膀胱尿路上皮癌(Ⅰ级)miRNAs差异表达及意义  

Study of differentially expressed microRNAs in urothelial carcinoma of the bladder(Ⅰgrade) and its significance

作　　者：徐锋[1] 高建平[1] 张征宇[1] 葛京平[1] 程文[1] 位志峰[1] 

机构地区：[1]南京大学医学院临床学院南京军区南京总医院泌尿外科,江苏南京210002

出　　处：《中国老年学杂志》2011年第5期793-797,共5页Chinese Journal of Gerontology

基　　金：国家自然科学基金资助项目(30772278);南京军区南京总医院院内青年课题(2010-2011)

摘　　要：目的研究膀胱尿路上皮癌miRNAs的差异表达。方法取手术切除新鲜膀胱尿路上皮癌及正常膀胱黏膜组织各3例保存于液氮中;Trizol试剂提取总RNA;使用Nanodrop及变性琼脂糖凝胶电泳进行RNA质量检测;用标记酶Hy3TM荧光基团标记RNA的探针和miRCURYTM芯片杂交;使用GenePix4000B芯片扫描仪及GenePix pro V6.0完成图像采集和数据分析;荧光定量RT-PCR方法验证。结果膀胱尿路上皮癌与正常膀胱黏膜上皮组织共有192个差异表达基因,其中71个上调,124个下调;差异表达miRNAs经随机RT-PCR验证而证实。结论膀胱尿路上皮癌与正常膀胱黏膜组织中存在差异miRNAs基因,这种miRNAs差异可能导致膀胱尿路上皮癌的发生及发展。Objective To investigate the gene expression disparity and the significance of urothelial carcinoma of the bladder（BUC）.Methods The fresh tissues of BUC were taken and normal bladder mucosa were stored in liquid nitrogen container,each 3 cases.Total RNA was extracted by Trizol reagents.RNA quantification and quality control were tested by denaturing agarose gel electrophoresis and NanoDrop ND-1000.MiRNA probes were labelled by Hy3TM fluorescent label,then hybridized with miRCURYTM Array.MiRNA arrays scanning and analysis were performed by Axon GenePix 4000B microarray scanner and GenePix pro V6.0.The scanned result was additionally validated by RT-PCR.Results 195 genes were differentially expressed in BUC,including 71 genes up-regulated,and 124 down-regulated.The result of miRNA arrays was consistent with real time RT-PCR.Conclusions Genes are differentially expressed between BUC and normal bladder mucosa,which may participate in the pathogenesis and development of BUC.

关 键 词：MIRNAS 膀胱尿路上皮癌 基因表达谱 差异性 

分 类 号：R737.14[医药卫生—肿瘤]