Coutinho-Camillo Cláudia M, Salaorni Sibeli, Sarkis Alvaro S, Nagai Maria A
Laboratório de Genética Molecular do Câncer, Disciplina de Oncologia, Departamento de Radiologia, FMUSP, São Paulo, Brazil.
Cancer Genet Cytogenet. 2006 Apr 15;166(2):130-8. doi: 10.1016/j.cancergencyto.2005.09.012.
Androgens play an important role in growth and maintenance of prostate cells. The actions of androgens are mediated by the androgen receptor (AR), a transcription factor member of the super-family of nuclear hormone receptors. Androgen regulated genes (ARGs) are potential markers for early diagnosis and treatment of prostate cancer patients. In the present study, we used DDRT-PCR (differential display reverse transcriptase polymerase chain reaction) technique in order to investigate differentially expressed genes in the prostate cancer cell line LNCaP after treatment with dihydrotestosterone and bicalutamide for 6, 24, and 48 hours. Fifty-five differentially expressed fragments were isolated, cloned, and sequenced. Sequencing analysis of these fragments revealed 56 different transcripts that showed homology to transcription factors, cell cycle regulators, metabolic enzymes, and hypothetical proteins. Among the differentially expressed genes, SPA17 and DDEF2 were further validated using quantitative real time RT-PCR (qPCR) in a series of 25 prostate tumor samples. The DDEF2 gene is involved in adhesion and cell migration of monocytes, and the SPA17 gene might be involved in cellular signal transduction. The transcripts of both, SPA17 and DDEF2 genes, showed altered pattern of expression in the group of prostate tumors analyzed by qPCR. The differentially expressed genes identified in this study might provide new insights into the androgen signaling pathways in prostate cells.
雄激素在前列腺细胞的生长和维持中起着重要作用。雄激素的作用由雄激素受体（AR）介导，AR是核激素受体超家族的转录因子成员。雄激素调节基因（ARGs）是前列腺癌患者早期诊断和治疗的潜在标志物。在本研究中，我们使用差异显示逆转录聚合酶链反应（DDRT-PCR）技术，以研究在二氢睾酮和比卡鲁胺处理6、24和48小时后，前列腺癌细胞系LNCaP中差异表达的基因。分离、克隆并测序了55个差异表达片段。对这些片段的测序分析揭示了56种不同的转录本，它们与转录因子、细胞周期调节因子、代谢酶和假定蛋白具有同源性。在差异表达的基因中，使用定量实时RT-PCR（qPCR）在一系列25个前列腺肿瘤样本中进一步验证了SPA17和DDEF2。DDEF2基因参与单核细胞的黏附和细胞迁移，SPA17基因可能参与细胞信号转导。通过qPCR分析，SPA17和DDEF2基因的转录本在前列腺肿瘤组中均显示出改变的表达模式。本研究中鉴定出的差异表达基因可能为前列腺细胞中的雄激素信号通路提供新的见解。
