Cloning and characterization of the prostate-specific membrane antigen promoter
Cloning and characterization of the prostate-specific membrane antigen promoter
复制标题
DOI：
10.1002/(sici)1097-4644(19990901)74:3
复制
发表时间：
1999-09-01
影响因子：
4
通讯作者：
Grimes, SR
中科院分区：
文献类型：
--
作者：
Good, D;Schwarzenberger, P;Grimes, SR
前列腺特异性膜抗原（PSMA）是一种主要在正常前列腺上皮细胞和大多数前列腺癌（Cap）以及几乎所有Cap转移瘤中表达的蛋白质。在这篇文章中，我们描述了一个2-kb的人基因组DNA片段的克隆，含有5'上游非翻译区的PSMA基因，并提出证据表明，它提供启动子活性。当将DNA片段克隆到瞬时表达载体中以检查启动子活性时，在瞬时转染测定中，载体在促进几种前列腺和非前列腺细胞系中的表达中起作用。如通过缺失诱变所确定的，源自2-kb片段的3'端的614-bp片段可以代表最小PSMA启动子。当使用前列腺衍生的细胞系（DU 145和LNCaP）时，与614-bp片段相比，2-kb片段提供了更高的表达水平。在非前列腺细胞系中，使用2-kb片段增加的转录没有那么大。用232-bp启动子片段观察到很少或没有超过基础水平的转录。当在生长培养基中耗尽或补充双氢睾酮浓度时，在前列腺细胞系LNCaP细胞中未观察到对PSMA促进的瞬时表达的显著影响。出版于1999年威利-利斯公司匕首。
Prostate-specific membrane antigen (PSMA) is-a protein that is expressed predominantly in normal prostate epithelial cells and in most adenocarcinomas of the prostate (Cap) and in virtually all Cap metastases. In this article we describe the cloning or a 2-kb human genomic DNA fragment containing the 5' upstream untranslated region of the PSMA gene and present evidence that it provides promoter activity. When the DNA fragment was cloned into transient expression vectors to examine promoter activity, the vectors were functional in promoting expression in several prostate and nonprostate cell lines in transient transfection assays. A 614-bp fragment derived from the 3' end of the 2-kb fragment may represent the minimal PSMA promoter as determined by deletion mutagenesis. The 2-kb fragment compared with the 614-bp fragment provided higher expression levels when using prostate-derived cell lines (DU 145 and LNCaP). the increased transcription using the 2-kb fragment was not as great in non-prostate cell lines. Little or no transcription over basal levels was seen with a 232-bp promoter fragment. When the concentration of dihydrotestosterone was depleted or supplemented in the growth medium, no significant effect was seen on PSMA-promoted transient expression in LNCaP cells, a prostate cell line. Published 1999 Wiley-Liss, Inc.dagger.