Multiple androgen response elements cooperate in androgen regulated activity of the type 1 neutral endopeptidase promoter.
Multiple androgen response elements cooperate in androgen regulated activity of the type 1 neutral endopeptidase promoter.
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多个雄激素反应元件在 1 型中性内肽酶启动子的雄激素调节活性中协同作用。
DOI：
10.1016/j.mce.2006.07.004
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发表时间：
2006
影响因子：
4.1
通讯作者：
Nanus,DavidM
中科院分区：
文献类型：
--
作者：
Zheng,Rong;Shen,Ruoqian;GoodmanJr,OscarB;Nanus,DavidM
中性内肽酶（NEP）基因在前列腺癌细胞中受雄激素的转录调控。我们先前在NEP基因中鉴定了一个雄激素应答元件（NEP-ARE）和一个雄激素应答区（NEP-ARR），它们一起仅传递中度雄激素诱导性[Mol. Cell.年. 170（2000）131]。因此，我们对NEP基因的整个基因组结构进行了表征，并鉴定了ARE 1（ACTCAACAtttgTGTCCTTT）和ARE 2（CAGGACAtttTGTCCC），它们分别位于3′-非翻译区和内含子17中。通过将含有三个拷贝的ARE 1或ARE 2的pGL-3-荧光素酶报告质粒转染到PC-3细胞中来测定转录的类固醇依赖性增强。雄激素（AR）使荧光素酶活性增加3.6倍（ARE 1）和5倍（ARE 2），地塞米松使荧光素酶活性增加4.2倍（ARE 1）和8.2倍（ARE 2），孕酮使荧光素酶活性增加3倍（ARE 1）和4.1倍（ARE 2）。ARE 1和ARE 2序列的突变完全废除雄激素诱导。我们接下来表明，ARE 1和ARE 2都参与NEP基因的转录调控，证明在体外和体内与AR结合，如通过电泳迁移率凝胶位移和染色质免疫沉淀（ChIP）测定所确定的。此外，ARE 1和ARE 2在SV 40启动子和天然NEP 1型启动子中介导协调的雄激素诱导。这些数据表明新鉴定的ARE 1和ARE 2与先前鉴定的NEP-ARE一起作为雄激素反应元件起作用，并且NEP基因的雄激素调节由前列腺癌细胞中多个战神的协调作用调节。
The neutral endopeptidase (NEP) gene is transcriptionally regulated by androgen in prostate cancer cells. We previously identified in the NEP gene an androgen responsive element (NEP-ARE) and an androgen responsive region (NEP-ARR) that together conveyed only moderate androgen-inducibility [Mol. Cell. Endocrinol. 170 (2000) 131]. Therefore, we characterized the entire genomic structure of the NEP gene and identified ARE1 (ACTCAACAttgTGTCCTTT) and ARE2 (CAGGACAtttTGTCCC), which are located in the 3′-untranslated region and in intron 17, respectively. Steroid-dependent enhancement of transcription was assayed by transfecting the pGL-3-luciferase reporter plasmid containing three copies of ARE1 or ARE2 into PC-3 cells. Luciferase activities were increased 3.6-fold (ARE1) and 5-fold (ARE2) by androgen (AR), 4.2-fold (ARE1) and 8.2-fold (ARE2) by dexamethasone, and 3-fold (ARE1) and 4.1-fold (ARE2) by progesterone. Mutation of the ARE1 and ARE2 sequences completely abrogated androgen-inducibility. We next showed that both ARE1 and ARE2 are involved in the transcriptional regulation of the NEP gene, demonstrating in vitro and in vivo binding with AR as determined by electrophoretic mobility gel shift and chromatin immunoprecipitation (ChIP) assays, Furthermore, ARE1 and ARE2 mediate coordinated androgen-inducibility in both an SV40 promoter and the native NEP type 1 promoter. These data indicate the newly identified ARE1 and ARE2 together with the previously identified NEP-ARE function as androgen response elements, and that androgen regulation of the NEP gene is regulated by the coordinated action of multiple AREs in prostate cancer cells.