfaff M, Du X, Ginsberg MH . (1999). Calpain cleavage of integrin beta cytoplasmic domains. FEBS Lett 460: 17–22.

    Article  CAS  Google Scholar 

  • Rhim JS, Park DK, Arnstein P, Huebner RJ, Weisburgern EK, Nelson-Rees WA . (1975). Transformation of human cells in culture by N-methyl-N′-nitro-N-nitrosoguanidine. Nature 256: 751–753.

    Article  CAS  Google Scholar 

  • Shaw LM . (2001). Identification of insulin receptor substrate 1 (IRS-1) and IRS-2 as signaling intermediates in the alpha6beta4 integrin-dependent activation of phosphoinositide 3-OH kinase and promotion of invasion. Mol Cell Biol 21: 5082–5093.

    Article  CAS  PubMed Central  Google Scholar 

  • Tagliabue E, Ghirelli C, Squicciarini P, Aiello P, Colnaghi MI, Menard S . (1998). Prognostic value of alpha 6 beta 4 integrin expression in breast carcinomas is affected by laminin production from tumor cells. Clin Cancer Res 4: 407–410.

    CAS  PubMed  Google Scholar 

  • Trusolino L, Bertotti A, Comoglio PM . (2001). A signaling adapter function for alpha6beta4 integrin in the control of HGF-dependent invasive growth. Cell 107: 643–654.

    Article  CAS  PubMed Central  Google Scholar 

  • Wan X, Helman LJ . (2003). Levels of PTEN protein modulate Akt phosphorylation on serine 473, but not on threonine 308, in IGF-II-overexpressing rhabdomyosarcomas cells. Oncogene 22: 8205–8211.

    Article  CAS  Google Scholar 

  • Wan X, Mendoza A, Khanna C, Helman LJ . (2005). Rapamycin inhibits ezrin-mediated metastatic behavior in a murine model of osteosarcoma. Cancer Res 65: 2406–2411.

    Article  CAS  PubMed Central  Google Scholar 

Download references

Acknowledgements

We thank Dr AM Mercurio for providing empty vector and β4 mutant (Y1494F), Dr L Trusolino for providing control-shRNA and β4-shRNA, Dr M Arpin for providing GST and GST-ezrin-CT, Dr R Nguyen for providing BSA and ezrin-NT. This research was supported by the Intramural Research Program of the NIH, National Cancer Institute, Center for Cancer Research.

Author information

Authors and Affiliations

  1. Pediatric Oncology Branch, Center for Cancer Research, National Cancer Institute, Bethesda, MD, USA

    X Wan, S Y Kim, L M Guenther, A Mendoza, J Briggs, C Yeung, D Currier, H Zhang, C Mackall, C Khanna & L Helman

  2. Cartilage Biology and Orthopaedics Branch, National Institute of Arthritis and Musculoskeletal and Skin Diseases, NIH, Bethesda, MD, USA

    W-J Li & R S Tuan

  3. Department of Pathology, Emory University, Altanta, GA, USA

    A T Deyrup

Authors
  1. X Wan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. S Y Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. L M Guenther
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. A Mendoza
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. J Briggs
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. C Yeung
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. D Currier
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. H Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. C Mackall
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. W-J Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. R S Tuan
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. A T Deyrup
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. C Khanna
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. L Helman
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to X Wan.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wan, X., Kim, S., Guenther, L. et al. Beta4 integrin promotes osteosarcoma metastasis and interacts with ezrin. Oncogene 28, 3401–3411 (2009). https://doi.org/10.1038/onc.2009.206

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2009.206

Keywords

This article is cited by

Search

Advanced search

Quick links