Laminin-332 cleavage by matriptase alters motility parameters of prostate cancer cells
Article first published online: 29 JUL 2010
Copyright © 2010 Wiley-Liss, Inc.
Volume 71, Issue 2, pages 184–196, 1 February 2011
How to Cite
Tripathi, M., Potdar, A. A., Yamashita, H., Weidow, B., Cummings, P. T., Kirchhofer, D. and Quaranta, V. (2011), Laminin-332 cleavage by matriptase alters motility parameters of prostate cancer cells. Prostate, 71: 184–196. doi: 10.1002/pros.21233
- Issue published online: 22 DEC 2010
- Article first published online: 29 JUL 2010
- Manuscript Accepted: 18 JUN 2010
- Manuscript Received: 15 APR 2010
- National Institutes of Health. Grant Numbers: CA47858-17A2, GM067221-03
- type II transmembrane serine protease;
- prostate cancer;
- cell migration
Matriptase, a type II transmembrane serine protease, has been linked to initiation and promotion of epidermal carcinogenesis in a murine model, suggesting that deregulation of its role in epithelia contributes to transformation. In human prostate cancer, matriptase expression correlates with progression. It is therefore of interest to determine how matriptase may contribute to epithelial neoplastic progression. One approach for studying this is to identify potential matriptase substrates involved in epithelial integrity and/or transformation like the extracellular matrix macromolecule, laminin-332 (Ln-332), which is found in the basement membrane of many epithelia, including prostate. Proteolytic processing of Ln-332 regulates cell motility of both normal and transformed cells, which has implications in cancer progression.
In vitro cleavage experiments were performed with purified Ln-332 protein and matriptase. Western blotting, enzyme inhibition assays, and mass spectrometry were used to confirm cleavage events. Matriptase overexpressing LNCaP prostate cancer cells were generated and included in Transwell migration assays and single cell motility assays, along with other prostate cells.
We report that matriptase proteolytically cleaves Ln-332 in the β3 chain. Substrate specificity was confirmed by blocking cleavage with the matriptase inhibitor, Kunitz domain-1. Transwell migration assays showed that DU145 cell motility was significantly enhanced when plated on matriptase-cleaved Ln-332. Similarly, Transwell migration of matriptase-overexpressing LNCaP cells was significantly increased on Ln-332 and, as determined by live single-cell microscopy, two motility parameters of this cell line, speed and directional persistence, were also higher.
Proteolytic processing of Ln-332 by matriptase enhances speed and directional persistence of prostate cancer cells. Prostate 71: 184–196, 2011. © 2010 Wiley-Liss, Inc.