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:

Axl promotes cell invasion by inducing MMP-9 activity through activation of NF-κB and Brg-1

Oncogene volume 27, pages 4044–4055 (2008)Cite this article

Abstract

Activity of the Axl receptor tyrosine kinase is positively correlated with tumor metastasis; however, its detailed role in the mechanism of tumor invasion is still not completely understood. Here, we show that Axl enhances the expression of matrix metalloproteinase 9 (MMP-9), required for Axl-mediated invasion both in vitro and in vivo. We found that the highly selective MEK1/2 inhibitors U0126 and PD98059, and the expressed dominant-negative form of extracellular signal-regulated kinase (ERK), completely block Axl-mediated MMP-9 activation. In contrast, the phosphatidylinositol 3-kinase inhibitor LY294002 and wortmannin had little effect on activation. Interestingly, however, the Axl ligand Gas6 is not involved in Axl-mediated MMP-9 activation. Mutation of Glu59Axl and Thr77Axl dramatically reduced Gas6–Axl binding but continued to induce MMP-9 activation. In addition, overexpression of Axl-activated ERK and enhanced nuclear factor-κB (NF-κB) transactivation and brahma-related gene-1 (Brg-1) translocation. Exposure to the NF-κB inhibitor silibinin, which inhibits IκBα kinase activity, or overexpression of the dominant-negative mutant IκB and Brg-1 strikingly inhibited Axl-mediated MMP-9 activation. These data indicate that coordination of ERK signaling and NF-κB and Brg-1 activation are indispensable to regulation of Axl-dependent MMP-9 gene transcription. Together with previous data, our results provide a plausible mechanism for Axl-mediated tumor invasion and establish a functional link between the Axl and MMP-9 signaling pathways.

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

  • Banine F, Bartlett C, Gunawardena R, Muchardt C, Yaniv M, Knudsen ES et al. (2005). SWI/SNF chromatin-remodeling factors induce changes in DNA methylation to promote transcriptional activation. Cancer Res 65: 3542–3547.

    Article  CAS  Google Scholar 

  • Bittner M, Meltzer P, Chen Y, Jiang Y, Seftor E, Hendrix M et al. (2000). Molecular classification of cutaneous malignant melanoma by gene expression profiling. Nature 406: 536–540.

    Article  CAS  Google Scholar 

  • Budagian V, Bulanova E, Orinska Z, Thon L, Mamat U, Bellosta P et al. (2005). A promiscuous liaison between IL-15 receptor and Axl receptor tyrosine kinase in cell death control. EMBO J 24: 4260–4270.

    Article  CAS  Google Scholar 

  • Chung BI, Malkowicz SB, Nguyen TB, Libertino JA, McGarvey TW . (2003). Expression of the proto-oncogene Axl in renal cell carcinoma. DNA Cell Biol 22: 533–540.

    Article  CAS  Google Scholar 

  • Craven RJ, Xu LH, Weiner TM, Fridell YW, Dent GA, Srivastava S et al. (1995). Receptor tyrosine kinases expressed in metastatic colon cancer. Int J Cancer 60: 791–797.

    Article  CAS  Google Scholar 

  • Demarchi F, Verardo R, Varnum B, Brancolini C, Schneider C . (2001). Gas6 anti-apoptotic signaling requires NF-kappa B activation. J Biol Chem 276: 31738–31744.

    Article  CAS  Google Scholar 

  • Dhanalakshmi S, Singh RP, Agarwal C, Agarwal R . (2002). Silibinin inhibits constitutive and TNFalpha-induced activation of NF-kappaB and sensitizes human prostate carcinoma DU145 cells to TNFalpha-induced apoptosis. Oncogene 21: 1759–1767.

    Article  CAS  Google Scholar 

  • DiRenzo J, Shang Y, Phelan M, Sif S, Myers M, Kingston R et al. (2000). BRG-1 is recruited to estrogen-responsive promoters and cooperates with factors involved in histone acetylation. Mol Cell Biol 20: 7541–7549.

    Article  CAS  Google Scholar 

  • Fingleton B . (2006). Matrix metalloproteinases: roles in cancer and metastasis. Front Biosci 11: 479–491.

    Article  CAS  Google Scholar 

  • Fridell YW, Jin Y, Quilliam LA, Burchert A, McCloskey P, Spizz G et al. (1996). Differential activation of the Ras/extracellular-signal-regulated protein kinase pathway is responsible for the biological consequences induced by the Axl receptor tyrosine kinase. Mol Cell Biol 16: 135–145.

