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.
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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.
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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).
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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
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DOI: https://doi.org/10.1038/onc.2008.57
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