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

BMC Cancer

Research Article

Mariana Cristina C. Silva¹  Giovani Bravin Peres¹  Eloísa Dognani Castro¹  Maria Luiza V. Oliva¹  Tatiana Ottaiano¹  Edgar Julian Paredes-Gamero¹  Cláudia A. A. de Paula¹  Debora Okamoto²  Iuri Estrada Gouvea²  Lilian Oliveira²  Maria Juliano²  Antonio Hugo J. F. M. Campos³  Ismael D. C. G. da Silva⁴  Afonso Celso Pinto Nazário⁴  Gil Facina⁴  Manoel J. B. C. Girão⁵  Sheila Siqueira Andrade⁶ 

[1] Biochemistry of Universidade Federal de São Paulo, 04024-002, São Paulo, SP, Brazil;Biophysics of Universidade Federal de São Paulo, 04024-002, São Paulo, SP, Brazil;Department of Pathology, AC Camargo Hospital Biobank, A C Camargo Cancer Center - Antonio Prudente Foundation, 01509-010, São Paulo, SP, Brazil;Departments of Gynecology of Universidade Federal de São Paulo, 04024-002, São Paulo, SP, Brazil;Departments of Gynecology of Universidade Federal de São Paulo, 04024-002, São Paulo, SP, Brazil;Charitable Association of Blood Collection – COLSAN, 04080-006, São Paulo, SP, Brazil;Departments of Gynecology of Universidade Federal de São Paulo, 04024-002, São Paulo, SP, Brazil;Charitable Association of Blood Collection – COLSAN, 04080-006, São Paulo, SP, Brazil;Department of Gynecology, Cellular Gynecology Laboratory, Universidade Federal de São Paulo, Rua Napoleão de Barros, 608, CEP 04024-002, São Paulo, Brazil;

关键词: Cathepsin K;    Platelets;    Breast cancer;    Protease activated receptors;   

DOI  :  10.1186/s12885-016-2203-7

 received in 2015-09-30, accepted in 2016-02-17,  发布年份 2016

来源: Springer

【 摘 要 】

BackgroundBreast 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.MethodsPlatelet aggregation was monitored by measuring changes in turbidity. Platelets were immunoblotted with anti-phospho and total p38, Src-Tyr-416, FAK-Tyr-397, and TGFβ 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.ResultsWe 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 TGFβ in epithelial-mesenchymal-like cells from patients with Luminal B breast cancer.ConclusionsCathepsin K induces platelet dysfunction and affects signaling in breast cancer cells.

【 授权许可】

CC BY   
© Andrade et al. 2016