Catholic Research Institutes of Medical Science, College of Medicine, The Catholic University of Korea, Seoul, Korea.
Cancer Res Treat. 2004 Feb;36(1):31-42. doi: 10.4143/crt.2004.36.1.31. Epub 2004 Feb 29.
This study utilized both cDNA microarray and 2D protein gel electrophoresis technology to investigate the multiple interactions of the genes and proteins involved in the pathophysiology of uterine leiomyomas. Also, Gene Ontology (GO) analysis was used to systematically characterize the global expression profiles, which were found to correlate with the leiomyosarcomas.
The uterine leiomyoma biopsies were obtained from patients in the Department of Obstetrics and Gynecology, The Catholic University of Korea. Differentially expressed transcriptome and proteome, in 6 paired leiomyoma and normal myometrium, were profiled. The total RNAs from the leiomyoma and normal myometrium were labeled with Cy5 and Cy3. All specimens were punch-biopsy-obtained, and frozen in liquid nitrogen.
Screening of up to 17,000 genes identified 71 that were either up-regulated or down-regulated (21 and 50, respectively). The gene expression profiles were classified into 420 mutually dependent functional sets, resulting in 611 cellular processes, according to the gene ontology. Also, the protein analysis, using 2D gel electrophoresis, identified 33 proteins (17 up-regulated and 16 down-regulated) with more than 500 total spots, which were classified into 302 cellular processes. O f these functional profilings, transcriptomes and proteoms down-regulations were shown in the cell adhesion, cell motility, organogenesis, enzyme regulator, structural molecule activity and responses to external stimulus functional activities, which are supposed to play important roles in the pathophysiology. In contrast, up-regulation was only shown in the nucleic acid binding activity. The CDKN2A, ADH1A, DCX, IGF2, CRABP2 and KIF5C were found to increase the reliability of this study, and correlate with the leiomyosarcomas.
Potentially significant pathogenetic cellular processes showed that down-regulated functional profiling has an important impact on the discovery of the pathogenic pathways in leiomyomas and leiomyosarcomas. GO analysis can also overcome the complexity of the expression profiles of cDNA microarrays and 2D protein analyses, via a cellular process level approach. Thereby, a valuable prognostic candidate gene, with real relevance to disease-specific pathogenesis, can be found at cellular process levels.
本研究利用 cDNA 微阵列和 2D 蛋白质凝胶电泳技术来研究涉及子宫肌瘤病理生理学的基因和蛋白质的多种相互作用。此外，还使用基因本体论 (GO) 分析对全局表达谱进行了系统表征，发现这些表达谱与平滑肌瘤肉瘤相关。
从韩国天主教大学妇产科获得子宫肌瘤活检。对 6 对子宫肌瘤和正常子宫肌层的差异表达转录组和蛋白质组进行了分析。将来自子宫肌瘤和正常子宫肌层的总 RNA 用 Cy5 和 Cy3 标记。所有标本均通过打孔活检获得，并在液氮中冷冻。
筛选了多达 17000 个基因，发现了 71 个上调或下调的基因（分别为 21 个和 50 个）。根据基因本体论，将基因表达谱分类为 420 个相互依赖的功能集，导致 611 个细胞过程。此外，使用 2D 凝胶电泳进行的蛋白质分析鉴定了 33 种蛋白质（17 种上调和 16 种下调），总斑点超过 500 个，这些蛋白质分为 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显示在细胞黏附、细胞运动、器官发生、酶调节剂、结构分子活性和对外界刺激的反应等功能活动中，这些功能活动可能在病理生理学中发挥重要作用。相比之下，上调仅显示在核酸结合活性中。CDKN2A、ADH1A、DCX、IGF2、CRABP2 和 KIF5C 的表达增加了本研究的可靠性，并与平滑肌瘤肉瘤相关。
潜在的重要致病细胞过程表明，下调的功能分析对发现子宫肌瘤和平滑肌瘤肉瘤的致病途径具有重要影响。GO 分析还可以通过细胞过程水平的方法克服 cDNA 微阵列和 2D 蛋白质分析表达谱的复杂性。因此，可以在细胞过程水平找到与疾病特异性发病机制真正相关的有价值的预后候选基因。
