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利用cDNA微阵列、蛋白质组学和基因本体分析对子宫平滑肌瘤进行靶向细胞过程分析的方法

Targeted cellular process profiling approach for uterine leiomyoma using cDNA microarray, proteomics and gene ontology analysis.

作者信息

Ahn Woong Shick, Kim Ko-Woon, Bae Su Mi, Yoon Joo Hee, Lee Joon Mo, Namkoong Sung Eun, Kim Jin Hong, Kim Chong Kook, Lee Young Joo, Kim Yong-Wan

机构信息

Department of Obstetrics and Gynecology, College of Medicine, The Catholic University of Korea, Seoul, South Korea.

出版信息

Int J Exp Pathol. 2003 Dec;84(6):267-79. doi: 10.1111/j.0959-9673.2003.00362.x.

Abstract

This study utilized both cDNA microarray and two-dimensional protein gel electrophoresis technology to investigate the multiple interactions of genes and proteins involved in uterine leiomyoma pathophysiology. Also, the gene ontology analysis was used to systematically characterize the global expression profiles at cellular process levels. We profiled differentially expressed transcriptome and proteome in six-paired leiomyoma and normal myometrium. Screening up to 17 000 genes identified 21 upregulated and 50 downregulated genes. The gene-expression profiles were classified into mutually dependent 420 functional sets, resulting in 611 cellular processes according to the gene ontology. Also, protein analysis using two-dimensional gel electrophoresis identified 33 proteins (17 upregulated and 16 downregulated) of more than 500 total spots, which was classified into 302 cellular processes. Of these functional profilings, downregulations of transcriptomes and proteoms were shown in cell adhesion, cell motility, organogenesis, enzyme regulator, structural molecule activity and response to external stimulus functional activities that are supposed to play important roles in pathophysiology. In contrast, the upregulation was only shown in nucleic acid-binding activity. Taken together, potentially significant pathogenetic cellular processes were identified and showed that the downregulated functional profiling has a significant impact on the discovery of pathogenic pathway in leiomyoma. Also, the gene ontology analysis can overcome the complexity of expression profiles of cDNA microarray and two-dimensional protein analysis via its cellular process-level approach. Therefore, a valuable prognostic candidate gene with relevance to disease-specific pathogenesis can be found at cellular process levels.

摘要

本研究利用cDNA微阵列和二维蛋白质凝胶电泳技术,研究子宫平滑肌瘤病理生理学中涉及的基因和蛋白质的多重相互作用。此外,基因本体分析用于在细胞过程水平上系统地表征全局表达谱。我们分析了六对平滑肌瘤和正常子宫肌层中差异表达的转录组和蛋白质组。筛选多达17000个基因,鉴定出21个上调基因和50个下调基因。基因表达谱被分类为相互依赖的420个功能集,根据基因本体,产生了611个细胞过程。此外,使用二维凝胶电泳的蛋白质分析在总共500多个斑点中鉴定出33种蛋白质(17种上调和16种下调),这些蛋白质被分类为302个细胞过程。在这些功能分析中,转录组和蛋白质组的下调在细胞粘附、细胞运动、器官发生、酶调节、结构分子活性和对外部刺激的反应等功能活动中表现出来,这些功能活动在病理生理学中应该发挥重要作用。相比之下,上调仅在核酸结合活性中表现出来。综上所述,确定了潜在的重要致病细胞过程,表明下调的功能分析对平滑肌瘤致病途径的发现有重大影响。此外,基因本体分析可以通过其细胞过程水平的方法克服cDNA微阵列和二维蛋白质分析表达谱的复杂性。因此,在细胞过程水平上可以找到与疾病特异性发病机制相关的有价值的预后候选基因。

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