    Article  CAS  Google Scholar 

  • Genersch E, Hayess K, Neuenfeld Y, Haller H . (2000). Sustained ERK phosphorylation is necessary but not sufficient for MMP-9 regulation in endothelial cells: involvement of Ras-dependent and -independent pathways. J Cell Sci 113 (Part 23): 4319–4330.

    CAS  PubMed  Google Scholar 

  • Hafizi S, Dahlback B . (2006). Signalling and functional diversity within the Axl subfamily of receptor tyrosine kinases. Cytokine Growth Factor Rev 17: 295–304.

    Article  CAS  Google Scholar 

  • Hasanbasic I, Cuerquis J, Varnum B, Blostein MD . (2004). Intracellular signaling pathways involved in Gas6-Axl-mediated survival of endothelial cells. Am J Physiol Heart Circ Physiol 287: H1207–H1213.

    Article  CAS  Google Scholar 

  • Holland SJ, Powell MJ, Franci C, Chan EW, Friera AM, Atchison RE et al. (2005). Multiple roles for the receptor tyrosine kinase axl in tumor formation. Cancer Res 65: 9294–9303.

    Article  CAS  Google Scholar 

  • Ito T, Ito M, Naito S, Ohtsuru A, Nagayama Y, Kanematsu T et al. (1999). Expression of the Axl receptor tyrosine kinase in human thyroid carcinoma. Thyroid 9: 563–567.

    Article  CAS  Google Scholar 

  • Jacob AN, Kalapurakal J, Davidson WR, Kandpal G, Dunson N, Prashar Y et al. (1999). A receptor tyrosine kinase, UFO/Axl, and other genes isolated by a modified differential display PCR are overexpressed in metastatic prostatic carcinoma cell line DU145. Cancer Detect Prev 23: 325–332.

    Article  CAS  Google Scholar 

  • Jin S, Lu D, Ye S, Ye H, Zhu L, Feng Z et al. (2005). A simplified probe preparation for ELISA-based NF-kappaB activity assay. J Biochem Biophys Methods 65: 20–29.

    Article  CAS  Google Scholar 

  • Lakka SS, Jasti SL, Gondi C, Boyd D, Chandrasekar N, Dinh DH et al. (2002). Downregulation of MMP-9 in ERK-mutated stable transfectants inhibits glioma invasion in vitro. Oncogene 21: 5601–5608.

    Article  CAS  Google Scholar 

  • Lay JD, Hong CC, Huang JS, Yang YY, Pao CY, Liu CH et al. (2007). Sulfasalazine suppresses drug resistance and invasiveness of lung adenocarcinoma cells expressing AXL. Cancer Res 67: 3878–3887.

    Article  CAS  Google Scholar 

  • Liu Y, Denlinger CE, Rundall BK, Smith PW, Jones DR . (2006). Suberoylanilide hydroxamic acid induces Akt-mediated phosphorylation of p300, which promotes acetylation and transcriptional activation of RelA/p65. J Biol Chem 281: 31359–31368.

    Article  CAS  Google Scholar 

  • Ma Z, Shah RC, Chang MJ, Benveniste EN . (2004). Coordination of cell signaling, chromatin remodeling, histone modifications, and regulator recruitment in human matrix metalloproteinase 9 gene transcription. Mol Cell Biol 24: 5496–5509.

    Article  CAS  Google Scholar 

  • Melaragno MG, Fridell YW, Berk BC . (1999). The Gas6/Axl system: a novel regulator of vascular cell function. Trends Cardiovasc Med 9: 250–253.

    Article  CAS  Google Scholar 

  • Meric F, Lee WP, Sahin A, Zhang H, Kung HJ, Hung MC . (2002). Expression profile of tyrosine kinases in breast cancer. Clin Cancer Res 8: 361–367.

    CAS  PubMed  Google Scholar 

  • Nakano T, Tani M, Ishibashi Y, Kimura K, Park YB, Imaizumi N et al. (2003). Biological properties and gene expression associated with metastatic potential of human osteosarcoma. Clin Exp Metastasis 20: 665–674.

    Article  CAS  Google Scholar 

  • Nemoto T, Ohashi K, Akashi T, Johnson JD, Hirokawa K . (1997). Overexpression of protein tyrosine kinases in human esophageal cancer. Pathobiology 65: 195–203.

    Article  CAS  Google Scholar 

  • Neubauer A, O'Bryan JP, Fiebeler A, Schmidt C, Huhn D, Liu ET . (1993). Axl, a novel receptor tyrosine kinase isolated from chronic myelogenous leukemia. Semin Hematol 30: 34.

    CAS  PubMed  Google Scholar 

  • Roberts CW, Orkin SH . (2004). The SWI/SNF complex—chromatin and cancer. Nat Rev Cancer 4: 133–142.

