A combination of proteomics, principal component analysis and transcriptomics is a powerful tool for the identification of biomarkers for macrophage maturation in the U937 cell line
A combination of proteomics, principal component analysis and transcriptomics is a powerful tool for the identification of biomarkers for macrophage maturation in the U937 cell line
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蛋白质组学、主成分分析和转录组学的结合是鉴定 U937 细胞系中巨噬细胞成熟生物标志物的强大工具
作者：
K. Verhoeckx;S. Bijlsma;E. M. de Groene;R. Witkamp;J. van der Greef;R. Rodenburg
单核细胞样人组织细胞性淋巴瘤细胞系U937可以被佛波醇12-肉豆蔻酸酯13-乙酸酯（PMA）诱导分化为巨噬细胞样表型。我们使用二维凝胶电泳（2-DE）、寡核苷酸微阵列和主成分分析（PCA）来表征U937细胞系作为单核细胞分化为巨噬细胞的模型系统。共发现226个差异表达蛋白，其中41个通过PCA选择用于使用基质辅助激光解吸/电离串联质谱鉴定。基于PCA结果，选择三种标记蛋白，用于使用Western印迹和定量真实的时间PCR确认差异表达。选择的标志物蛋白为：γ干扰素诱导型溶酶体巯基还原酶、组织蛋白酶D和脂肪细胞-脂肪酸结合蛋白。所有三种都被证明是U937细胞以及外周血来源的巨噬细胞的巨噬细胞成熟的良好分化标志物。转录组学数据揭示了U937巨噬细胞中大量额外的推定分化标志物，其中许多已知在外周血来源的巨噬细胞中表达。这些包括骨脊椎蛋白、基质金属蛋白酶9和HC-gp 39。我们的研究结果表明，U937巨噬细胞的特征类似于炎性（渗出液）巨噬细胞的特征，例如5'核苷酸酶的下调和亮氨酸氨肽酶mRNA的上调。总之，使用转录组学、2-DE和PCA的强大组合，我们的结果表明，通过PMA处理分化的U937细胞是从血液产生单核细胞衍生的巨噬细胞的极好模型系统。
The monocyte‐like human histiocytic lymphoma cell line U937 can be induced by phorbol 12‐myristate 13‐acetate (PMA) to undergo differentiation into a macrophage‐like phenotype. We have used two‐dimensional gel electrophoresis (2‐DE), oligonucleotide microarrays and principal component analysis (PCA) to characterize the U937 cell line as a model system for the differentiation of monocytes into macrophages. A total of 226 differentially expressed proteins were found, of which 41 were selected by PCA for identification using matrix‐assisted laser desorption/ionization tandem mass spectrometry. Based on the PCA results, three marker proteins were selected for confirmation of differential expression using Western blot and quantitative real time‐PCR. The selected marker proteins were: gamma interferon inducible lysosomal thiol reductase, cathepsin D and adipocyte‐fatty acid binding protein. All three proved to be good differentiation markers for macrophage maturation of U937 cells as well as peripheral blood‐derived macrophages. The transcriptomics data revealed a large number of additional putative differentiation markers in U937 macrophages, many of which are known to be expressed in peripheral blood‐derived macrophages. These include osteospontin, matrix metalloproteinase 9, and HC‐gp39. Our results show that the characteristics of U937 macrophages resemble those of inflammatory (exudate) macrophages, exemplified by the down‐regulation of 5' nucleotidase and the up‐regulation of leucine aminopeptidase mRNAs. In conclusion, using the powerful combination of transcriptomics, 2‐DE and PCA, our results show that U937 cells differentiated by PMA treatment are an excellent model system for monocyte derived macrophage generation from blood.