Development of cDNA microarray for expression profiling of estrogen-responsive genes
Development of cDNA microarray for expression profiling of estrogen-responsive genes
复制标题
DOI：
10.1677/jme.0.0290175
复制
发表时间：
2002-10-01
影响因子：
3.5
通讯作者：
Hayashi, S
中科院分区：
文献类型：
--
作者：
Inoue, A;Yoshida, N;Hayashi, S
雌激素在许多生理事件中起着重要作用，包括致癌和人类乳腺癌的发展。但是，迄今尚未阐明癌症中雌激素信号传导的分子机制，并且需要对乳腺癌的准确治疗预测。我们首先在雌激素受体阳性人MCF-7乳腺癌细胞中对大约9000个基因进行了雌激素反应性表达分析。根据结果，选择雌激素反应基因以生产定制的cDNA微阵列。使用由缩小基因子群组成的微阵列，我们首先分析了MCF-7细胞中雌激素反应性基因表达谱的时间过程，从而导致雌激素在早期和晚期 - 反应性和晚期 - 反应性和晚期中的上调。响应基因。通过北印迹分析证实了几种基因的表达模式。我们还分析了雌激素拮抗剂ICI 182,780和4-羟基莫昔芬（OHT）对MCF-7细胞中雌激素反应基因表达谱的影响。虽然雌激素对大多数基因的调节被V 182,780完全消除了，但即使在OHT存在的情况下，某些基因也受到雌激素的部分调节。此外，通过层次聚类（包括MCF-7细胞中的特征）获得了源自乳腺，卵巢，胃和其他组织的十二个癌细胞系的雌激素反应基因表达谱。除MCF-7细胞以外的其他细胞系中，雌激素还表现出几种基因的上调或下调。讨论了这些分析中关于癌症性质的雌激素反应基因的重要性。
Estrogen plays an important role in many physiological events including carcinogenesis and the development of human breast cancer. However, the molecular mechanisms of estrogen signaling in cancers have not been clarified hitherto and accurate therapeutic prediction of breast cancer is earnestly desired. We first carried out estrogen-responsive expression profiling of approximately 9000 genes in estrogen receptor-positive human MCF-7 breast cancer cells. Based on the results, estrogen-responsive genes were selected for production of a custom-made cDNA microarray. Using a microarray consisting of the narrowed-down gene subset, we first analyzed the time course of the estrogen-responsive gene expression profiles in MCF-7 cells, resulting in subdivision of the genes up-regulated by estrogen into early-responsive and late-responsive genes. The expression patterns of several genes were confirmed by Northern blot analysis. We also analyzed the effects of the estrogen antagonists ICI 182,780 and 4-hydroxytamoxifen (OHT) on the estrogen-responsive gene expression profiles in MCF-7 cells. While the regulation of most of the genes by estrogen was completely abolished by V 182,780, some genes were partially regulated by estrogen even in the presence of OHT. Furthermore, the estrogen-responsive gene expression profiles of twelve cancer cell lines derived from the breast, ovary, stomach and other tissues were obtained and analyzed by hierarchical clustering including the profiles in MCF-7 cells. Several genes also showed up-regulation or down-regulation by estrogen in cell lines other than MCF-7 cells. The significance of the estrogen-responsive genes identified in these analyses concerning the nature of cancer is discussed.