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

Manish Tripathi, Alka A. Potdar, Hironobu Yamashita, Brandy Weidow, Peter T. Cummings, Daniel Kirchhofer, Vito Quaranta

Research output: Contribution to journalArticle

20 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)184-196
Number of pages13
JournalProstate
Volume71
Issue number2
DOIs
StatePublished - Feb 1 2011
Externally publishedYes

Fingerprint

Prostatic Neoplasms
Cell Movement
Prostate
kalinin
matriptase
Epithelium
Cell Migration Assays
Enzyme Assays
Serine Proteases
Substrate Specificity
Basement Membrane
Extracellular Matrix
Microscopy
Mass Spectrometry
Carcinogenesis
Western Blotting
Cell Line

All Science Journal Classification (ASJC) codes

  • Oncology
  • Urology

Cite this

Tripathi, M., Potdar, A. A., Yamashita, H., Weidow, B., Cummings, P. T., Kirchhofer, D., & Quaranta, V. (2011). Laminin-332 cleavage by matriptase alters motility parameters of prostate cancer cells. Prostate, 71(2), 184-196. https://doi.org/10.1002/pros.21233

Laminin-332 cleavage by matriptase alters motility parameters of prostate cancer cells. / Tripathi, Manish; Potdar, Alka A.; Yamashita, Hironobu; Weidow, Brandy; Cummings, Peter T.; Kirchhofer, Daniel; Quaranta, Vito.

In: Prostate, Vol. 71, No. 2, 01.02.2011, p. 184-196.

Research output: Contribution to journalArticle

Tripathi, M, Potdar, AA, Yamashita, H, Weidow, B, Cummings, PT, Kirchhofer, D & Quaranta, V 2011, 'Laminin-332 cleavage by matriptase alters motility parameters of prostate cancer cells', Prostate, vol. 71, no. 2, pp. 184-196. https://doi.org/10.1002/pros.21233
Tripathi M, Potdar AA, Yamashita H, Weidow B, Cummings PT, Kirchhofer D et al. Laminin-332 cleavage by matriptase alters motility parameters of prostate cancer cells. Prostate. 2011 Feb 1;71(2):184-196. https://doi.org/10.1002/pros.21233
Tripathi, Manish ; Potdar, Alka A. ; Yamashita, Hironobu ; Weidow, Brandy ; Cummings, Peter T. ; Kirchhofer, Daniel ; Quaranta, Vito. / Laminin-332 cleavage by matriptase alters motility parameters of prostate cancer cells. In: Prostate. 2011 ; Vol. 71, No. 2. pp. 184-196.
@article{6ea64f0488e547d8a252863c54d02328,
title = "Laminin-332 cleavage by matriptase alters motility parameters of prostate cancer cells",
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.",
author = "Manish Tripathi and Potdar, {Alka A.} and Hironobu Yamashita and Brandy Weidow and Cummings, {Peter T.} and Daniel Kirchhofer and Vito Quaranta",
year = "2011",
month = "2",
day = "1",
doi = "10.1002/pros.21233",
language = "English (US)",
volume = "71",
pages = "184--196",
journal = "Prostate",
issn = "0270-4137",
publisher = "Wiley-Liss Inc.",
number = "2",

}

TY - JOUR

T1 - Laminin-332 cleavage by matriptase alters motility parameters of prostate cancer cells

AU - Tripathi, Manish

AU - Potdar, Alka A.

AU - Yamashita, Hironobu

AU - Weidow, Brandy

AU - Cummings, Peter T.

AU - Kirchhofer, Daniel

AU - Quaranta, Vito

PY - 2011/2/1

Y1 - 2011/2/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=78650977062&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=78650977062&partnerID=8YFLogxK

U2 - 10.1002/pros.21233

DO - 10.1002/pros.21233

M3 - Article

VL - 71

SP - 184

EP - 196

JO - Prostate

JF - Prostate

SN - 0270-4137

IS - 2

ER -