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芯片免疫共沉淀法分析胃癌细胞中 HMGA2 转录因子结合位点的全基因组。

Genome-wide analysis of HMGA2 transcription factor binding sites by ChIP on chip in gastric carcinoma cells.

机构信息

Department of Gastrointestinal Surgery, First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, People's Republic of China.

出版信息

Mol Cell Biochem. 2012 May;364(1-2):243-51. doi: 10.1007/s11010-012-1224-z. Epub 2012 Jan 14.

Abstract

High mobility group protein A2 (HMGA2) is an architectural transcription factor that plays an important role in development and progression of malignant neoplasias. Recently, some studies reported that HMGA2 is also implicated in epithelial-mesenchymal transitions (EMT) and cancer stem cells. But the underlying mechanisms of these conditions are poorly understood. Therefore, we established an EMT model of gastric carcinoma cells by overexpressing HMGA2 in vitro, then global mapping of HMGA2 potential transcription factor binding sites was identified by promoter microarray in these cells, and the date obtained from the microarrays were validated via chromatin immunoprecipitation-PCR (ChIP-PCR) and real-time PCR. HMGA2 potential target genes were classified in KEGG database and Gene Ontology (GO) analyses. To our knowledge, this is the first report on the genome-wide analysis of HMGA2 downstream direct targets, and these findings will be valuable in understanding the roles of HMGA2 in EMT.

摘要

高迁移率族蛋白 A2(HMGA2)是一种结构转录因子,在恶性肿瘤的发生和发展中起着重要作用。最近的一些研究表明,HMGA2 也与上皮-间充质转化(EMT)和癌症干细胞有关。但是,这些情况的潜在机制尚不清楚。因此,我们通过体外过表达 HMGA2 建立了胃癌细胞的 EMT 模型,然后通过这些细胞的启动子微阵列鉴定了 HMGA2 潜在转录因子结合位点的全局图谱,并通过染色质免疫沉淀-PCR(ChIP-PCR)和实时 PCR 验证了从微阵列获得的数据。HMGA2 潜在的靶基因被分类在 KEGG 数据库和基因本体论(GO)分析中。据我们所知,这是关于 HMGA2 下游直接靶基因的全基因组分析的首次报道,这些发现将有助于理解 HMGA2 在 EMT 中的作用。

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