不同分化阶段树突状细胞肌动蛋白微丝结合蛋白的表达变化

The Changes of Expression Levels of F-actin Binding Proteins in Dendritic Cells at Different Differentiation Stages

研究不同分化阶段树突状细胞(dendritic cells,DCs)重要肌动蛋白微丝结合蛋白的表达变化。人外周血经密度梯度离心和免疫磁珠法分离获得CD14+单核细胞,用细胞因子将单核细胞(monocytes,MOs)诱导分化为未成熟DCs(immature DCs,im DCs)和成熟DCs(mature DCs,m DCs)。分别提取不同分化阶段DCs的总蛋白和总RNA,实时定量PCR和蛋白质芯片检测部分细胞骨架微丝(filament actin,F-actin)结合蛋白的在基因和蛋白水平的表达变化。单核细胞经im DCs向m DCs分化的过程中,一些F-actin的单体隔离蛋白、加帽蛋白、交联蛋白、解聚蛋白和成核蛋白在基因和蛋白水平发生了不同程度的上调或下调。一些重要的F-actin结合蛋白在DCs不同分化阶段具有不同的表达水平,DCs的结构和功能受到这些蛋白的协同作用和精密调控,是细胞形态发生显著变化的结构基础,这对于深入理解DCs的免疫调节功能来说具有重要意义。

To study the expression levels of important cytoskeleton-binding proteins in dendritic cells （DCs） at different differentiation stages, CD14^＋ monocytes （MOs） were separated from peripheral venous blood of healthy donors by Ficoll density gradient centrifugation and MACS, which was induced and differentiated by cell factors into immature DCs （imDCs） and mature DCs （mDCs）. The total protein and total RNA of DCs at different differentiation stages were extracted and the gene and protein expression levels of some important filament actin （F-actin）-binding proteins were measured by protein chip and Real-time PCR. In the process of MOs differentiated into mDCs via imDCs, the gene and protein expression levels of some monomer-isolated proteins, capping proteins, cross-linking proteins, actin-depolymerizing proteins and nucleating proteins of F-actin were up-regulated or down-regulated in various degrees. Some important F-actin binding proteins in DCs displayed various expression levels at different differentiation stages. The structures and functions of DCs were synergistically regulated by these proteins, which were the structural basis for remarkable cell morphological changes and was significant for the further understanding of the immune regulatory functions of DCs.