title: Activation of platelet protease-activated receptor-1 induces epithelial-mesenchymal transition and chemotaxis of colon cancer cell line SW620.
abstract: The aim of the present study was to examine the role of protease-activated receptor-1 (PAR1)-stimulated platelet activation in the epithelial-mesenchymal transition (EMT) and migration of colon cancer cells, and to identify the underlying mechanisms. TFLLR-NH2, a PAR1 agonist, was used to activate platelets and the platelet supernatants were used to treat the SW620 colon cancer cell line. Expression of E-cadherin and vimentin on SW620 cells was detected by immunofluorescence and western blotting, and the level of the transforming growth factor β1 (TGF-β1) was measured using ELISA following the activation of platelets by TFLLR-NH2. miR-200b expression was detected using quantitative PCR in SW620 cells. In order to investigate the chemotactic ability of the SW620 cells, the expression of CXC chemokine receptor type 4 (CXCR4) was measured by flow cytometry. Transwell migration assays were performed following exposure of the cells to the supernatant of PAR1-activated platelets. SW620 cells cultured in the supernatant of TFLLR-NH2-activated platelets upregulated E-cadherin expression and downregulated the vimentin expression. In the in vitro platelet culture system, a TFLLR-NH2 dose-dependent increase of secreted TGF-β1 was detected in the supernatant. The activation of PAR1 on the platelets led to the inhibition of miR-200b expression in the SW620 cells that were cultured in platelet-conditioned media. The number of SW620 cells that penetrated through the Transwell membrane increased with the dose of TFLLR-NH2 used to treat the platelets. The percentage of CXCR4-positive SW620 cells was significantly higher when they were exposed to the supernatant of platelets cultured for 24 h with PAR1 agonist than when cultured in non-conditioned media (40.89 ± 6.74 vs. 3.47 ± 1.40%, P < 0.01). Platelet activation with a PAR1 agonist triggered TGF-β secretion, which induced EMT of SW620 human colon cancer cells via the downregulation of miR-200b expression, and activated platelets had a chemotactic effect on colon cancer cells mediated by the upregulation of CXCR4 on the cell surface.
authors.affiliation: Department of Oncology, Hebei General Hospital, Shijiazhuang, Hebei 050051, P.R. China.
authors.forename: Yitao
authors.initials: Y
authors.lastname: Jia
authors.affiliation: Department of Oncology, Hebei General Hospital, Shijiazhuang, Hebei 050051, P.R. China.
authors.forename: Suqiao
authors.initials: S
authors.lastname: Zhang
authors.affiliation: Second Department of Surgery, The Fourth Hospital of Hebei Medical University, Shijiazhuang, Hebei 050011, P.R. China.
authors.forename: Lingling
authors.initials: L
authors.lastname: Miao
authors.affiliation: Second Department of Surgery, The Fourth Hospital of Hebei Medical University, Shijiazhuang, Hebei 050011, P.R. China.
authors.forename: Jingbao
authors.initials: J
authors.lastname: Wang
authors.affiliation: Second Department of Surgery, The Fourth Hospital of Hebei Medical University, Shijiazhuang, Hebei 050011, P.R. China.
authors.forename: Zujian
authors.initials: Z
authors.lastname: Jin
authors.affiliation: Second Department of Surgery, The Fourth Hospital of Hebei Medical University, Shijiazhuang, Hebei 050011, P.R. China.
authors.forename: Bin
authors.initials: B
authors.lastname: Gu
authors.affiliation: Second Department of Surgery, The Fourth Hospital of Hebei Medical University, Shijiazhuang, Hebei 050011, P.R. China.
authors.forename: Zhihui
authors.initials: Z
authors.lastname: Duan
authors.affiliation: Second Department of Surgery, The Fourth Hospital of Hebei Medical University, Shijiazhuang, Hebei 050011, P.R. China.
authors.forename: Zhaolong
authors.initials: Z
authors.lastname: Zhao
authors.affiliation: Department of Surgery, Hebei Medical University Affiliated North China Petroleum Bureau General Hospital, Renqiu, Hebei 062552, P.R. China.
authors.forename: Shunmao
authors.initials: S
authors.lastname: Ma
authors.affiliation: Centre of Breast Cancer, The Fourth Hospital of Hebei Medical University, Shijiazhuang, Hebei 050011, P.R. China.
authors.forename: Wenjin
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Second Department of Surgery, The Fourth Hospital of Hebei Medical University, Shijiazhuang, Hebei 050011, P.R. China.
authors.forename: Zhongxin
authors.initials: Z
authors.lastname: Li
mesh_terms: D015820:Cadherins / Q000096:biosynthesis; D045744:Cell Line, Tumor; D002465:Cell Movement; D002633:Chemotaxis / Q000187:drug effects; D003110:Colonic Neoplasms / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics; D006801:Humans; D035683:MicroRNAs / Q000096:biosynthesis / Q000235:genetics*; D009842:Oligopeptides / Q000008:administration & dosage; D015539:Platelet Activation / Q000235:genetics; D044463:Receptor, PAR-1 / Q000096:biosynthesis* / Q000235:genetics; D019718:Receptors, CXCR4 / Q000096:biosynthesis / Q000235:genetics*; D053773:Transforming Growth Factor beta1 / Q000096:biosynthesis / Q000235:genetics*; D014746:Vimentin / Q000096:biosynthesis
chemical_list: C527276:CXCR4 protein, human; D015820:Cadherins; C533896:MIRN200 microRNA, human; D035683:MicroRNAs; D009842:Oligopeptides; C462999:PAR-1-activating peptide; D044463:Receptor, PAR-1; D019718:Receptors, CXCR4; D053773:Transforming Growth Factor beta1; D014746:Vimentin
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