Microarray analysis of E-box binding-related gene expression in young and replicatively senescent human fibroblasts.
Microarray analysis of E-box binding-related gene expression in young and replicatively senescent human fibroblasts.
复制标题
年轻和复制性衰老人类成纤维细胞中 E-box 结合相关基因表达的微阵列分析。
DOI：
10.1006/abio.2001.5515
复制
发表时间：
2002
影响因子：
2.9
通讯作者：
Wang,Eugenia
中科院分区：
文献类型：
--
作者：
Semov,Alexandre;Marcotte,Richard;Semova,Natalie;Ye,Xiangyun;Wang,Eugenia
开发了 E-box (CACGTG) 设计微阵列来监测一组其表达具有特定调控模式的基因。评估了微阵列杂交的敏感性和特异性以及微阵列数据的变异性。该设计微阵列用于在三种不同生长状态的 WI-38 成纤维细胞培养物中生成 E-box 结合相关基因的表达谱：低传代复制、低传代接触抑制静止和复制衰老。微阵列基因筛选显示，与复制细胞相比，静止和衰老细胞的特征是 Pam（一种与 c-Myc 相关的蛋白质）下调，以及 Mad 家族基因 Max 二聚化蛋白上调。此外，静止和衰老可以通过 Irlb（c-Myc 转录因子）和 Miz-1（c-Myc 相互作用锌指蛋白 1）的表达增加来区分（仅在前一种状态下）。与年轻的复制状态和静止状态相比，衰老的特征是 Id4（DNA 结合 4 的抑制剂）和 Mitf（小眼球相关转录因子）的下调。通过逆转录聚合酶链式反应技术独立证实了微阵列杂交检测到的基因的差异表达。 E-box结合转录因子和c-Myc结合蛋白表达的改变证明了这些基因在建立接触抑制的静止或衰老表型中的重要性。
An E-box (CACGTG) designer microarray was developed to monitor a group of genes whose expressions share a particular regulatory mode. Sensitivity and specificity of microarray hybridization, as well as variability of microarray data, were evaluated. This designer microarray was used to generate expression profiles of E-box binding-related genes in WI-38 fibroblast cultures at three different growth states: low-passage replicating, low-passage contact-inhibited quiescent, and replicatively senescent. Microarray gene screening reveals that quiescent and senescent cells, in comparison with replicating ones, are characterized by downregulation of Pam, a protein associated with c-Myc, and upregulation of Mad family genes, Max dimerization proteins. Moreover, quiescence and senescence can be distinguished by increased expression of Irlb, c-Myc transcription factor, and Miz-1, c-Myc-interacting Zn finger protein 1, only in the former state. Senescence is characterized by downregulation of Id4, inhibitor of DNA binding 4, and Mitf, microphthalmia-associated transcription factor, in comparison with young replicating and quiescent states. Differential expression of genes detected by microarray hybridization was independently confirmed by reverse transcription polymerase chain reaction technique. Alterations in the expression of E-box-binding transcription factors and c-Myc-binding proteins demonstrate the importance of these genes in establishing the contact-inhibited quiescent or senescent phenotypes.