title: N-glycan-defective breast cancer cells induce a phenotypic switch in polarization of bone marrow-derived macrophages.
abstract: OBJECTIVE
  To investigate the effect 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. The N-glycan-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 μg/mL nor 5 μg/mL SW treatments showed significant inhibition of MA782 cell growth in vitro. The expression of iNOS and agr-1 in the 5 μg/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 fluorescence intensity of CD16/32 expressed in the cells treated with 5 μg/mL SW was significantly higher in comparison with the untreated group (65 vs. 7, *p < 0.05), though the percentage of CD16/32-positive cells were not significantly different. Furthermore, the expression of CD206 and dectin-1 in the 5 μg/mL SW-treated group was significantly 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 μg/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).
  
  
  CONCLUSIONS
  N-glycan-defective MA782 cells can induce the differentiation of BMDM into proinflammatory M1 macrophages in vitro.
authors.affiliation: Department of Immunology, School of Basic Medical Science, Wuhan University, Wuhan, Hubei, P.R. China.
authors.forename: Haidan
authors.initials: H
authors.lastname: Chen
authors.forename: Huili
authors.initials: H
authors.lastname: Cai
authors.forename: Liang
authors.initials: L
authors.lastname: Chen
authors.forename: Xianglei
authors.initials: X
authors.lastname: Wu
authors.forename: Dongqing
authors.initials: D
authors.lastname: Li
mesh_terms: D001483:Base Sequence; D001854:Bone Marrow Cells / Q000166:cytology*; D001943:Breast Neoplasms / Q000378:metabolism / Q000473:pathology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D017931:DNA Primers; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D008264:Macrophages / Q000166:cytology*; D010641:Phenotype; D011134:Polysaccharides / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017026:Swainsonine / Q000494:pharmacology
chemical_list: D017931:DNA Primers; D011134:Polysaccharides; D017026:Swainsonine