A small composite probasin promoter confers high levels of prostate-specific gene expression through regulation by androgens and glucocorticoids in vitro and in vivo
A small composite probasin promoter confers high levels of prostate-specific gene expression through regulation by androgens and glucocorticoids in vitro and in vivo
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DOI：
10.1210/en.141.12.4698
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发表时间：
2000-12-01
期刊：
影响因子：
4.8
通讯作者：
Matusik, RJ
中科院分区：
文献类型：
--
作者：
Zhang, JF;Thomas, TZ;Matusik, RJ
瞬时转染研究表明，probasin（PB）启动子赋予雄激素的选择性超过其他类固醇激素，转基因动物研究表明，PB启动子将靶向雄激素，但不是糖皮质激素，调节前列腺特异性的方式。以前的PB启动子要么靶向低水平的转基因表达，要么变得太大而不能方便地使用。目的是设计一个小的PB启动子，设定高水平的前列腺特异性转基因表达。因此，在前列腺和非前列腺细胞系和转基因小鼠模型中产生并测试了与细菌氯霉素乙酰转移酶报告基因（ARR（2）PBCAT）偶联的复合probasin启动子（ARR（2）PB）。在PC-3、LNCaP和DU 145前列腺癌细胞系中，ARR（2）PB启动子提供基础表达，并在与各自的类固醇受体共转染后对雄激素和糖皮质激素处理产生诱导。ARR（2）PBCAT在非前列腺COS-1、MCF-7、ZR-75 - 1和PANC-1细胞系中的基础表达非常低;然而，当细胞与AR或GR共转染时，CAT活性可被雄激素和糖皮质激素诱导。与转染研究相反，ARR（2）PBCAT转基因表达在转基因小鼠中对前列腺上皮仍具有高度特异性。CAT活性下降去势后，可以诱导雄激素，此外，糖皮质激素。这表明靶向前列腺特异性上皮表达所需的必要序列包含在复合ARR（2）PB最小启动子中，并且高转基因表达现在可以由雄激素和糖皮质激素调节。ARR2PB启动子代表一种新型糖皮质激素诱导型启动子，其可用于产生转基因小鼠模型和用于治疗人类前列腺癌的病毒基因治疗载体。
Transient transfection studies have shown that the probasin (PB) promoter confers androgen selectivity over other steroid hormones, and transgenic animal studies have demonstrated that the PB promoter will target androgen, but not glucocorticoid, regulation in a prostate-specific manner. Previous PB promoters either targeted low levels of transgene expression or became too large to be conveniently used. The goal was to design a PB promoter that would be small, Set target high levels of prostate-specific transgene expression. Thus, a composite probasin promoter (ARR(2)PB) coupled to the bacterial chloramphenicol acetyltransferase reporter (ARR(2)PBCAT) was generated and tested in prostatic and nonprostatic cell lines and in a transgenic mouse model. In PC-3, LNCaP, and DU145 prostate cancer cell lines, the ARR(2)PB promoter gave basal expression and was induced in response to androgen and glucocorticoid treatment after cotransfection with the respective steroid receptor. Basal expression of ARR(2)PBCAT in the nonprostatic COS-1, MCF-7, ZR-75-1, and PANC-1 cell lines was very low; however, CAT activity could be induced in response to androgens and glucocorticoids when cells were cotransfected with either the AR or GR. In contrast to the transfection studies, ARR(2)PBCAT transgene expression remained highly specific for prostatic epithelium in transgenic mice. CAT activity decreased after castration, and could be induced by androgens and, in addition, glucocorticoids. This demonstrates that the necessary sequences required to target prostate-specific epithelial expression are contained within the composite ARR(2)PB minimal promoter, and that high transgene expression can now be regulated by both androgens and glucocorticoids. The ARR2PB promoter represents a novel glucocorticoid inducible promoter that can be used for the generation of transgenic mouse models and in viral gene therapy vectors for the treatment of prostate cancer in humans.