Human prostate-specific transglutaminase gene: Promoter cloning, tissue-specific expression, and down-regulation in metastatic prostate cancer
Human prostate-specific transglutaminase gene: Promoter cloning, tissue-specific expression, and down-regulation in metastatic prostate cancer
复制标题
DOI：
10.1016/s0090-4295(99)00298-8
复制
发表时间：
1999-12-01
期刊：
影响因子：
2.1
通讯作者：
Veltri, RW
中科院分区：
文献类型：
--
作者：
An, G;Meka, CSR;Veltri, RW
目标。目的研究人前列腺特异性谷氨酰胺转氨酶(PTGase)基因在转移性前列腺癌(CAP)中的组织特异性和差异性表达，探讨该基因在前列腺癌中的调控机制。方法采用Northern印迹杂交和聚合酶链式反应(PCR)方法，对转移性前列腺癌(CAP)组织中的组织特异性表达进行分析。采用相对定量逆转录聚合酶链式反应(RT-PCR)检测该基因在正常前列腺、前列腺癌和转移性前列腺癌中的表达差异。通过基因组文库筛选和测序，克隆了pTGase基因启动子。通过基因转染实验和氯霉素乙酰转移酶(CAT)活性检测，研究该基因的调控作用。Northern杂交和RT-PCR证实该基因仅在前列腺中表达，相对定量RT-PCR显示pTGase基因在CAP和Gleason分级较高的男性中表达缺失。在不同部位的转移性帽状组织中，86%的样本失去了该基因的表达。我们克隆并测定了pTGase基因的一个1.4kb的启动子区域。转基因和CAT分析结果支持这样的理论，即：-1到-520区域的某些元件足以指导该基因的前列腺特异性表达。-520到-1400区域的其他元件也可能对其前列腺特异性表达起作用。我们的研究结果表明，人pTGase基因只在前列腺组织中表达，而它在大多数转移性前列腺癌中的表达受到抑制。该基因的前列腺特异性表达受启动子区域的元件控制。观察到的pTGase基因在转移帽中表达的优先缺失可能在该病的发病机制和进展中起重要作用。(C)1999，爱思唯尔科学公司。
Objectives. To investigate the tissue-specific and differential expression of the human prostate-specific transglutaminase (pTGase) gene in metastatic prostate cancer (CaP) and to study how this gene is regulated in the prostate.Methods, Northern blot hybridization and polymerase chain reaction (PCR) were performed using RNA from a variety of organs to confirm prostate-specific expression of the gene. Relative quantitative reverse transcriptase-PCR (RT-PCR) was performed to investigate the differential expression of the gene among normal prostates and prostates with CaP and metastatic CaP. The pTGase gene promoter was cloned using genomic library screening and sequencing. Transfection experiments and chloramphenicol acetyltransferase (CAT) assays were performed to study the regulation of the gene.Results. Northern hybridization and RT-PCR confirmed that the gene is only expressed in the prostate, Relative quantitative RT-PCR demonstrated a loss of expression of the pTGase gene among men with CaP and higher Gleason grades. In metastatic CaP tissue from;various: sites, 86% of the samples lost expression of the gene. We cloned and sequenced a 1.4-kilobase promoter region of the pTGase gene. Transfection and CAT assay results supported the theory that certain elements in:the -1 to -520 region are sufficient to direct prostate-specific expression of the gene. Additional elements in the -520 to -1400 region may also Contribute to its prostate-specific expression.Conclusions. The results of our study demonstrate that the human pTGase gene is only expressed in prostate tissue and that its expression is inhibited in most metastatic CaP. Prostate-specific expression of the gene is controlled by elements in the promoter region. The observed preferential loss of pTGase gene expression in metastatic CaP may be important to the :pathogenesis and progression of this disease. (C) 1999, Elsevier Science Inc.