Angiogenesis, Metastasis, and the Cellular Microenvironment Hypoxia Stabilizes GAS6/Axl Signaling inMetastatic Prostate Cancer

The receptor tyrosine kinase Axl is overexpressed in a variety of cancers and is known to play a role in proliferation and invasion. Previous data fromour laboratory indicate that Axl and its ligand growth arrest–specific 6 (GAS6)may play a role in establishing metastatic dormancy in the bone marrow microenvironment. In the current study, we found that Axl is highly expressed inmetastatic prostate cancer cell lines PC3 andDU145 and has negligible levels of expression in a nonmetastatic cancer cell line LNCaP. Knockdown of Axl in PC3 and DU145 cells resulted in decreased expression of several mesenchymal markers including Snail, Slug, and N-cadherin, and enhanced expression of the epithelial marker E-cadherin, suggesting that Axl is involved in the epithelial–mesenchymal transition in prostate cancer cells. The Axl-knockdown PC3 and DU145 cells also displayed decreased in vitro migration and invasion. Interestingly, when PC3 andDU145 cells were treated with GAS6, Axl protein levels were downregulated.Moreover,CoCl2, a hypoxiamimicking agent, preventedGAS6-mediated downregulation of Axl in these cell lines. Immunochemical staining of human prostate cancer tissuemicroarrays showed that Axl, GAS6, and hypoxia-inducible factor-1a (Hif-1a; indicator of hypoxia) were all coexpressed in prostate cancer and in bone metastases compared with normal tissues. Together, our studies indicate that Axl plays a crucial role in prostate cancer metastasis and that GAS6 regulates the expression of Axl. Importantly, in a hypoxic tumor microenvironment Axl expression is maintained leading to enhanced signaling.Mol Cancer Res; 10(6); 703–12. 2012 AACR.