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 Paper
  • Published:

Tyrosine phosphorylation of paxillin affects the metastatic potential of human osteosarcoma

Oncogene volume 24, pages 4754–4764 (2005)Cite this article

Abstract

To acquire information on signal alteration corresponding to the changes in metastatic potential, we analysed protein tyrosine phosphorylation of low- and high-metastatic human osteosarcoma HuO9 sublines, which were recently established as the first metastatic model of human osteosarcoma. Tyrosine phosphorylation of proteins around 60, 70, and 120–130 kDa was enhanced in high-metastatic sublines. Among these proteins, the protein around 70 kDa, which was most remarkably phosphorylated, was identified as paxillin, a scaffold protein in integrin signaling. Activity of Src family kinase correlated well with metastatic potential, and a Src family kinase inhibitor, PP2, not only abolished tyrosine phosphorylation of paxillin but also impaired the motility of high-metastatic sublines. The expression of paxillin was also elevated in high-metastatic sublines, and knocking down of paxillin expression by RNAi method resulted in attenuated motility of high-metastatic cells. We also demonstrated that the phosphorylated form of paxillin is essential for the migration-promoting effect in human osteosarcoma. These findings suggest that enhanced activity of Src family kinases and overexpression of paxillin synergistically contribute to the high metastatic potential of human osteosarcoma through the hyperphosphorylation of paxillin.

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

Abbreviations

BSA:

bovine serum albumin

Cas:

Crk-associated substrate

FAK:

focal adhesion kinase

FBS:

fetal bovine serum

PBS:

phosphate-buffered saline

PP2:

4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine

PP3:

4-amino-7-phenylpyrazol [3,4-d]pyrimidine

RNAi:

RNA interference

SH2:

Src homology 2 domain

siRNA:

short interfering RNA

References

  • Batanian JR, Cavalli LR, Aldosari NM, Ma E, Sotelo-Avila C, Ramos MB, Rone JD, Thorpe CM and Haddad BR . (2002). Mol. Pathol., 55, 389–393.

  • Burnham MR, Bruce-Staskal PJ, Harte MT, Weidow CL, Ma A, Weed SA and Bouton AH . (2000). Mol. Cell. Biol., 20, 5865–5878.

  • Burridge K, Turner CE and Romer LH . (1992). J. Cell. Biol., 119, 893–903.

  • Huang J, Asawa T, Takato T and Sakai R . (2003). J. Biol. Chem., 278, 48367–48376.

  • Huth JF and Eilber FR . (1989). Arch. Surg., 124, 122–126.

  • Iwaya K, Ogawa H, Kuroda M, Izumi M, Ishida T and Mukai K . (2003). Clin. Exp. Metast., 20, 525–529.

  • Khanna C, Khan J, Nguyen P, Prehn J, Caylor J, Yeung C, Trepel J, Meltzer P and Helman L . (2001). Cancer Res., 61, 3750–3759.

  • Khanna C, Wan X, Bose S, Cassaday R, Olomu O, Mendoza A, Yeung C, Gorlick R, Hewitt SM and Helman LJ . (2004). Nat. Med., 10, 182–186.

  • Kimura K, Nakano T, Park YB, Tani M, Tsuda H, Beppu Y, Moriya H and Yokota J . (2002). Clin. Exp. Metast., 19, 477–485.

  • Kiyokawa E, Hashimoto Y, Kobayashi S, Sugimura H, Kurata T and Matsuda M . (1998). Genes Dev., 12, 3331–3336.

  • Klinghoffer RA, Sachsenmaier C, Cooper JA and Soriano P . (1999). EMBO J., 18, 2459–2471.

  • Lau CC, Harris CP, Lu XY, Perlaky L, Gogineni S, Chintagumpala M, Hicks J, Johnson ME, Davino NA, Huvos AG, Meyers PA, Healy JH, Gorlick R and Rao PH . (2004). Genes Chromosomes Cancer, 39, 11–21.

  • Lewis JM and Schwartz MA . (1998). J. Biol. Chem., 273, 14225–14230.

  • Marchetti D, Parikh N, Sudol M and Gallick GE . (1998). Oncogene, 16, 3253–3260.

  • Meyers PA, Heller G, Healey JH, Huvos A, Applewhite A, Sun M and LaQuaglia M . (1993). J. Clin. Oncol., 11, 449–453.

