miRNA expression microarrays in GM-CSF or GM-CSF+IL6-induced mouse MDSC. miRNA expression microarrays in GM-CSF or GM-CSF+IL6-induced mouse MDSC

Myeloid-derived suppressor cells (MDSCs) have emerged as major regulators of immune responses in cancer and other pathological conditions. Multiple factors including cytokines, transcription factors and multiple signaling pathways are involved in MDSC differentiation. Cytokines such as granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin(IL-6) etc could in vitro mediate development of MDSCs.IL-6 with GM-CSF mediated MDSC not only had stronger suppressive function but also the dynamics of their suppressive function was different from GM-CSF alone mediated MDSCs.To found a new regulatory factor (s) in tumor and inflammatory environments, we compared GM-CSF and IL-6 mediated MDSCs with GM-CSF alone mediated MDSCs using lncRNA microarray, miRNA microarrays and protein-coding mRNA microarrays. Overall design: Inflammatory factors associated MDSCs were generated by cultivating bone marrow cells of C57BL/6 mice in petri dishes for 4 days in the presence of 5% FBS medium containing GM-CSF (40 ng/ml) only, GM-CSF (40 ng/ml) plus IL-6 (40 ng/ml) using different processing group for each experiment.

髓系来源抑制细胞（Myeloid-derived suppressor cells，MDSCs）现已被证实为癌症及其他病理状态下免疫应答的核心调控因子。该类细胞的分化过程涉及多种调控因素，包括细胞因子、转录因子及多条信号通路。诸如粒细胞-巨噬细胞集落刺激因子（GM-CSF）、白细胞介素-6（IL-6）等细胞因子可在体外介导髓系来源抑制细胞的发育。相较于仅经GM-CSF诱导的髓系来源抑制细胞，同时经GM-CSF与IL-6诱导的髓系来源抑制细胞不仅具备更强的免疫抑制功能，其免疫抑制功能的动态变化特征也存在显著差异。为筛选肿瘤与炎症微环境中的新型调控因子，本研究通过长链非编码RNA（lncRNA）微阵列、微小RNA（miRNA）微阵列及编码蛋白mRNA微阵列，对比了仅由GM-CSF诱导的髓系来源抑制细胞与GM-CSF联合IL-6诱导的髓系来源抑制细胞的表达谱差异。实验整体设计：将C57BL/6小鼠的骨髓细胞接种于培养皿中，在含5%胎牛血清（FBS）的培养基内培养4天，分别采用仅添加40 ng/ml GM-CSF、以及同时添加40 ng/ml GM-CSF与40 ng/ml IL-6的两种处理方式，诱导生成炎症相关髓系来源抑制细胞；每个实验均设置独立处理组。