    Article  CAS  Google Scholar 

  • Sasaki T, Knyazev PG, Clout NJ, Cheburkin Y, Gohring W, Ullrich A et al. (2006). Structural basis for Gas6-Axl signalling. EMBO J 25: 80–87.

    Article  CAS  Google Scholar 

  • Shieh YS, Lai CY, Kao YR, Shiah SG, Chu YW, Lee HS et al. (2005). Expression of axl in lung adenocarcinoma and correlation with tumor progression. Neoplasia 7: 1058–1064.

    Article  CAS  Google Scholar 

  • Singh RA, Zhang JZ . (2004). Differential activation of ERK, p38, and JNK required for Th1 and Th2 deviation in myelin-reactive T cells induced by altered peptide ligand. J Immunol 173: 7299–7307.

    Article  CAS  Google Scholar 

  • Sun W, Fujimoto J, Tamaya T . (2004). Coexpression of Gas6/Axl in human ovarian cancers. Oncology 66: 450–457.

    Article  CAS  Google Scholar 

  • Tai KY, Shiah SG, Shieh YS, Kao YR, Chi CY, Huang E et al. (2007). DNA methylation and histone modification regulate silencing of epithelial cell adhesion molecule for tumor invasion and progression. Oncogene 26: 3989–3997.

    Article  CAS  Google Scholar 

  • Vajkoczy P, Knyazev P, Kunkel A, Capelle HH, Behrndt S, von Tengg-Kobligk H et al. (2006). Dominant-negative inhibition of the Axl receptor tyrosine kinase suppresses brain tumor cell growth and invasion and prolongs survival. Proc Natl Acad Sci USA 103: 5799–5804.

    Article  CAS  Google Scholar 

  • van Ginkel PR, Gee RL, Shearer RL, Subramanian L, Walker TM, Albert DM et al. (2004). Expression of the receptor tyrosine kinase Axl promotes ocular melanoma cell survival. Cancer Res 64: 128–134.

    Article  CAS  Google Scholar 

  • Wang W . (2003). The SWI/SNF family of ATP-dependent chromatin remodelers: similar mechanisms for diverse functions. Curr Top Microbiol Immunol 274: 143–169.

    CAS  PubMed  Google Scholar 

  • Wu CW, Li AF, Chi CW, Lai CH, Huang CL, Lo SS et al. (2002). Clinical significance of AXL kinase family in gastric cancer. Anticancer Res 22: 1071–1078.

    CAS  PubMed  Google Scholar 

  • Yao J, Xiong S, Klos K, Nguyen N, Grijalva R, Li P et al. (2001). Multiple signaling pathways involved in activation of matrix metalloproteinase-9 (MMP-9) by heregulin-beta1 in human breast cancer cells. Oncogene 20: 8066–8074.

    Article  CAS  Google Scholar 

  • Zeigler ME, Chi Y, Schmidt T, Varani J . (1999). Role of ERK and JNK pathways in regulating cell motility and matrix metalloproteinase 9 production in growth factor-stimulated human epidermal keratinocytes. J Cell Physiol 180: 271–284.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This study was supported by the National Health Research Institutes (Zhunan, Taiwan) grant no. 96A1-CASP01-014 (to C-W Wu). We thank Dr Christian Muchardt (Institute Pasteur, Paris) and Dr Weidong Wang (National Institute on Aging, Baltimore) for providing the BJ-5-Brg-1 and BJ-5-Brg-1(K798R) plasmids; Dr Anthony N Imbalzano for contacting Dr Muchardt and Dr Wang and Dr Jian Jian Li (Purdue University, Indiana) for providing the pCEP4–ERK1(K71R), pCEP4–ERK2(K52R) and pEGFP–IκBα(S32A, S36A) plasmids.

Author information

Authors and Affiliations

  1. Institute of Cancer Research, National Health Research Institutes, Miaoli County, Taiwan

    K-Y Tai, C-S Lee, S-G Shiah & C-W Wu

  2. School of Dentistry, National Defense Medical Center, Taipei, Taiwan

    Y-S Shieh

Authors
  1. K-Y Tai
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Y-S Shieh
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. C-S Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. S-G Shiah
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. C-W Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to S-G Shiah or C-W Wu.

Additional information

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

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tai, KY., Shieh, YS., Lee, CS. et al. Axl promotes cell invasion by inducing MMP-9 activity through activation of NF-κB and Brg-1. Oncogene 27, 4044–4055 (2008). https://doi.org/10.1038/onc.2008.57

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/onc.2008.57

Keywords

This article is cited by

Search

Advanced search

Quick links