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
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
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.
Aplin AE, Howe AK, Juliano RL . (1999). Cell adhesion molecules, signal transduction and cell growth. Curr Opin Cell Biol 11: 737–744.
Bertotti A, Comoglio PM, Trusolino L . (2005). Beta4 integrin is a transforming molecule that unleashes Met tyrosine kinase tumorigenesis. Cancer Res 65: 10674–10679.
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.
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.
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.
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.
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.
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.
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.
Giancotti FG, Ruoslahti E . (1999). Integrin signaling. Science 285: 1028–1032.
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.
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.
Guo W, Giancotti FG . (2004). Integrin signalling during tumour progression. Nat Rev Mol Cell Biol 5: 816–826.
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.
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.
Hood JD, Cheresh DA . (2002). Role of integrins in cell invasion and migration. Nat Rev Cancer 2: 91–100.
Howe A, Aplin AE, Alahari SK, Juliano RL . (1998). Integrin signaling and cell growth control. Curr Opin Cell Biol 10: 220–231.
Hynes RO . (2002). Integrins: bidirectional, allosteric signaling machines. Cell 110: 673–687.
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.
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.
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.
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.
Liu S, Calderwood DA, Ginsberg MH . (2002). Integrin cytoplasmic domain-binding proteins. J Cell Sci 113: 3563–3571.
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.
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.
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.
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.
Mercurio AM, Rabinovitz I . (2001). Towards a mechanistic understanding of tumor invasion—lessons from the α6β4integrin. Semin Cancer Biol 11: 129–141.
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.
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.
Nikolopoulos SN, Blaikie P, Yoshioka T, Guo W, Giancotti FG . (2004). Integrin beta4 signaling promotes tumor angiogenesis. Cancer Cell 6: 471–483.
Pfaff M, Du X, Ginsberg MH . (1999). Calpain cleavage of integrin beta cytoplasmic domains. FEBS Lett 460: 17–22.
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.
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.
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.
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.
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.
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.
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
Corresponding author
Rights 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
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/onc.2009.206
Keywords
This article is cited by
-
Adhesion, metastasis, and inhibition of cancer cells: a comprehensive review
Molecular Biology Reports (2024)
-
Differential recruitment of CD44 isoforms by ErbB ligands reveals an involvement of CD44 in breast cancer
Oncogene (2018)
-
Antitumor activity and expression profiles of genes induced by sulforaphane in human melanoma cells
European Journal of Nutrition (2018)
-
Significance of MTA1 in the molecular characterization of osteosarcoma
Cancer and Metastasis Reviews (2014)
-
ITGA3 and ITGB4 expression biomarkers estimate the risks of locoregional and hematogenous dissemination of oral squamous cell carcinoma
BMC Cancer (2013)