TISSUE-SPECIFIC AND HORMONAL-REGULATION OF HUMAN PROSTATE-SPECIFIC GLANDULAR KALLIKREIN
TISSUE-SPECIFIC AND HORMONAL-REGULATION OF HUMAN PROSTATE-SPECIFIC GLANDULAR KALLIKREIN
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DOI：
10.1021/bi00118a026
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发表时间：
1992-01-28
期刊：
影响因子：
2.9
通讯作者：
TINDALL, DJ
中科院分区：
文献类型：
--
作者：
YOUNG, CYF;ANDREWS, PE;TINDALL, DJ
激肽释放酶参与许多特定多肽前体的翻译后加工。以前，人腺激肽释放酶（hGK-1）的mRNA已被确定在前列腺;然而，hGK-1蛋白尚未被确定和表征。因此，其在前列腺中的生理功能尚不清楚。在这项研究中，我们已经从人腺癌细胞系LNCaP中分离出全长hGK-1 cDNA。体外翻译实验表明，从该cDNA产生的hGK-1蛋白的分子大小与前列腺特异性抗原（PSA）的分子大小相似，PSA是一种仅在前列腺中产生的蛋白。用hGK-1特异性寡核苷酸探针（77个碱基）进行原位杂交，结果表明hGK-1 mRNA定位于前列腺上皮细胞。该探针能区分hGK-1 mRNA和PSA mRNA。hGK-1 mRNA与PSA mRNA在前列腺上皮细胞中共定位。此外，原位杂交研究显示，hGK-1 mRNA在人良性前列腺增生组织中的水平约为PSA mRNA的一半。此外，我们已经证明，hGK-1 mRNA在LNCaP细胞中受到雄激素调节。时程分析显示，mibolerone作用5 h后hGK-1 mRNA水平显著升高，9 h达高峰。此外，hGK-1的mRNA水平增加的双氢睾酮，但不是由地塞米松或己烯雌酚治疗。非代谢性抗雄激素药氟替卡松可抑制雄激素效应。这些研究表明hGK-1 mRNA的表达受雄激素通过雄激素受体调节。
Kallikreins are involved in the posttranslational processing of a number of specific polypeptide precursors. Previously, human glandular kallikrein (hGK-1) mRNA has been identified in the prostate; however, the hGK-1 protein has not been identified and characterized. Therefore, its physiologic function in the prostate is not known. In this study, we have isolated a full-length hGK-1 cDNA from a human adenocarcinoma cell line, LNCaP. In vitro translation experiments demonstrated that the molecular size of the hGK-1 protein generated from this cDNA is similar to that of prostate-specific antigen (PSA), a protein which is produced exclusively in the prostate. In situ hybridization with a hGK-1-specific oligonucleotide probe (77 bases), which can differentiate hGK-1 mRNA from PSA mRNA, demonstrated the hGK-1 mRNA to be located in the prostate epithelium. The hGK-1 mRNA was colocalized with PSA mRNA in prostatic epithelia. Moreover, in situ hybridization studies revealed that the level of hGK-1 mRNA in human benign prostatic hyperplasia tissues is approximately half that of PSA mRNA. Furthermore, we have demonstrated that hGK-1 mRNA is under androgenic regulation in LNCaP cells. Time course analysis revealed that hGK-1 mRNA levels increased significantly at 5 h of mibolerone treatment and reached maximal levels by 9 h. In addition, hGK-1 mRNA levels were increased by dihydrotestosterone, but not by dexamethasone or diethylstilbestrol treatments. Flutamide, a nonmetabolized anti-androgen, repressed the androgenic effects. These studies suggest that expression of hGK-1 mRNA is regulated by androgen via the androgen receptor.