1. ¹Institute of Cancer Research, National Health Research Institutes, Miaoli County, Taiwan
2. ²School of Dentistry, National Defense Medical Center, Taipei, Taiwan

Correspondence: Dr C-W Wu and Dr Shine-Gwo Shiah, Institute of Cancer Research, National Health Research Institutes, 35, Keyan Road, Zhunan Town, Miaoli County 350, Taiwan. E-mails: ken@nhri.org.tw and 920114@nhri.org.tw

Received 19 October 2007; Revised 4 January 2008; Accepted 1 February 2008; Published online 17 March 2008.

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 Glu59^Axl and Thr77^Axl dramatically reduced Gas6–Axl binding but continued to induce MMP-9 activation. In addition, overexpression of Axl-activated ERK and enhanced nuclear factor-B (NF-B) transactivation and brahma-related gene-1 (Brg-1) translocation. Exposure to the NF-B inhibitor silibinin, which inhibits IB kinase activity, or overexpression of the dominant-negative mutant IB and Brg-1 strikingly inhibited Axl-mediated MMP-9 activation. These data indicate that coordination of ERK signaling and NF-B 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.