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目的探讨肿瘤抗原MUC1抑制骨髓来源抑制细胞(myeloid-derived suppressor cell,MDSC)产生的机制。方法分离C57BL/6小鼠股骨骨髓细胞,分别与稳定表达含22个串联重复序列的全长人MUC1 cDNA片段的B16细胞(B16-MUC1)和空质粒对照B16细胞(B16-neo)培养上清共培养,流式细胞术分析MUC1对MDSC产生及MDSC表达MUC1蛋白的影响;瑞氏姬姆萨染色观察MUC1对骨髓细胞生长形态的影响;流式细胞术检测共培养的骨髓细胞中单核细胞和巨噬细胞标志物CD14和F4/80的表达及成熟单核-巨噬细胞标志物CD11b和CD68的表达。结果正常C57BL/6小鼠骨髓细胞分别与B16-neo和B16-MUC1细胞培养上清共培养24和48 h后,BM+B16-MUC1组骨髓细胞中CD11b+Gr-1+MDSC数量较BM+B16-neo组均明显减少(P<0.05),共培养48 h后,BM+B16-MUC1组的CD11b+Gr-1+MDSC中MUC1蛋白的表达较BM+B16-neo组明显增多(P<0.01);BM+B16-MUC1组骨髓细胞中单核-巨噬细胞数量增多;与BM+B16-neo组相比,BM+B16-MUC1组CD14+、F4/80+和CD68+细胞表达数量明显增高(分别为P<0.01、P<0.01、P<0.05),CD11b+平均荧光强度明显增高(P<0.01)。结论 MUC1通过诱导骨髓细胞向单核-巨噬细胞分化,从而抑制MDSC的产生。
Objective To investigate the mechanism of tumor antigen MUC1 inhibiting myeloid-derived suppressor cell (MDSC). Methods C57BL / 6 mouse femur bone marrow cells were isolated and compared with B16-MUC1 cells stably expressing the full-length human MUC1 cDNA fragment containing 22 tandem repeats (B16-MUC1) and B16-neo culture supernatants The effects of MUC1 on the production of MDSCs and the expression of MUC1 protein in MDSCs were analyzed by flow cytometry. The effects of MUC1 on the morphology of bone marrow cells were observed by Wright Giesa staining. The expression of MUC1 in mononuclear cells And the expression of macrophage markers CD14 and F4 / 80 and the expression of mature monocyte-macrophage markers CD11b and CD68. Results The number of CD11b + Gr-1 + MDSC in BM + B16-MUC1 group was significantly higher than that in BM + B16-MUC1 group after C57BL / 6 mouse bone marrow cells were cocultured with B16-neo and B16- (P <0.05). After co-cultured for 48 h, the expression of MUC1 in CD11b + Gr-1 + MDSC of BM + B16-MUC1 group was significantly higher than that in BM + B16-neo group (P < 0.01). The number of monocyte-macrophage cells in BM + B16-MUC1 group increased compared with BM + B16-neo group, and the numbers of CD14 +, F4 / 80 + and CD68 + cells in BM + B16- (P <0.01, P <0.01, P <0.05, respectively). The average fluorescence intensity of CD11b + was significantly increased (P <0.01). Conclusion MUC1 can inhibit the production of MDSCs by inducing bone marrow cells to differentiate into monocytes and macrophages.