| |
| Purpose: To investigate the e!ect of N-glycan-defective mammary adenocarcinoma cells
on the polarization of macrophages. Methods: N-glycan-defective breast cancer cells (MA782 cells) were prepared by swainsonine(SW) treatment and the cytotoxicity of SW to MA782 cells was evaluated using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. "e Nglycan-defective MA782 cells were co-cultured with bone marrow-derived macrophages (BMDMs) for 48 h in vitro, and then the BMDMs and the co-cultured supernatant were analyzed for macrophage phenotypic using FQRT-PCR, FCM and ELISA. Results: SW-treated MA782 cells expressed defective N-glycan on the cell surface in a dose-dependent manner (*p<0.05). MTT assays showed that neither the 1 ug/mL nor 5ug/ mL SW treatments showed signi$cant inhibition of MA782 cell growth in vitro. "e expression of iNOS and agr-1 in the 5 #g/mL SW-treated group were 4.75-fold higher and 3.7-fold lower than that in the untreated group, respectively (*p<0.05). Mean %uorescence intensity of CD16/32 expressed in the cells treated with 5 #g/mL SW was signi$cantly higher in comparison with the untreated group (65 vs. 7, *p<0.05), though the percentage of CD16/32-positive cells were not signi$cantly di!erent. Furthermore, the expression of CD206 and dectin-1 in the 5 #g/mL SW-treated group was signi$cantly decreased (3.1±0.3% and 4.1±1.1%, respectively) in comparison with the untreated group (40±3% and 8.9±1.2%, respectively, both p<0.05). In addition, the 5 #g/mL SW-treated group secreted more TNF-alpha (350 ±25 pg/mL) and less IL-10 (89±7.2 pg/mL) than the untreated group (80 ±3 pg/mL and 150 ±10 pg/mL, respectively, both p<0.05). Conclusion: N-glycan-defective MA782 cells can induce the di!erentiation of BMDM
into proin%ammatory M1 macrophages in vitro. |
|
| Keywords:macrophages; breast cancer cells; swainsonine; phenotype |
|
