Thrombin conducts epithelial‑mesenchymal transition via protease‑activated receptor‑1 in human gastric cancer

Otsuki,Tadayoshi; Fujimoto,Daisuke; Hirono,Yasuo; Goi,Takanori; Yamaguchi,Akio

doi:10.3892/ijo.2014.2651

Thrombin conducts epithelial‑mesenchymal transition via protease‑activated receptor‑1 in human gastric cancer

Authors:
- Tadayoshi Otsuki
- Daisuke Fujimoto
- Yasuo Hirono
- Takanori Goi
- Akio Yamaguchi
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Affiliations: First Department of Surgery, Faculty of Medicine, University of Fukui, Fukui 910‑1193, Japan
Published online on: September 15, 2014 https://doi.org/10.3892/ijo.2014.2651
Pages: 2287-2294

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Abstract

Epithelial‑mesenchymal transition (EMT) is thought to be a key step for cancer metastasis. Using an immunohistochemical approach with gastric carcinoma tissue, we found the expression of protease‑activated receptor‑1 (PAR1), along with a metalloproteinase known to activate PAR1, were associated with poorer prognosis, compared with expression‑negative tumors, and activated PAR1 promotes gastric cancer cell invasion and proliferation in vivo. In this study we observed EMT induction by the PAR1 agonist α‑thrombin, in human gastric cell lines stably expressing PAR1. We investigated α‑thrombin-induced changes in the cell forms of pcDNA3.1‑MKN45 (MKN45/Mock), pcDNA3.1‑PAR1 transfected MKN45 (MKN45/PAR1), and MKN74. Expression levels of epithelial and mesenchymal markers as well as the distribution of transcriptional factors of E‑cadherin in the cytoplasm and nucleus were also noted in these cell lines. We observed α‑thrombin-induced morphological changes in MKN45/PAR1 and MKN74 cells. Western blotting and immunohistochemistry of these cells indicated a fall in the expression level of E‑cadherin and an increase in fibronectin expression after 48 h. PAR1 activation also induced significant increases in nuclear levels of the Snail which is a repressor of E‑cadherin gene expression. We found EMT in gastric cancer cell lines that underwent α‑thrombin-induced PAR1 activation.

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