Expression of the prostate-specific membrane antigen.
Expression of the prostate-specific membrane antigen.
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DOI：
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发表时间：
1994-04
期刊：
影响因子：
11.2
通讯作者：
R. Israeli;C. T. Powell;J. Corr;W. Fair;W. Heston
中科院分区：
文献类型：
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作者：
R. Israeli;C. T. Powell;J. Corr;W. Fair;W. Heston
我们最近克隆了一个2.65-脱氢酶互补DNA（cDNA）编码的前列腺特异性膜抗原（PSM）识别的7 E11-C5.3抗前列腺单克隆抗体。使用7 E11-C5.3抗体对LNCaP、DU-145和PC-3前列腺癌细胞系进行PSM表达的免疫组织化学分析揭示了LNCaP细胞中的强染色，而DU-145和PC-3细胞中均未检测到表达。2.65-腺苷三磷酸酶全长PSM cDNA的体外转录/翻译偶联产生与PSM的预测多肽分子量对应的M（r）84，000蛋白。用犬胰腺微粒体对该蛋白进行翻译后修饰，产生预期的M（r）100，000 PSM抗原。在真核表达载体中用全长PSM cDNA转染PC-3细胞后，我们使用7 E11-C5.3单克隆抗体通过Western分析检测PSM糖蛋白的表达。核糖核酸酶保护分析表明，PSM mRNA的表达几乎完全是前列腺特异性在人体组织中。PSM表达似乎在剥夺前列腺激素的状态下最高，并且通过类固醇激素进行神经调节，5-α-二氢睾酮将人前列腺癌细胞系LNCaP中的PSM表达下调8-10倍，睾酮将PSM下调3-4倍，并且皮质类固醇激素没有显示出显著作用。正常和恶性前列腺组织始终显示高PSM表达，而我们已经注意到异质性，有时缺乏PSM在良性前列腺增生的表达。LNCaP肿瘤原位和s.c.在裸鼠体内大量表达PSM，为研究PSM的表达调控提供了良好的体内模型系统。
We have recently cloned a 2.65-kilobase complementary DNA (cDNA) encoding the prostate-specific membrane antigen (PSM) recognized by the 7E11-C5.3 anti-prostate monoclonal antibody. Immunohistochemical analysis of the LNCaP, DU-145, and PC-3 prostate cancer cell lines for PSM expression using the 7E11-C5.3 antibody reveals intense staining in the LNCaP cells with no detectable expression in both the DU-145 and PC-3 cells. Coupled in vitro transcription/translation of the 2.65-kilobase full-length PSM cDNA yields an M(r) 84,000 protein corresponding to the predicted polypeptide molecular weight of PSM. Posttranslational modification of this protein with pancreatic canine microsomes yields the expected M(r) 100,000 PSM antigen. Following transfection of PC-3 cells with the full-length PSM cDNA in a eukaryotic expression vector, we detect expression of the PSM glycoprotein by Western analysis using the 7E11-C5.3 monoclonal antibody. Ribonuclease protection analysis demonstrates that the expression of PSM mRNA is almost entirely prostate specific in human tissues. PSM expression appears to be highest in hormone-deprived states and is hormonally modulated by steroids, with 5-alpha-dihydrotestosterone down-regulating PSM expression in the human prostate cancer cell line LNCaP by 8-10-fold, testosterone down-regulating PSM by 3-4-fold, and corticosteroids showing no significant effect. Normal and malignant prostatic tissues consistently show high PSM expression, whereas we have noted heterogeneous, and at times absent, expression of PSM in benign prostatic hyperplasia. LNCaP tumors implanted and grown both orthotopically and s.c. in nude mice abundantly express PSM, providing an excellent in vivo model system to study the regulation and modulation of PSM expression.