Characterization of the c-MYC-regulated transcriptome by SAGE: Identification and analysis of c-MYC target genes
Characterization of the c-MYC-regulated transcriptome by SAGE: Identification and analysis of c-MYC target genes
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DOI：
10.1073/pnas.082005599
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发表时间：
2002-04-30
影响因子：
11.1
通讯作者：
Hermeking, H
中科院分区：
文献类型：
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作者：
Menssen, A;Hermeking, H
为了鉴定致癌转录因子c-MYC的靶基因，我们在原代人脐静脉内皮细胞中腺病毒表达c-MYC后进行了基因表达序列分析（SAGE）：诱导了216种不同的SAGE标签，对应于独特的mrna，而表达c-MYC后抑制了260个标签（P < 0.05）。通过微阵列分析和实时定量PCR证实了53个基因的诱导作用：其中有MetAP2/p67基因，它编码翻译起始激活子，是抑制新生血管形成的有效靶点。此外，c-MYC诱导细胞周期调控基因CDC2-L1、Cyclin E结合蛋白1和Cyclin B1。DNA修复基因BRCA1、MSH2和APEX被c-MYC诱导，这表明c-MYC将DNA复制与保持基因组完整性的过程结合在一起。MNT，一种c-MYC功能的max结合拮抗剂，被上调，暗示一个负反馈回路。通过染色质免疫沉淀检测CDK4、Prohibitin、MNT、Cyclin B1和Cyclin E结合蛋白1在体内被c-MYC占用的启动子，表明这些基因是c-MYC的直接靶点。这里鉴定的c-MYC调控基因/标签将有助于确定一组真正的c-MYC靶点，并可能在抑制癌细胞增殖、肿瘤血管化和再狭窄方面具有潜在的治疗价值。
To identify target genes of the oncogenic transcription factor c-MYC, serial analysis of gene expression (SAGE) was performed after adenoviral expression of c-MYC in primary human umbilical vein endothelial cells: 216 different SAGE tags, corresponding to unique mRNAs, were induced, whereas 260 tags were repressed after c-MYC expression (P < 0.05). The induction of 53 genes was confirmed by using microarray analysis and quantitative real-time PCR: among these genes was MetAP2/p67, which encodes an activator of translational initiation and represents a validated target for inhibition of neovascularization. Furthermore, c-MYC induced the cell cycle regulatory genes CDC2-L1, Cyclin E binding protein 1, and Cyclin B1. The DNA repair genes BRCA1, MSH2, and APEX were induced by c-MYC, suggesting that c-MYC couples DNA replication to processes preserving the integrity of the genome. MNT, a MAX-binding antagonist of c-MYC function, was upregulated, implying a negative feedback loop. In vivo promoter occupancy by c-MYC was detected by chromatin immunoprecipitation for CDK4, Prohibitin, MNT, Cyclin B1, and Cyclin E binding protein 1, showing that these genes are direct c-MYC targets. The c-MYC-regulated genes/tags identified here will help to define the set of bona fide c-MYC targets and may have potential therapeutic value for inhibition of cancer cell proliferation, tumor-vascularization, and restenosis.