Medical Proteomics Unit, Office for Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok, Thailand.
J Proteomics. 2010 Jan 3;73(3):602-18. doi: 10.1016/j.jprot.2009.08.001. Epub 2009 Aug 12.
Monocyte and macrophage are mainly involved in immune response and inflammatory processes. Monocytes circulate in the bloodstream and migrate to various tissues where they can differentiate to macrophages. However, the molecular basis of biological processes involved in this cellular differentiation remains ambiguous. This study was to investigate alterations in cellular and secreted proteins after this differentiation phase. Macrophage was differentiated from U937 human monocytic cell line by treatment with 100 ng/ml phorbol myristate acetate (PMA) for 48 h. Cellular and secreted proteins extracted from PMA-treated cells (macrophages) were compared with those of untreated cells (monocytes) using 2-DE (n=5 gels/condition; stained with Deep Purple fluorescence dye). Quantitative intensity analysis revealed 81 and 67 protein spots whose levels were significantly altered in cellular proteome and secretome. These proteins were subsequently identified by Q-TOF MS and/or MS/MS analyses. The altered levels of cellular elongation factor-2 (EF-2) and secreted alpha-tubulin were confirmed by Western blot analysis. Global protein network analysis demonstrated that these altered proteins were involved in cell death, lipid metabolism, cell morphology, cellular movement, and protein folding. Our data may provide some insights into molecular mechanisms of biological processes upon differentiation from monocytes to macrophages.
单核细胞和巨噬细胞主要参与免疫反应和炎症过程。单核细胞在血液中循环，并迁移到各种组织中，在那里它们可以分化为巨噬细胞。然而，参与这种细胞分化的生物学过程的分子基础仍然不清楚。本研究旨在研究分化阶段后细胞和分泌蛋白的变化。用 100ng/ml 佛波酯（PMA）处理 U937 人单核细胞系 48 小时，将巨噬细胞从单核细胞中分化而来。使用 2-DE（n=5 个凝胶/条件；用 Deep Purple 荧光染料染色）比较 PMA 处理的细胞（巨噬细胞）和未经处理的细胞（单核细胞）中的细胞和分泌蛋白。定量强度分析显示细胞伸长因子-2（EF-2）和分泌的α-微管蛋白的水平发生显著变化，细胞蛋白质组和分泌蛋白质组中分别有 81 和 67 个蛋白斑点。随后通过 Q-TOF MS 和/或 MS/MS 分析鉴定这些蛋白质。Western blot 分析证实了细胞内延伸因子-2（EF-2）和分泌的α-微管蛋白水平的变化。蛋白质整体网络分析表明，这些变化的蛋白质参与细胞死亡、脂质代谢、细胞形态、细胞运动和蛋白质折叠。我们的数据可能为单核细胞向巨噬细胞分化过程中的分子机制提供一些见解。
