Global analysis of HuR-Regulated gene expression in colon cancer systems of reducing complexity
Global analysis of HuR-Regulated gene expression in colon cancer systems of reducing complexity
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DOI：
10.3727/000000004783992215
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发表时间：
2004-01-01
期刊：
影响因子：
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通讯作者：
Gorospe, M
中科院分区：
文献类型：
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作者：
De Silanes, IL;Fan, JS;Gorospe, M
HuR是一种与靶mRNA结合并能增强其稳定性和翻译的蛋白质，越来越多地被认为是细胞分裂和肿瘤发生过程中基因表达的关键调节因子。我们试图通过系统的、基于cDNA阵列的基因表达评估来确定结肠癌细胞中HuR调控的mRNA的集合，这些评估在三个不同复杂性的系统中进行。首先，不同HuR丰度的肿瘤之间的基因表达谱的比较揭示了高度不同的基因表达模式，并且在以前报道的HuR靶mRNA中几乎没有变化。在第二个降低复杂性的系统中评估基因表达模式，培养的结肠癌细胞表达不同的HuR水平，呈现出更保守的HuR调节的mRNA集。然而，直接HuR靶mRNA的确定性鉴定需要复杂性更低的第三个系统，其中从结肠癌细胞免疫沉淀的HuR-RNA复合物进行cDNA阵列杂交以阐明内源性HuR结合的mRNA。在每个系统中确定的转录本集的比较揭示了一个惊人的有限的重叠HuR调节的mRNA。从HuR调节基因的系统分析中获得的数据突出了蛋白质-RNA相互作用的低复杂性，生物化学表征的价值。然而，更重要的是，这些数据强调了包括低复杂性（蛋白质-核酸）和高复杂性（细胞、肿瘤）系统的综合方法的广泛用途，以全面阐明生物过程的基因调控事件。
HuR, a protein that binds to target mRNAs and can enhance their stability and translation, is increasingly recognized as a pivotal regulator of gene expression during cell division and tumorigenesis. We sought to identify collections of HuR-regulated mRNAs in colon cancer cells by systematic, cDNA array-based assessment of gene expression in three systems of varying complexity. First, comparison of gene expression profiles among tumors with different HuR abundance revealed highly divergent gene expression patterns, and virtually no changes in previously reported HuR target mRNAs. Assessment of gene expression patterns in a second system of reduced complexity, cultured colon cancer cells expressing different HuR levels, rendered more conserved sets of HuR-regulated mRNAs. However, the definitive identification of direct HuR target mRNAs required a third system of still lower complexity, wherein HuR-RNA complexes immunoprecipitated from colon cancer cells were subject to cDNA array hybridization to elucidate the endogenous HuR-bound mRNAs. Comparison of the transcript sets identified in each system revealed a strikingly limited overlap in HuR-regulated mRNAs. The data derived from this systematic analysis of HuR-regulated genes highlight the value of low-complexity, biochemical characterization of protein-RNA interactions. More importantly, however, the data underscore the broad usefulness of integrated approaches comprising systems of low complexity (protein-nucleic acid) and high complexity (cells, tumors) to comprehensively elucidate the gene regulatory events that underlie biological processes.