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Differential Expression of Extracellular Matrix Proteins in Senescent and Young Human Fibroblasts: a Comparative Proteomics and Microarray Study

机译:衰老和年轻的人类成纤维细胞中细胞外基质蛋白的差异表达:比较蛋白质组学和芯片研究。

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摘要

The extracellular matrix (ECM) provides an essential structural framework for cell attachment, proliferation, and differentiation, and undergoes progressive changes during senescence. To investigate changes in protein expression in the extracellular matrix between young and senescent fibroblasts, we compared proteomic data (LTQ-FT) with cDNA microarray results. The peptide counts from the proteomics analysis were used to evaluate the level of ECM protein expression by young cells and senescent cells, and ECM protein expression data were compared with the microarray data. After completing the comparative analysis, we grouped the genes into four categories. Class I included genes with increased expression levels in both analyses, while class IV contained genes with reduced expression in both analyses. Class II and Class III contained genes with an inconsistent expression pattern. Finally, we validated the comparative analysis results by examining the expression level of the specific gene from each category using Western blot analysis and semiquantitative RT-PCR. Our results demonstrate that comparative analysis can be used to identify differentially expressed genes.
机译:细胞外基质(ECM)为细胞附着,增殖和分化提供了必不可少的结构框架,并在衰老过程中进行性变化。为了研究年轻和衰老的成纤维细胞在细胞外基质中蛋白质表达的变化,我们将蛋白质组学数据(LTQ-FT)与cDNA微阵列结果进行了比较。蛋白质组学分析中的肽计数用于评估年轻细胞和衰老细胞的ECM蛋白表达水平,并将ECM蛋白表达数据与微阵列数据进行比较。完成比较分析后,我们将基因分为四类。 I类包括两种分析中表达水平升高的基因,而IV类包含两种分析中表达水平降低的基因。 II类和III类包含表达模式不一致的基因。最后,我们通过使用蛋白质印迹分析和半定量RT-PCR检查各类别中特定基因的表达水平,验证了比较分析结果。我们的结果表明,比较分析可用于鉴定差异表达的基因。

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