Effects of EPCR on the proliferation and migration of human breast cancer cells MDA-MB-231

XU Yanyan; ZHUO Qian; TANG Yangyang; WANG Qingling

doi:10.3969/j.issn.1002-266X.2018.09.004

Abstract：Objective To explore the effects and mechanism of endothelial protein C receptor (EPCR) on the proliferation and migration of human breast cancer cells MDA-MB-231.Methods MDA-MB-231 cells were divided into the EPCR transfection group,unrelated sequence transfection group,and control group which were transfected with EPCR-siRNA,siRNA-NC,and empty plasmid,respectively.Cell-ELISA was used to detect the activation of PAR-1 in the membranes of MDA-MB-231.CCK-8 assay wasperformed to observe the proliferation of cells in each group.Transwell chamber was employed to observe the cell's migration.After treatment of anti-PAR-1 antibody,the proliferation and migration abilities of MDA-MB-23 1 cells were detected by CCK-8 and Transwell assay.Western blotting was used to determine the protein expression levels of Erk1/2 and AKT.Results Compared with the control group and unrelated sequence transfection group,the binding rate of un-cleaved activated PAR-1 antibody increased,and the proliferation and migration abilities significantly decreased in the transfection group (all P ＜ 0.05).Compared with the control group,after being treated with anti-PAR-1 antibody,the proliferation and migration of MDA-MB-231 cells significantly decreased (both P ＜0.05).After knockdown of EPCR or the PAR-1 antibody treatment,the expression of p-Akt (S473) and p-Akt (T308) significantly decreased in the MDA-MB-231 cells (both P ＜ 0.05),but the expression of p-Erk1/2 did not change significantly (P ＞0.05).Conclusion EPCR contributes to the proliferation and migration of MDA-MB-231 cells by activating AKT,and this effect of EPCR may be dependent on PAR-1.