Genomic and proteomic profiling of oxidative stress response in human diploid fibroblasts.
Genomic and proteomic profiling of oxidative stress response in human diploid fibroblasts.
复制标题
人二倍体成纤维细胞中氧化应激反应的基因组和蛋白质组学分析。
DOI：
10.1007/s10522-008-9157-3
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发表时间：
2009-04
期刊：
影响因子：
4.5
通讯作者：
Chen QM
中科院分区：
文献类型：
--
作者：
Xie L;Pandey R;Xu B;Tsaprailis G;Chen QM
大量证据表明，培养物中正常人二倍体成纤维细胞（HDF）的衰老与体内衰老过程相关。我们使用正常的人类皮肤二倍体成纤维细胞，检查了用温和剂量的 H2O2 处理后基因和蛋白质的变化，H2O2 会导致过早衰老。多维蛋白质鉴定技术 (MudPIT) 结合 HDF 全细胞裂解物的质谱分析，检测到对照组中的 65 种蛋白质、H2O2 处理的细胞中的 48 种蛋白质以及两组中常见的 109 种蛋白质。相比之下，cDNA 微阵列分析显示 H2O2 处理后 173 个基因上调，179 个基因下调。 MudPIT 和 cDNA 微阵列分析均表明 H2O2 处理导致硫氧还蛋白还原酶 1 水平升高。半定量 RT-PCR 和 Western-blot 能够验证这一发现。在检测到的大量基因或蛋白质中，只有一小部分显示微阵列与蛋白质组学结果之间的重叠。低重叠表明在研究氧化应激改变的基因表达谱时考虑蛋白质而不是转录本的重要性。
A number of lines of evidence suggest that senescence of normal human diploid fibroblasts (HDFs) in culture is relevant to the process of aging in vivo. Using normal human skin diploid fibroblasts, we examine the changes in genes and proteins following treatment with a mild dose of H2O2, which induces premature senescence. Multidimensional Protein Identification Technology (MudPIT) in combination with mass spectrometry analyses of whole cell lysates from HDFs detected 65 proteins in control group, 48 proteins in H2O2-treated cells and 109 proteins common in both groups. In contrast, cDNA microarray analyses show 173 genes up-regulated and 179 genes down-regulated upon H2O2 treatment. Both MudPIT and cDNA microarray analyses indicate that H2O2 treatment caused elevated levels of thioredoxin reductase 1. Semi-quantitative RT-PCR and Western-blot were able to verify the finding. Out of a large number of genes or proteins detected, only a small fraction shows the overlap between the outcomes of microarray versus proteomics. The low overlap suggests the importance of considering proteins instead of transcripts when investigating the gene expression profile altered by oxidative stress.