title: Thrombin conducts epithelial‑mesenchymal transition via protease‑activated receptor‑1 in human gastric cancer.
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.
authors.affiliation: First Department of Surgery, Faculty of Medicine, University of Fukui, Fukui 910‑1193, Japan.
authors.forename: Tadayoshi
authors.initials: T
authors.lastname: Otsuki
authors.affiliation: First Department of Surgery, Faculty of Medicine, University of Fukui, Fukui 910‑1193, Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Fujimoto
authors.affiliation: First Department of Surgery, Faculty of Medicine, University of Fukui, Fukui 910‑1193, Japan.
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Hirono
authors.affiliation: First Department of Surgery, Faculty of Medicine, University of Fukui, Fukui 910‑1193, Japan.
authors.forename: Takanori
authors.initials: T
authors.lastname: Goi
authors.affiliation: First Department of Surgery, Faculty of Medicine, University of Fukui, Fukui 910‑1193, Japan.
authors.forename: Akio
authors.initials: A
authors.lastname: Yamaguchi
mesh_terms: D015820:Cadherins / Q000096:biosynthesis; D045744:Cell Line, Tumor; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008297:Male; D009361:Neoplasm Invasiveness / Q000235:genetics; D009362:Neoplasm Metastasis; D044463:Receptor, PAR-1 / Q000096:biosynthesis* / Q000235:genetics; D013274:Stomach Neoplasms / Q000235:genetics* / Q000473:pathology; D013917:Thrombin / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D015820:Cadherins; D044463:Receptor, PAR-1; D013917:Thrombin