膀胱癌组织中miRNA-720、miRNA-191差异表达的意义

目的 探讨miRNA-720、miRNA-191在膀胱癌中差异表达的意义。方法 取手术切除的24例新鲜膀胱癌组织（T1~4）及12例前列腺增生患者膀胱组织保存于液氮中,Trizol试剂提取总RNA,NanoDrop 2000及变性琼脂糖凝胶电泳进行RNA检测,用标记酶Hy3TM荧光基团标记RNA的探针和miRCURYTM芯片杂交,GenePix 4000B芯片扫描仪及GenePix pro V6.0进行图像采集和数据分析,对差异表达的miRNA进行检测,实时定量逆转录PCR（qRT-PCR）对部分差异表达的miRNA进行验证。结果 与正常膀胱黏膜组织比较,非浸润性膀胱癌组织中有77个miRNA表达增加,72个表达降低;浸润性膀胱癌组织中有42个miRNA表达增加,104个表达降低。qRT-PCR检测证实miRNA-720和miRNA-191的表达水平与基因芯片结果一致。结论 非浸润性和浸润性膀胱癌组织中miRNA的差异表达普遍存在,miRNA-720可以作为膀胱癌检测的一个组织标志物,而miRNA-191在浸润性膀胱癌中的表达水平较非浸润性膀胱癌低（P〈0.05）,可作为膀胱癌是否浸润肌层的一个组织标志物。

Differential expression and significance of miRNA-720, miRNA-191 in bladder cancer

Objective To identify the differentially expressed microRNA （miRNA） 720, miR- NA-191 in bladder cancer compared with normal bladder. Methods 24 fresh tissues of bladder cancer and 12 normal bladder mucosa were stored in liquid nitrogen. Total RNA was extracted by using Trizol reagents, purified and tested by denaturing agarose gel electrophoresis and NanoDrop 2000. The purified RNA were labeled by Hy3TM fluorescent label then hybridized with miRCURYTM Array. MiRNA arrays＇ scanning and analysis were performed with the Axon GenePix 4000B microarray scanner and GenePix proV6.0. The scanned result was validated by using qRT-PCR. Re- sults 77 miRNAs expression was increased and 72 miRNAs expression was decreased in non-inva- sire bladder cancer compared to the normal miRNAs, 42 miRNAs expression was increased and 104 miRNAs expression was decreased in invasive bladder cancer compared to the normal miRNA. The expression levels of miRNA-720 and miRNA-191 were verified by qRT-PCR. Conclusions Differently expressed level of miRNAs were detected between muscle invasive bladder cancer and non-muscle invasive bladder cancer, and we found that miRNA-720 might be a marker for bladder cancer diagnosis. The express of miRNA-191 in muscle invasive bladder cancer is lower compared with those with non-muscle invasive, indecating that miRNA-191 might be a diagnositic marker for bladder cancer with muscle invasive.