Extracellular Heat Shock Protein (Hsp) 70 and Hsp90α Assist in Matrix Metalloproteinase-2 Activation and Breast Cancer Cell Migration and Invasion

Breast cancer is second only to lung cancer in cancer-related deaths in women, and the majority of these deaths are caused by metastases. Obtaining a better understanding of migration and invasion, two early steps in metastasis, is critical for the development of treatments that inhibit breast cancer metastasis. In a functional proteomic screen for proteins required for invasion, extracellular heat shock protein 90 alpha (Hsp90 alpha) was identified and shown to activate matrix metalloproteinase 2 (MMP-2). The mechanism of MMP-2 activation by Hsp90a is unknown. Intracellular Hsp90a commonly functions with a complex of co- chaperones, leading to our hypothesis that Hsp90a functions similarly outside of the cell. In this study, we show that a complex of co- chaperones outside of breast cancer cells assists Hsp90a mediated activation of MMP-2. We demonstrate that the co- chaperones Hsp70, Hop, Hsp40, and p23 are present outside of breast cancer cells and co-immunoprecipitate with Hsp90 alpha in vitro and in breast cancer conditioned media. These co- chaperones also increase the association of Hsp90 alpha and MMP-2 in vitro. This co-chaperone complex enhances Hsp90 alpha-mediated activation of MMP-2 in vitro, while inhibition of Hsp70 in conditioned media reduces this activation and decreases cancer cell migration and invasion. Together, these findings support a model in which MMP-2 activation by an extracellular co- chaperone complex mediated by Hsp90 alpha increases breast cancer cell migration and invasion. Our studies provide insight into a novel pathway for MMP-2 activation and suggest Hsp70 as an additional extracellular target for anti-metastatic drug development.