Original Article

Laminin-332 cleavage by matriptase alters motility parameters of prostate cancer cells

  1. Manisha Tripathi1,
  2. Alka A. Potdar2,
  3. Hironobu Yamashita1,
  4. Brandy Weidow1,
  5. Peter T. Cummings2,3,
  6. Daniel Kirchhofer4,
  7. Vito Quaranta1,*

Article first published online: 29 JUL 2010

DOI: 10.1002/pros.21233

Issue

The Prostate

The Prostate

Volume 71, Issue 2, pages 184–196, 1 February 2011

Additional Information

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

Author Information

  1. 1

    Department of Cancer Biology, Vanderbilt University Medical Center, Nashville, Tennessee

  2. 2

    Department of Chemical and Biomolecular Engineering, Vanderbilt University, Nashville, Tennessee

  3. 3

    Center for Nanophase Materials Sciences, Oak Ridge National Laboratory, Oak Ridge, Tennessee

  4. 4

    Department of Protein Engineering, Genentech, Inc., South San Francisco, California

*Department of Cancer Biology, Vanderbilt University School of Medicine, 771 Preston Research Building, 2220 Pierce Avenue, Nashville, TN 37232-6840.

Publication History

  1. Issue published online: 22 DEC 2010
  2. Article first published online: 29 JUL 2010
  3. Manuscript Accepted: 18 JUN 2010
  4. Manuscript Received: 15 APR 2010

Funded by

  • National Institutes of Health. Grant Numbers: CA47858-17A2, GM067221-03

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Keywords:

  • laminin-332;
  • matriptase;
  • type II transmembrane serine protease;
  • proteolysis;
  • prostate cancer;
  • cell migration

Abstract

BACKGROUND

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.

METHODS

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.

RESULTS

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.

CONCLUSIONS

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.

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