Hepatocyte nuclear factor 1 coordinates multiple processes in a model of intestinal epithelial cell function.
Hepatocyte nuclear factor 1 coordinates multiple processes in a model of intestinal epithelial cell function.
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肝细胞核因子 1 协调肠上皮细胞功能模型中的多个过程。
DOI：
10.1016/j.bbagrm.2016.02.005
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Harris,Ann
中科院分区：
文献类型：
--
作者：
Yang,Rui;Kerschner,JennyL;Harris,Ann
肝细胞核因子1转录因子(HNF1α/β)突变与糖尿病有关。这些因子在肝脏、胰腺和肾脏中得到了很好的研究，在那里它们指导着组织特异性的基因调控。然而，它们在包括肠道在内的许多其他组织的生物学中也扮演着重要的角色。我们研究了HNF1在肠上皮细胞系(Caco 2)中调控的转录网络。我们使用染色质免疫沉淀和直接测序(CHIP-SEQ)来鉴定全基因组范围内的HNF1结合位点。用传统的芯片方法验证HNF1的直接靶点，并通过siRNA介导的HNF1缺失和RT-qPCR确认。对HNF1靶标的基因本体论过程丰富分析确定了在肠道上皮细胞功能中起作用的多个过程，包括细胞膜的属性、细胞对激素的反应以及生物合成过程的调节。大约50%的HNF1结合位点也被肠道转录网络的其他成员占据，包括肝细胞核因子4A(HNF4a)、尾型同源盒2(CDX2)和叉头盒A2(FOXA2)。Caco 2细胞中HNF1的缺失增加了FOXA2的丰度，降低了CDX2的水平，说明了该网络的协调活动。这些数据表明，HNF1在调节肠道上皮细胞功能方面发挥着重要作用，无论是直接还是通过与其他肠道转录因子的相互作用。
Mutations in hepatocyte nuclear factor 1 transcription factors (HNF1α/β) are associated with diabetes. These factors are well studied in the liver, pancreas and kidney, where they direct tissue-specific gene regulation. However, they also have an important role in the biology of many other tissues, including the intestine. We investigated the transcriptional network governed by HNF1 in an intestinal epithelial cell line (Caco2). We used chromatin immunoprecipitation followed by direct sequencing (ChIP-seq) to identify HNF1 binding sites genome-wide. Direct targets of HNF1 were validated using conventional ChIP assays and confirmed by siRNA-mediated depletion of HNF1, followed by RT-qPCR. Gene ontology process enrichment analysis of the HNF1 targets identified multiple processes with a role in intestinal epithelial cell function, including properties of the cell membrane, cellular response to hormones, and regulation of biosynthetic processes. Approximately 50% of HNF1 binding sites were also occupied by other members of the intestinal transcriptional network, including hepatocyte nuclear factor 4A (HNF4A), caudal type homeobox 2 (CDX2), and forkhead box A2 (FOXA2). Depletion of HNF1 in Caco2 cells increases FOXA2 abundance and decreases levels of CDX2, illustrating the coordinated activities of the network. These data suggest that HNF1 plays an important role in regulating intestinal epithelial cell function, both directly and through interactions with other intestinal transcription factors.