TMPRSS2 is a type II transmembrane-bound serine protease that has gained interest owing to its highly localized expression in the prostate and its overexpression in neoplastic prostate epithelium. Once activated, the serine protease domain of TMPRSS2 is released from the cell surface into the extracellular space. PAR (protease-activated receptor)-2 belongs to a family of G-protein-coupled receptors (PAR-1–4) that are activated by specific serine proteases, which are expressed in many normal and malignant cell types. Previous in vitro studies on prostate cancer cells suggest a role for PAR-2 in prostate cancer metastasis. A polyclonal anti-human TMPRSS2 antibody was generated against the TMPRSS2 serine protease domain. The antibody showed specific reactivity with recombinant expressed TMPRSS2, and so was used to extract and purify the cleaved active TMPRSS2 protease from prostate cancer cells. Reverse transcriptase PCR and Western blot analysis were used to show the expression of both TMPRSS2 and PAR-2 in the androgen-dependent LNCaP prostate cancer cell line. Treatment of LNCaP cells with the cellular immunopurified TMPRSS2 protease induced a transient increase in intracellular calcium, which is indicative of G-protein-coupled-receptor activation. This calcium mobilization was inhibited by cellular pre-treatment with a specific PAR-2 antagonist, but not with a PAR-1 antagonist; inhibition of the protease activity also failed to mobilize calcium, suggesting that TMPRSS2 is capable of cleaving and thereby activating the PAR-2 receptor. The calcium mobilization was also inhibited by cellular pre-treatment with suramin or 2-APB (2-aminoethoxydiphenyl borate), indicating that a G-protein pathway is involved and that subsequent calcium release is mainly from intracellular stores. The present study describes how TMPRSS2 may contribute to prostate tumour metastasis via the activation of PAR-2.
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Research Article| June 07 2005
The membrane-anchored serine protease, TMPRSS2, activates PAR-2 in prostate cancer cells
Susan WILSON;
Susan WILSON
*School of Pharmacy, Medical Biology Centre, Queen's University Belfast, 97 Lisburn Road, Belfast BT9 7BL, Northern Ireland, U.K.
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Brett GREER;
Brett GREER
†School of Biology and Biochemistry, Medical Biology Centre, Queen's University Belfast, 97 Lisburn Road, Belfast BT9 7BL, Northern Ireland, U.K.
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John HOOPER;
John HOOPER
‡Molecular and Cell Biology, School of Life Sciences, Queensland University of Technology, Brisbane, Australia
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Andries ZIJLSTRA;
Andries ZIJLSTRA
§Department of Cell Biology, The Scripps Research Institute, La Jolla, CA 92093, U.S.A.
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Brian WALKER;
Brian WALKER
*School of Pharmacy, Medical Biology Centre, Queen's University Belfast, 97 Lisburn Road, Belfast BT9 7BL, Northern Ireland, U.K.
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James QUIGLEY;
James QUIGLEY
§Department of Cell Biology, The Scripps Research Institute, La Jolla, CA 92093, U.S.A.
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Susan HAWTHORNE
Susan HAWTHORNE 1
*School of Pharmacy, Medical Biology Centre, Queen's University Belfast, 97 Lisburn Road, Belfast BT9 7BL, Northern Ireland, U.K.
1To whom correspondence should be addressed (email s.hawthorne@qub.ac.uk).
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Publisher: Portland Press Ltd
Received: June 23 2004
Revision Received: November 03 2004
Accepted: November 10 2004
Accepted Manuscript online: November 10 2004
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London
2005
Biochem J (2005) 388 (3): 967–972.
Article history
Received:
June 23 2004
Revision Received:
November 03 2004
Accepted:
November 10 2004
Accepted Manuscript online:
November 10 2004
Citation
Susan WILSON, Brett GREER, John HOOPER, Andries ZIJLSTRA, Brian WALKER, James QUIGLEY, Susan HAWTHORNE; The membrane-anchored serine protease, TMPRSS2, activates PAR-2 in prostate cancer cells. Biochem J 15 June 2005; 388 (3): 967–972. doi: https://doi.org/10.1042/BJ20041066
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