与人PC-1同源的鼠PC-1基因的启动子的分子克隆和特性分析

为了鉴定鼠mPC-1基因表达的调控元件，克隆并分析了该基因的启动子.构建了一系列mPC-1基因启动子的截短序列.通过荧光素酶报道基因,分析了它们在前列腺癌细胞和其它细胞中的表达.结果表明,在AR阳性细胞系中,mPC-1基因启动子活性远远高于SV40和p61-PSA 启动子，mPC-1基因启动子 599 bp 至449bp 可能含有一个负调控元件； mPC-1 1.1 kb 启动子控制的表达主要在前列腺癌细胞系中； 雄激素可调控mPC-1 1.1kb 启动子表达.mPC-1 1.1kb 序列是一个有前列腺癌细胞特异性和较强的启动子，经过进一步的修饰有可能作为一种有用的前列腺癌基因治疗元件.

Molecular Cloning and Characterization of a Putative Promoter Region of the mPC-1 Gene Homologous to hPC-1

To identify the regulatory region that are responsible for the expression of mPC-1,we have isolated and characterized the mPC-1 gene promoter.Sequence analysis of the mPC-1 5'-flanking region and a series of truncated constructs were performed,which were transiently transfected into the prostate cancer cell lines and non-prostate cancer cell lines and analyzed through Dual-luciferase reporter assay system.The relative activity of mPC-1 gene promoter was by far higher than pGL3-control containing SV40 promoter and enhancer and p61-PSA containing hPSA 6 kb promoter in AR(androgen receptor,AR)-positive prostate cancer cell lines.The region from 599 bp to 449 bp of mPC-1 promoter might contain a negative regulatory element.The expression of mPC-1 1.1 kb fragment is mainly restricted into prostate cancer cell lines.The relative activity of mPC-1 1.1 kb 5'-flanking region was regulated by androgen.The results demonstrated that the 1.1 kb fragment of mPC-1 5'-flanking region was relatively strong and prostate cancer cell specific promoter region.The 1.1 kb promoter of mPC-1 gene might be well suited to prostate cancer gene therapy if the promoter was properly modified.