ANDROGEN INDUCTION OF A HUMAN PROSTATE-SPECIFIC KALLIKREIN, HKLK2 - CHARACTERIZATION OF AN ANDROGEN RESPONSE ELEMENT IN THE 5' PROMOTER REGION OF THE GENE
ANDROGEN INDUCTION OF A HUMAN PROSTATE-SPECIFIC KALLIKREIN, HKLK2 - CHARACTERIZATION OF AN ANDROGEN RESPONSE ELEMENT IN THE 5' PROMOTER REGION OF THE GENE
复制标题
DOI：
10.1021/bi00076a020
复制
发表时间：
1993-06-29
期刊：
影响因子：
2.9
通讯作者：
YOUNG, CYF
中科院分区：
文献类型：
--
作者：
MURTHA, P;TINDALL, DJ;YOUNG, CYF
人前列腺特异性激肽释放酶、人腺激肽释放酶-1 (hKLK2) 和前列腺特异性抗原 (hKLK3) 已被证明受雄激素调节。为了确定这些基因的雄激素诱导是否通过雄激素反应元件进行转录调节，将 hKLK2 启动子 DNA 片段连接到无启动子的氯霉素乙酰转移酶 (CAT) 报告基因，并在无雄激素受体的人前列腺细胞系 PC-3 中与雄激素受体表达载体共转染。剂量反应和类固醇特异性实验表明，hKLK2 启动子以配体特异性方式赋予雄激素受体介导的基因诱导作用。此外，hKLK2启动子DNA的5'缺失构建体和缺乏推定的雄激素反应元件(ARE)的5'半位点的内部缺失构建体用于表明推定的ARE确实在前列腺细胞中充当功能性ARE。此外，来自 hKLK2 和 hKLK3 的多个 ARE 能够重建雄激素诱导，进一步强化了 ARE 具有功能性的论点。尽管之前的研究表明hKLK3 mRNA的表达水平高于hKLK2，但我们的结果表明hKLK2 ARE可能比hKLK3 ARE具有更高的雄激素诱导能力。这些结果表明其他顺式作用元件可能参与协调hKLK2和hKLK3基因的体内雄激素诱导。
The human prostate-specific kallikreins, human glandular kallikrein-1 (hKLK2) and prostate-specific antigen (hKLK3), have been shown to be regulated by androgens. To determine whether the androgen induction of these genes is transcriptionally regulated via an androgen response element, an hKLK2 promoter DNA fragment was linked to a promoterless chloramphenicol acetyltransferase (CAT) reporter gene and cotransfected with an androgen receptor expression vector in an androgen receptor-less human prostate cell line, PC-3. Dose response and steroid specificity experiments showed that the hKLK2 promoter confers androgen receptor-mediated gene induction in a ligand-specific manner. Moreover, 5' deletion constructs of the hKLK2 promoter DNA and internal deletion constructs devoid of the 5' half-site of the putative androgen responsive element (ARE) were used to show that the putative ARE is indeed acting as a functional ARE in prostate cells. In addition, multiple AREs from both hKLK2 and hKLK3 were able to reconstitute androgenic induction, further strengthening the argument that the AREs are functional. Although previous studies have shown that hKLK3 mRNA is expressed at a higher level than that of hKLK2, our results suggest that the hKLK2 ARE may have higher androgenic inducibility than the hKLK3 ARE. These results suggest that other cis-acting elements may be involved in coordinating in vivo androgenic induction of hKLK2 and hKLK3 genes.