TRANSCRIPTIONAL REGULATION OF ANDROGEN RECEPTOR GENE-EXPRESSION IN SERTOLI CELLS AND OTHER CELL-TYPES
TRANSCRIPTIONAL REGULATION OF ANDROGEN RECEPTOR GENE-EXPRESSION IN SERTOLI CELLS AND OTHER CELL-TYPES
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DOI：
10.1016/0303-7207(92)90020-7
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发表时间：
1992-10-01
影响因子：
4.1
通讯作者：
GROOTEGOED, JA
中科院分区：
文献类型：
--
作者：
BLOK, LJ;THEMMEN, APN;GROOTEGOED, JA
雄激素受体（AR）mRNA表达的调节进行了研究，支持细胞和管周肌样细胞分离自未成熟的大鼠睾丸，并在淋巴结癌细胞系来自人前列腺（LNCaP）。加入二丁酰环腺苷酸（dbcAMP）的Sertoli细胞培养物中的AR mRNA的表达（5小时），这是随后的AR mRNA的表达（24-72小时）逐渐增加的快速瞬时下降。dbcAMP的这种作用模拟了促卵泡激素（FSH）的作用。在管周肌样细胞中，只有一个温和的，但长期的下降，在孵育过程中的dbcAMP的存在下，并在LNCaP细胞中没有dbcAMP对AR mRNA的表达的影响进行了观察。当支持细胞或管周肌样细胞培养的雄激素的存在下，AR mRNA的表达在这些细胞类型没有改变。这与LNCaP细胞相反，LNCaP细胞在雄激素处理期间显示AR mRNA表达显著降低。在本实验中，还研究了Sertoli细胞和LNCaP细胞中AR基因表达的转录调控。用一系列由AR启动子驱动的荧光素酶报告基因构建体转染新鲜分离的Sertoli细胞簇。发现向转染的Sertoli细胞中添加dbcAMP导致报告基因表达的少量但一致的增加（其被解释为由AR启动子活性引起）;仅含有cDNA序列的AR 5'非翻译区的构建体不显示这种调节。当将合成雄激素R1881添加到细胞培养物中时，转染到LNCaP细胞中的相同构建体未显示任何转录下调。核转录延长实验（run-on）表明，LNCaP细胞中雄激素诱导的AR mRNA下调是由于AR基因转录受到抑制所致，提示在Sertoli细胞和LNCaP细胞中，激素对AR基因转录的影响在AR表达的调节中起作用。然而，这些细胞中的AR基因转录受到差异调节。
Regulation of androgen receptor (AR) mRNA expression was studied in Sertoli cells and peritubular myoid cells isolated from immature rat testis, and in the lymph node carcinoma cell line derived from a human prostate (LNCaP). Addition of dibutyryl-cyclic AMP (dbcAMP) to Sertoli cell cultures resulted in a rapid transient decrease in AR mRNA expression (5 h), which was followed by a gradual increase in AR mRNA expression (24-72 h). This effect of dbcAMP mimicked follicle-stimulating hormone (FSH) action. In peritubular myoid cells, there was only a moderate but prolonged decrease during incubation in the presence of dbcAMP, and in LNCaP cells no effect of dbcAMP on AR mRNA expression was observed.When Sertoli cells or peritubular myoid cells were cultured in the presence of androgens, AR mRNA expression in these cell types did not change. This is in contrast to LNCaP cells, that showed a marked reduction of AR mRNA expression during androgen treatment.In the present experiments, transcriptional regulation of AR gene expression in Sertoli cells and LNCaP cells was also examined. Freshly isolated Sertoli cell clusters were transfected with a series of luciferase reporter gene constructs, driven by the AR promoter. It was found that addition of dbcAMP to the transfected Sertoli cells resulted in a small but consistent increase in reporter gene expression (which was interpreted as resulting from AR promoter activity); a construct that only contained the AR 5' untranslated region of the cDNA sequence did not show such a regulation. The same constructs, transfected into LNCaP cells, did not show any transcriptional down-regulation when the synthetic androgen R1881 was added to the cell cultures. A nuclear transcription elongation experiment (run-on), however, demonstrated that androgen-induced AR mRNA down-regulation in LNCaP cells resulted from an inhibition of AR gene transcription.The present results indicate that in Sertoli cells and LNCaP cells, hormonal effects on AR gene transcription play a role in regulation of AR expression. However, AR gene transcription in these cells is differentially regulated.