Comparative proteomic phenotyping of cell lines and primary cells to assess preservation of cell type-specific functions.
Comparative proteomic phenotyping of cell lines and primary cells to assess preservation of cell type-specific functions.
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DOI：
10.1074/mcp.m800258-mcp200
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发表时间：
2009-03
期刊：
影响因子：
--
通讯作者：
Mann M
中科院分区：
文献类型：
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作者：
Pan C;Kumar C;Bohl S;Klingmueller U;Mann M
生物学实验最常使用永生化细胞系进行，因为它们容易获得并且可以无限制地扩增。然而，细胞系在重要方面可能与体内情况不同。在这里，我们介绍了一个简单的方法来比较细胞系，其同源的原代细胞，并得出一个比较功能表型。我们使用SILAC（细胞培养物中氨基酸的稳定同位素标记）对肝癌细胞系Hepa 1 -6与原代肝细胞进行基于质谱的定量比较。4，063种蛋白质的定量蛋白质组具有不对称分布，其中许多蛋白质在细胞系中下调。定量蛋白质组学表型的生物信息学分析显示，Hepa 1 -6细胞缺乏线粒体，反映了代谢途径的重排，大幅上调细胞周期相关功能，并在很大程度上关闭了肝脏特有的药物代谢酶。这种变化的定量知识为使细胞系适应更接近生理条件提供了重要基础。
Biological experiments are most often performed with immortalized cell lines because they are readily available and can be expanded without limitation. However, cell lines may differ from the in vivo situation in important aspects. Here we introduce a straightforward methodology to compare cell lines to their cognate primary cells and to derive a comparative functional phenotype. We used SILAC (stable isotope labeling by amino acids in cell culture) for quantitative, mass spectrometry-based comparison of the hepatoma cell line Hepa1–6 with primary hepatocytes. The resulting quantitative proteome of 4,063 proteins had an asymmetric distribution, with many proteins down-regulated in the cell line. Bioinformatic analysis of the quantitative proteomics phenotypes revealed that Hepa1–6 cells were deficient in mitochondria, reflecting re-arrangement of metabolic pathways, drastically up-regulate cell cycle-associated functions and largely shut down drug metabolizing enzymes characteristic for the liver. This quantitative knowledge of changes provides an important basis to adapt cell lines to more closely resemble physiological conditions.