Cathepsin K induces platelet dysfunction and affects cell signaling in breast cancer - molecularly distinct behavior of cathepsin K in breast cancer

Author(s):Show less -

Andrade, Sheila Siqueira ^{[1, 2, 3]} ; Gouvea, Iuri Estrada ^[4] ; Silva, Mariana Cristina C. ^[5] ; Castro, Eloisa Dognani ^[5] ; de Paula, Claudia A. A. ^[5] ; Okamoto, Debora ^[4] ; Oliveira, Lilian ^[4] ; Peres, Giovani Bravin ^[5] ; Ottaiano, Tatiana ^[5] ; Facina, Gil ^{[1, 2]} ; Pinto Nazario, Afonso Celso ^{[1, 2]} ; Campos, Antonio Hugo J. F. M. ^[6] ; Paredes-Gamero, Edgar Julian ^[5] ; Juliano, Maria ^[4] ; da Silva, Ismael D. C. G. ^{[1, 2]} ; Oliva, Maria Luiza V. ^[5] ; Girao, Manoel J. B. C. ^{[1, 2]}

Affiliation

^[1] Univ Fed Sao Paulo, Dept Gynecol, BR-04024002 Sao Paulo, SP - Brazil

^[2] COLSAN, Charitable Assoc Blood Collect, BR-04080006 Sao Paulo, SP - Brazil

^[3] Univ Fed Sao Paulo, Cellular Gynecol Lab, Dept Gynecol, Rua Napoleao Barros 608, BR-04024002 Sao Paulo - Brazil

^[4] Univ Fed Sao Paulo, Dept Biophys, BR-04024002 Sao Paulo, SP - Brazil

^[5] Univ Fed Sao Paulo, Dept Biochem, BR-04024002 Sao Paulo, SP - Brazil

^[6] Antonio Prudente Fdn, AC Camargo Canc Ctr, AC Camargo Hosp Biobank, Dept Pathol, BR-01509010 Sao Paulo, SP - Brazil

• Document type: Journal article
• Source: BMC CANCER; v. 16, MAR 1 2016.
• Web of Science Citations: 7

Abstract

Background: Breast cancer comprises clinically and molecularly distinct tumor subgroups that differ in cell histology and biology and show divergent clinical phenotypes that impede phase III trials, such as those utilizing cathepsin K inhibitors. Here we correlate the epithelial-mesenchymal-like transition breast cancer cells and cathepsin K secretion with activation and aggregation of platelets. Cathepsin K is up-regulated in cancer cells that proteolyze extracellular matrix and contributes to invasiveness. Although proteolytically activated receptors (PARs) are activated by proteases, the direct interaction of cysteine cathepsins with PARs is poorly understood. In human platelets, PAR-1 and -4 are highly expressed, but PAR-3 shows low expression and unclear functions. Methods: Platelet aggregation was monitored by measuring changes in turbidity. Platelets were immunoblotted with anti-phospho and total p38, Src-Tyr-416, FAK-Tyr-397, and TGF beta monoclonal antibody. Activation was measured in a flow cytometer and calcium mobilization in a confocal microscope. Mammary epithelial cells were prepared from the primary breast cancer samples of 15 women with Luminal-B subtype to produce primary cells. Results: We demonstrate that platelets are aggregated by cathepsin K in a dose-dependent manner, but not by other cysteine cathepsins. PARs-3 and -4 were confirmed as the cathepsin K target by immunodetection and specific antagonists using a fibroblast cell line derived from PARs deficient mice. Moreover, through co-culture experiments, we show that platelets activated by cathepsin K mediated the up-regulation of SHH, PTHrP, OPN, and TGF beta in epithelial-mesenchymal-like cells from patients with Luminal B breast cancer. Conclusions: Cathepsin K induces platelet dysfunction and affects signaling in breast cancer cells. (AU)

• FAPESP's process: 12/19851-8 - Heterotypic culture: The Cross-Talking between epithelial tumor cells, mammary stroma, macrophages and platelets in breast cancer: the release of proteolytic enzymes and signaling of Toll-like receptors
• Grantee: Sheila Siqueira Andrade
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 09/53766-5 - Proteins from plant source with selectivity for inhibition of mammalian enzymes and their role as an anti-inflammatory, antithrombotic, anti-diabetic and anti-tumor agent
• Grantee: Maria Luiza Vilela Oliva
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 12/19780-3 - Heterotypic culture: the cross-talking between epithelial tumor cells, mammary stroma, macrophages and platelets in breast cancer: the release of proteolytic enzymes and signaling of toll-like receptors
• Grantee: Manoel João Batista Castello Girão
• Support Opportunities: Regular Research Grants