来那度胺可抑制单核细胞向M2型巨噬细胞分化及相关细胞因子的分泌

Lenalidomide significantly inhibits the differentiation of monocytes to M2 polarized macrophages and down-regulates secretion of associated cytokines

目的探索来那度胺在肿瘤微环境中对于单核细胞向M2型巨噬细胞分化及IL-10、VEGF和TGF-β1水平的影响。方法密度梯度离心法分离初治淋巴瘤患者及健康志愿者外周血单个核细胞(PBMCs)。用Transwell24孔板共培养PBMCs与淋巴瘤细胞系HUT-78,并于共培养体系中加入来那度胺。流式细胞术分析肿瘤相关巨噬细胞(TAM)特征性表型CD68和CD163的表达,ELISA法检测共培养体系细胞因子IL-10、VEGF和TGF-β1的含量。结果在PBMCs与HUT-78构成的体外共培养体系中加入来那度胺后,CD68+、CD163+、CD68+CD163+细胞及CD68+CD163+在CD68+细胞中所占比例均有显著的下降(P<0.05)。加入来那度胺后患者组CD68+CD163+/CD68+下降程度明显高于健康志愿者(P<0.05)。加入来那度胺的患者共培养组相对于未加来那度胺的共培养组IL-10和VEGF水平均有显著下降(P<0.01,P<0.05)。结论来那度胺在PBMCs与淋巴瘤细胞构建的体外共培养体系中能够有效抑制单核细胞向M2型TAM的分化,且能够抑制患者组共培养体系中IL-10、VEGF分泌。

Objective To explore the impact of anti-cancer drug Lenalidomide on the differentiation of monocytes to M2 polarized macrophages and the secretion of IL-10,VEGF and TGF-β1.Methods The peripheral blood mononuclear cells （PBMCs） were isolated by Ficoll-Hypaque density gradient centrifugation from peripheral blood of both lymphoma patients and healthy controls.Then PBMCs were co-cultured with HUT-78 lymphoma cells in Transwell 24-well plates.Anti-cancer drug Lenalidomide was added into the co-culture system.Expressions of the markers of TAMs （CD68 and CD163） were tested by flow cytometry.IL-10,VEGF and TGF-β1 level in the co-culture system were detected by ELISA.Results Treatment with Lenalidomide significantly decreased the proportion of CD68＋,CD163 ＋,CD68 ＋ CD163 ＋ cells and the proportion of CD68 ＋ CD163 ＋ / CD68 ＋ cells（P ＜ 0.05）.The degree of decline of CD68 ＋ CD163 ＋/CD68 ＋ cells was more significant in patients than that in healthy controls（P ＜ 0.05）.The reatment with Lenalidomide significantly decreased the IL-10 and VEGF level in the co-culture system （P ＜ 0.01,P ＜ 0.05）.Conclusions Lenalidomide significantly inhibits the differentiation of monocytes to M2 polarized macrophages and down-regulate the secretion of IL-10 and VEGF in the co-culture system in patient group.