Regulation of prostasin expression and function in the prostate
Regulation of prostasin expression and function in the prostate
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DOI：
10.1002/pros.10346
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发表时间：
2004-04-01
期刊：
影响因子：
2.8
通讯作者：
Chai, KX
中科院分区：
文献类型：
--
作者：
Chen, LM;Zhang, XC;Chai, KX
背景。侵袭抑制因子前列腺蛋白酶在前列腺癌中下调，但其机制尚不清楚。在精囊中发现了前列腺素结合蛋白（PBP），但其身份尚不清楚。采用甲基化敏感限制性内切酶基因组Southern blot分析前列腺癌细胞系中前列腺蛋白酶基因启动子区域。RT-PCR检测前列腺素在去甲基化、组蛋白去乙酰化酶抑制和神经生长因子（NGF）治疗下的表达。采用液相柱色谱法对小鼠精囊中的PBP进行纯化。通过氨基酸序列分析、重组蛋白表达、蛋白酶抑制和结合实验对PBP进行了进一步的鉴定。免疫组织化学和Western blot检测PBP在前列腺和前列腺癌细胞中的表达。在DU-145和PC-3细胞中，启动子DNA甲基化部分导致前列腺蛋白酶下调，而NGF可诱导前列腺蛋白酶表达。PBP被鉴定为蛋白酶连接蛋白1 (PN-1)，一种蛇形蛋白。PN-1抑制前列腺蛋白酶丝氨酸蛋白酶活性，在前列腺上皮细胞（PrECs）和前列腺癌细胞中表达，能够与膜锚定的前列腺蛋白酶结合。前列腺蛋白酶的表达和功能受前列腺组织环境因素的调控。(C) 2003 Wiley-Liss, Inc。
BACKGROUND. The invasion suppressor prostasin is down-regulated in prostate cancer, but the mechanism is unknown. A prostasin-binding protein (PBP) was found in the seminal vesicles, but its identity remains unclear.METHODS. Genomic Southern blot analysis using methylation sensitive restriction endonucleases was employed to examine the prostasin gene promoter region in prostate cancer cell lines. RT-PCR was employed to examine prostasin expression under demethylation, histone deacetylase inhibition, and nerve growth factor (NGF) treatment. Liquid column chromatography was employed to purify the PBP from mouse seminal vesicles. The PBP was further characterized by amino acid sequence analysis, recombinant protein expression, protease inhibition and binding assays. Immunohistochemistry and Western blot analysis were used to evaluate PBP expression in the prostate and prostate cancer cells.RESULTS. Promoter DNA methylation partly causes the prostasin down-regulation in DU-145 and PC-3 cells, while prostasin expression can be induced by NGF. The PBP is identified to be protease nexin-1 (PN-1), a serpin. PN-1 inhibits prostasin's serine protease activity, is expressed by prostate epithelial cells (PrECs) and prostate cancer cells, and capable of binding to membrane-anchored prostasin.CONCLUSIONS. Prostasin's expression and function are regulated by factors in the prostate tissue environment. (C) 2003 Wiley-Liss, Inc.