The Biotechnology Centre of Oslo and Centre for Molecular Medicine, Nordic EMBL Partnership, University of Oslo, Oslo, Norway.
Cell Signal. 2011 Jan;23(1):14-8. doi: 10.1016/j.cellsig.2010.07.009. Epub 2010 Jul 16.
Analysis of signalling events by classical biochemical approaches is limited as the outcome is an averaged readout for protein activation of a single protein within a cell population. This is a clear restriction when addressing signalling events in mixed populations or subpopulations of cells. By combining flow cytometry with a panel of phosphospecific antibodies against several signal molecules simultaneously in a multi-parameter phospho flow cytometry analysis it is possible to obtain a higher level of understanding of the signal transduction dynamics at a single cell level. In addition, analysis of mixed cell populations makes it possible to study cells ex vivo in a state more closely resembling the in vivo situation. The multimeric analysis yields information on combinations of signals turned on and off in specific settings such as disease (signal nodes) that can be used for biomarker analysis and for development of drug screening strategies. Prostaglandin E(2) (PGE(2)) is known to signal through four G-protein coupled transmembrane receptors, EP1-4, activating a multitude of potential signalling pathways. The analysis of the PGE(2) signalling network elicited by activation of the four EP receptors in lymphoid cells revealing several signalling nodes is reviewed as an example.
通过经典的生化方法分析信号事件受到限制，因为其结果是对细胞群体中单种蛋白的蛋白激活的平均读出。当在混合群体或细胞亚群中处理信号事件时，这是一个明显的限制。通过将流式细胞术与针对几种信号分子的磷酸特异性抗体组合，同时进行多参数磷酸化流式细胞术分析，可以在单细胞水平上获得对信号转导动力学的更深入理解。此外，对混合细胞群的分析使得有可能在更接近体内情况的状态下研究离体细胞。多聚体分析提供了关于在特定环境（信号节点）中打开和关闭的信号组合的信息，可用于生物标志物分析和药物筛选策略的开发。前列腺素 E2 (PGE2) 已知通过四个 G 蛋白偶联跨膜受体 EP1-4 发出信号，激活多种潜在的信号通路。本文以激活淋巴细胞中的四个 EP 受体引发的 PGE2 信号网络分析为例进行综述，揭示了几个信号节点。
