摘要
目的筛选和鉴定新的前列腺癌相关基因,探讨其在前列腺癌中的表达情况,为研究其在前列腺癌发生、发展中的作用奠定基础。方法通过生物学信息筛选,RT-PCR验证,确定前列腺癌相关新基因PCAG1,然后提取14例配对前列腺癌及癌旁组织标本中总RNA,经逆转录获得cDNA,应用RT-PCR检测标本中PCAG1 mRNA表达水平。免疫组织化学染色检测PCAG1在38例前列腺癌组织及配对邻近正常组织中蛋白表达水平,免疫荧光法确定PCAG1蛋白亚细胞定位,并通过统计学方法分析PCAG1表达水平与前列腺癌的关系。结果 RT-PCR及免疫组化显示PCAG1在前列腺癌组织中的表达明显高于癌旁正常组织,且只在少数几种正常组织中低表达,多数正常组织中无表达,呈现明显的前列腺癌特异高表达的特性;免疫荧光确定PCAG1蛋白主要定位于线粒体中。结论 PCAG1在前列腺癌组织中高表达,提示可能与前列腺癌的发生、发展相关。PCAG1独特的转录调控模式预示其具有潜在的临床应用价值。
Objective To screen and identify a new prostate cancer-associated gene, analyzing its expressicn pattern,and evaluate its expression in prostate cancer for further study of its role in the oncogmesis and development of prostate cancer. Methods Candidate genes were obtained with combined bioinformatics screening. Prostate cancer-associated gene 1 （PCAG1） was then identified by performing reverse transcription （RT）-PCR to validate prostate cancer specificity. Total RNA of 1,t paired prostate cancer tissues and adjacent normal tissues were isolated and reverse transcription was performed to obtain the eDNA. The expression of PCAG1 mRNA was detected by RT-PCR, and lhe expression of PCAG1 protein was evaluated by immunohistochemical essay . Immunoflures- cent staining was used to discover the subccllular location of PCAG1 protein and statistic analysis was performed to explore the relationship between expression of PCAG1 and prostate cancer. Re- sults PCAG1 mRNA was highly expressed in prostate cancer tissues while being almost absent in all tested common normal tissues and paired adjacent normal prostate tissues. Furthermore, PCAG1 protein was also highly expressed in prostate cancer tissues while few common normal tissues, other common malignant tumors and paired adjacent normal prostate tissues showed even low levels of ex- pression. The result of immunoflurescent staning indicated that PCAG1 protein was located in the mitochondria. Conclusions PCAG1 is highly expressed in prostate cancer tissues, suggesting it may be involved in the oncogenesis and development of prostate cancer. The unique expression pat- tern of PCAG1 suggests its potential in certain clinical applications.
出处
《现代泌尿生殖肿瘤杂志》
2013年第3期155-159,共5页
Journal of Contemporary Urologic and Reproductive Oncology
基金
国家自然科学基金(81101922)
广东省医学科研基金(A2012584
A2013606)
2013年深圳市科技研发资金知识创新计划基础研究项目(JCYJ20130402114702124)
