The CCAAT box binding transcription factor, nuclear factor-Y (NF-Y) regulates transcription of human aldo-keto reductase 1C1 (AKR1C1) gene.
The CCAAT box binding transcription factor, nuclear factor-Y (NF-Y) regulates transcription of human aldo-keto reductase 1C1 (AKR1C1) gene.
复制标题
CCAAT 盒结合转录因子核因子 Y (NF-Y) 调节人醛酮还原酶 1C1 (AKR1C1) 基因的转录。
DOI：
10.1016/j.gene.2010.03.006
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发表时间：
2010
期刊：
影响因子：
3.5
通讯作者：
Parekh,HemantK
中科院分区：
文献类型：
--
作者：
Pallai,Rajash;Simpkins,Henry;Chen,Jianli;Parekh,HemantK
二氢二醇脱氢酶是醛酮还原酶（AKR 1C）家族，参与类固醇激素和异生物质的代谢。在此，我们已经克隆并表征了人类AKR 1C 1基因的近端启动子区。AKR 1C 1基因的5′侧翼近端启动子区由TATA盒和反向CCAAT结合位点组成。对人类AKR 1C 1基因5′侧翼约3.0kb区域的缺失分析发现，-128和-88之间的区域是人类卵巢癌（2008和2008/C13*）、肺癌（H23和A549）和肝癌（HepG 2）细胞中AKR 1C 1基础转录所必需的最小近端启动子。定点突变研究表明，在所有研究的癌细胞中，NF-Y/CEBP的转录因子结合位点参与控制AKR 1C 1的基础转录。电泳迁移率变动（EMSA）和凝胶超移分析表明，转录因子NF-Y优先结合AKR 1C 1基因的-109ATTGG-105处的反向CCAAT盒。染色质免疫沉淀（ChIP）分析证实了在人卵巢癌、肺癌和肝癌细胞中NF-Y和人AKR 1C 1基因启动子之间的体内关联。NF-Ys的异位表达增加了AKR 1C 1基因的转录，而显性负性NF-YA的表达或抑制NF-YA则降低了AKR 1C 1基因的转录。在顺铂处理的2008细胞中观察到AKR 1C 1转录增加2倍，这些细胞是CCAAT盒依赖性的。这些结果表明，NF-γ调节AKR 1C 1在人卵巢癌、肺癌和肝癌细胞中的基础转录以及顺铂诱导的人卵巢癌细胞中的转录。
Dihydrodiol dehydrogenases are a family of aldo-keto reductases (AKR1Cs) involved in the metabolism of steroid hormones and xenobiotics. Herein, we have cloned and characterized the proximal promoter region of the human AKR1C1 gene. The 5′ flanking proximal promoter region of the AKR1C1 gene consists of a TATA box and an inverted CCAAT binding site. Deletion analysis of the 5′ flanking, ∼3.0kb region of the human AKR1C1 gene identified the region between −128 and −88 as the minimal proximal promoter essential for basal transcription of AKR1C1 in human ovarian (2008 and 2008/C13*), lung (H23 and A549) and liver carcinoma (HepG2) cells. Site-directed mutagenesis studies indicated that the transcription factor binding sites for NF-Y/CEBP were involved in controlling the basal transcription of AKR1C1 in all the cancer cells studied. Electrophoretic mobility shift (EMSAs) and gel-supershift assays demonstrated that the transcription factor NF-Y preferentially binds to the inverted CCAAT box at−109ATTGG−105of the AKR1C1 gene. Chromatin immunoprecipitation (ChIP) analysis confirmed the in vivo association between NF-Y and human AKR1C1 gene promoter in human ovarian, lung and liver carcinoma cells. Ectopic expression of NF-Ys increased the AKR1C1 gene transcription, whereas expression of a dominant-negative NF-YA or suppression of NF-YA decreased the AKR1C1 gene transcription. A 2-fold increase in AKR1C1 transcription was observed specifically in cisplatin-treated 2008 cells that were CCAAT box-dependent. These results indicate that the NF-Y regulates the basal transcription of AKR1C1 in human ovarian, lung and liver carcinoma cells and the cisplatin-induced transcription in human ovarian carcinoma cells.