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Protease-activated-receptor-2 affects protease-activated-receptor-1-driven breast cancer

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An Erratum to this article was published on 30 November 2016

Abstract

Mammalian protease-activated-receptor-1 and -2 (PAR1 and PAR2) are activated by proteases found in the flexible microenvironment of a tumor and play a central role in breast cancer. We propose in the present study that PAR1 and PAR2 act together as a functional unit during malignant and physiological invasion processes. This notion is supported by assessing pro-tumor functions in the presence of short hairpin; shRNA knocked-down hPar2 or by the use of a truncated PAR2 devoid of the entire cytoplasmic tail. Silencing of hPar2 by shRNA-attenuated thrombin induced PAR1 signaling as recapitulated by inhibiting the assembly of Etk/Bmx or Akt onto PAR1-C-tail, by thrombin-instigated colony formation and invasion. Strikingly, shRNA-hPar2 also inhibited the TFLLRN selective PAR1 pro-tumor functions. In addition, while evaluating the physiological invasion process of placenta extravillous trophoblast (EVT) organ culture, we observed inhibition of both thrombin or the selective PAR1 ligand; TFLLRNPNDK induced EVT invasion by shRNA-hPar2 but not by scrambled shRNA-hPar2. In parallel, when a truncated PAR2 was utilized in a xenograft mouse model, it inhibited PAR1–PAR2-driven tumor growth in vivo. Similarly, it also attenuated the interaction of Etk/Bmx with the PAR1-C-tail in vitro and decreased markedly selective PAR1-induced Matrigel invasion. Confocal images demonstrated co-localization of PAR1 and PAR2 in HEK293T cells over-expressing YFP-hPar2 and HA-hPar1. Co-immuno-precipitation analyses revealed PAR1-PAR2 complex formation but no PAR1-CXCR4 complex was formed. Taken together, our observations show that PAR1 and PAR2 act as a functional unit in tumor development and placenta-uterus interactions. This conclusion may have significant consequences on future breast cancer therapeutic modalities and improved late pregnancy outcome.

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Acknowledgments

We are grateful to Dr. Susan Lewis for editing the manuscript. This work was supported by grants from the Israel Science Foundation, and Nofar Chief Israeli Scientist, Trade and Industrial Office (to Bar-Shavit, R).

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There are no potential conflicts of interest to disclose.

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Authors and Affiliations

  1. Sharett-Institute of Oncology, Hadassah-Hebrew University Medical Center, POB 12000, 91120, Jerusalem, Israel

    Mohammad Jaber, Miriam Maoz, Arun Kancharla, Daniel Agranovich, Tamar Peretz, Beatrice Uziely & Rachel Bar-Shavit

  2. Department of Obstetrics and Gynecology, Shaare-Zedek and Hadassah-Hebrew University Medical Centers, POB 12000, 91120, Jerusalem, Israel

    Sorina Grisaru-Granovsky

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  1. Mohammad Jaber
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  2. Miriam Maoz
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  3. Arun Kancharla
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Correspondence to Rachel Bar-Shavit.

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An erratum to this article is available at http://dx.doi.org/10.1007/s00018-016-2424-6.

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Jaber, M., Maoz, M., Kancharla, A. et al. Protease-activated-receptor-2 affects protease-activated-receptor-1-driven breast cancer. Cell. Mol. Life Sci. 71, 2517–2533 (2014). https://doi.org/10.1007/s00018-013-1498-7

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