Objective To investigate the effect of M1 macrophages on epithelial-mesenchymal transition(EMT)of breast cancer cells and the mediated role of Calpain.Methods Estrogen receptor(ER+)positive human breast cancer cells MCF-7,T47D,and ER-negative human breast cancer cells MDA-MD-231 were used as model cells.In the experiment,MCF-7 and T47D cells were divided into RPMI-1640 group(RPMI-1640 pure medium culture),M1-CM group(M1 macrophage conditioned medium culture group),M1-CM+Calp group[M1-CM combined with Calpain inhibitor Calpeptin(50 μmoL/L)],and M1-CM+CI-Ⅲ group[M1-CM combined with Calpain inhibitor Calpain inhibitor Ⅲ(50 µmoL/L)];MDA-MB-231 cells were divided into RPMI-1640 group and M1-CM group.The cell migration ability of MCF-7 cells,T47D cells,and MDA-MB-231 cells and the cell invasion ability of MCF-7 cells in each group were detected by Wound-healing assay and Transwell assay.The mRNA expression levels of CC-chemokine receptor 7(CCR7),CXC motif chemokine ligand 10(CXCL10)and interleukin-12(IL-12)in M0 macrophages and M1 macrophages were detected by qRT-PCR.The protein levels of epithelial cadherin(E-cad),fibritin(FN),and vimentin(Vim)in MCF-7 cells,T47D cells,and MDA-MB-231 cells were detected by Western blot.Results Compared with the RPMI-1640 group,the migration ability of ER+cells MCF-7 and T47D in the M1-CM group was enhanced(P＜0.05),and the invasive ability of MCF-7 cells was enhanced(P＜0.05).The expression levels of MCF-7 and E-cad in ER+cells decreased,while the expressions of FN and Vim increased(P＜0.05),and only E-cad expression increased in ER-cells MDA-MB-231(P＜0.05).Compared with the M1-CM group,the EMT effect of ER+breast cancer cells induced by M1-CM in the M1-CM+Calp group and M1-CM+CI-Ⅲ group of ER positive cells MCF-7 and T47D was reversed(P＜0.05).Conclusion M1 macrophages conditioned medium can promote EMT of ER+cells and human breast cells,but has no obvious effect on EMT of ER-cells.Its mechanism of promoting EMT of ER+breast cancer cells might be related to Calpain signaling.
breast cancerM1 macrophagescell invasionepithelial-mesenchymal transitioncalpainestrogen receptor
维普
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