  • Nakamoto T, Sakai R, Ozawa K, Yazaki Y and Hirai H . (1996). J. Biol. Chem., 271, 8959–8965.

  • Nakano T, Tani M, Ishibashi Y, Kimura K, Park YB, Imaizumi N, Tsuda H, Aoyagi K, Sasaki H, Ohwada S and Yokota J . (2003). Clin. Exp. Metast., 20, 665–674.

  • Pawson T . (2004). Cell, 116, 191–203.

  • Petit V, Boyer B, Lentz D, Turner CE, Thiery JP and Valles AM . (2000). J. Cell. Biol., 148, 957–970.

  • Sakai R, Iwamatsu A, Hirano N, Ogawa S, Tanaka T, Mano H, Yazaki Y and Hirai H . (1994). EMBO J., 13, 3748–3756.

  • Schaller MD . (2001). Oncogene, 20, 6459–6472.

  • Schaller MD, Hildebrand JD, Shannon JD, Fox JW, Vines RR and Parsons JT . (1994). Mol. Cell. Biol., 14, 1680–1688.

  • Schaller MD and Parsons JT . (1995). Mol. Cell. Biol., 15, 2635–2645.

  • Schlaepfer DD, Broome MA and Hunter T . (1997). Mol. Cell. Biol., 17, 1702–1713.

  • Squire JA, Pei J, Marrano P, Beheshti B, Bayani J, Lim G, Moldovan L and Zielenska M . (2003). Genes Chromosomes Cancer, 38, 215–225.

  • Talamonti MS, Roh MS, Curley SA and Gallick GE . (1993). J. Clin. Invest., 91, 53–60.

  • Tsubouchi A, Sakakura J, Yagi R, Mazaki Y, Schaefer E, Yano H and Sabe H . (2002). J. Cell. Biol., 159, 673–683.

  • Tsuchiya H, Kanazawa Y, Abdel-Wanis ME, Asada N, Abe S, Isu K, Sugita T and Tomita K . (2002). J. Clin. Oncol., 20, 3470–3477.

  • Turner CE . (2000). Nat. Cell. Biol., 2, E231–E236.

  • Ward WG, Mikaelian K, Dorey F, Mirra JM, Sassoon A, Holmes EC, Eilber FR and Eckardt JJ . (1994). J. Clin. Oncol., 12, 1849–1858.

  • Yano H, Uchida H, Iwasaki T, Mukai M, Akedo H, Nakamura K, Hashimoto S and Sabe H . (2000). Proc. Natl. Acad. Sci. USA, 97, 9076–9081.

  • Yeatman TJ . (2004). Nat. Rev. Cancer, 4, 470–480.

Download references

Acknowledgements

We are grateful to Dr Y Sawada (Department of Biological Sciences, Columbia University) for donating paxillin cDNA. KA is an awardee of the Research Resident Fellowship from the Foundation for Promoting of Cancer Research (Japan) for the 3rd Term Comprehensive 10-Year-Strategy for Cancer Control. This study was supported by the Program for Promotion of Fundamental Studies in Health Science of Pharmaceuticals and Medical Devices Agency (PMDA).

Author information

Authors and Affiliations

  1. Growth Factor Division, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, 104-0045, Tokyo, Japan

    Kotaro Azuma, Masamitsu Tanaka, Takamasa Uekita & Ryuichi Sakai

  2. Department of Geriatric Medicine, Graduate School of Medicine, The University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, 113-8655, Tokyo, Japan

    Kotaro Azuma, Satoshi Inoue & Yasuyoshi Ouchi

  3. Division of Biology, National Cancer Center Research Institute, 5-1-1 Tsukiji, Chuo-ku, 104-0045, Tokyo, Japan

    Jun Yokota

Authors
  1. Kotaro Azuma
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Masamitsu Tanaka
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Takamasa Uekita
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Satoshi Inoue
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jun Yokota
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yasuyoshi Ouchi
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Ryuichi Sakai
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ryuichi Sakai.

Additional information

Supplementary Information accompanies the paper on Oncogene website (http://www.nature.com/onc)

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Azuma, K., Tanaka, M., Uekita, T. et al. Tyrosine phosphorylation of paxillin affects the metastatic potential of human osteosarcoma. Oncogene 24, 4754–4764 (2005). https://doi.org/10.1038/sj.onc.1208654

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.onc.1208654

Keywords

This article is cited by

Search

Advanced search

Quick links