DNA microarrays identification of primary and secondary target genes regulated by p53
DNA microarrays identification of primary and secondary target genes regulated by p53
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DOI：
10.1038/sj.onc.1204319
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发表时间：
2001-04-26
期刊：
影响因子：
8
通讯作者：
Givol, D
中科院分区：
文献类型：
--
作者：
Kannan, K;Amariglio, N;Givol, D
利用寡核苷酸微阵列分析了肿瘤抑制基因p53调控的转录程序。利用表达温度敏感性鼠p53的人肺癌细胞系来定量在将温度转移到32 ℃后的不同时间点的各种基因的mRNA水平。放线菌酮（CHX）对蛋白质合成的抑制作用被用来区分p53调控的主要和次要靶基因。在不存在CHX的情况下，分别有259和125个基因上调或下调;这些基因中只有38和24个基因在存在CHX的情况下也被p53上调和下调，并且被认为是该细胞系中的主要靶标。这些数据的聚类分析，使用超顺磁性聚类（SPC）算法表明，主要基因可以区分为一个单一的集群之间的一个大池的p53调控基因。该程序鉴定了与主要靶标共簇的另外的基因，并且也可以被分类为这样的基因。（例如p21、TGF-β、细胞周期蛋白E）和细胞凋亡（例如，Fas、巴克、IAP）相关基因，p53的主要靶标包括涉及细胞功能的许多方面的基因，包括细胞粘附（例如，胸腺素、Smoothelin）、信号传导（例如H-Ras，二酰基甘油激酶），转录（例如，ATF 3、LISCH 7）、神经元生长（例如Ninjurin、NSCL2）和DNA修复（例如BTG2、DDB2），结果表明，p53激活协调的相反信号，并通过多种转录变化网络发挥其作用，改变细胞表型以响应压力。
The transcriptional program regulated by the tumor suppressor p53 was analysed using oligonucleotide microarrays. A human lung cancer cell line that expresses the temperature sensitive murine p53 was utilized to quantitate mRNA levels of various genes at different time points after shifting the temperature to 32 degreesC. Inhibition of protein synthesis by cycloheximide (CHX) was used to distinguish between primary and secondary target genes regulated by p53. In the absence of CHX, 259 and 125 genes were up or down-regulated respectively; only 38 and 24 of these genes were up and down-regulated by p53 also in the presence of CHX and are considered primary targets in this cell line. Cluster analysis of these data using the super paramagnetic clustering (SPC) algorithm demonstrate that the primary genes can be distinguished as a single cluster among a large pool of p53 regulated genes. This procedure identified additional genes that co-cluster with the primary targets and can also be classified as such genes, In addition to cell cycle (e.g. p21, TGF-beta, Cyclin E) and apoptosis (e.g, Fas, Bak, IAP) related genes, the primary targets of p53 include genes involved in many aspects of cell function, including cell adhesion (e.g, Thymosin, Smoothelin), signaling (e.g. H-Ras, Diacylglycerol kinase), transcription (e.g. ATF3, LISCH7), neuronal growth (e.g. Ninjurin, NSCL2) and DNA repair (e.g. BTG2, DDB2), The results suggest that p53 activates concerted opposing signals and exerts its effect through a diverse network of transcriptional changes that collectively alter the cell phenotype in response to stress.