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 (Hsp90α) was identified and shown to activate matrix metalloproteinase 2 (MMP-2). The mechanism of MMP-2 activation by Hsp90α is unknown. Intracellular Hsp90α commonly functions with a complex of co-chaperones, leading to our hypothesis that Hsp90α functions similarly outside of the cell. In this study, we show that a complex of co-chaperones outside of breast Cancer cells assists Hsp90α 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 Hsp90α in vitro and in breast Cancer conditioned media. These co-chaperones also increase the association of Hsp90α and MMP-2 in vitro. This co-chaperone complex enhances Hsp90α-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 Hsp90α 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.