摘要
目的microRNA(miRNA)通过调节原癌基因/抑癌基因或miRNA自身丢失/突变参与肿瘤的发生与形成,其差异表达可作为肿瘤早期诊断、疗效、预后评估的指标,有望成为肿瘤治疗的新靶点。膀胱尿路上皮癌是我国泌尿系恶性肿瘤中最常见的肿瘤,其发病机制目前仍不清楚。文中对Ⅱ级膀胱尿路上皮癌细胞中miRNA的差异表达及意义进行初步探讨。方法取手术切除的新鲜Ⅱ级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,用Trizol试剂提取总RNA,用Nano-drop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,用GenePix4000B芯片扫描仪及GenePixproV6.0进行图像采集和数据分析,用荧光定量RT-PCR验证。结果Ⅱ级膀胱尿路上皮癌与正常膀胱黏膜上皮组织共有21个差异表达基因,表达上调者12个:hsa-miR-203、hsa-miR-876-3p、hsa-miR-29b-1*、hsa-miR-665、hsa-miR-668、hsa-miR-181b、hcmv-miR-US25-2-5p、hsa-miR-768-3p、hsa-miR-548d-5p、hsa-miR-518c*、hsa-miR-145*、hsa-miR-363*,表达下调者9个:hsa-miR-126、hsa-miR-140-3p、hsa-miR-223、hsa-miR-300、hsa-miR-24-2*、hsa-miR-627、hsa-miR-142-3p、hsa-miR-744、hsa-miR-142-5p随机选择hsa-miR-363*、hsa-miR-627、hsa-miR-768-3p、hsa-miR-3004个基因差异表达的miRNA用RT-PCR验证,与基因芯片结果一致。结论膀胱尿路上皮癌与正常膀胱黏膜组织中存在差异表达的miRNA基因,可能是导致膀胱尿路上皮癌的发生及发展的重要原因。
Objective Recent studies indicate that microRNAs, by regulating the proto-oncogenes or tumor suppressor genes, or by their loss or mutation, are involved in tumor formation. The difference of microRNA expression can be used as indicators of early tumor diagnosis, treating efficacy and prognosis. The microRNAs can be new targets of tumor treatment. Urothelial carcinoma of the bladder is the most common malignancy of urinary system in China, its pathogenesis has not been clarified yet. This study is to investigate the microRNA expression disparity and its significance in urothelial carcinoma of the bladder （BUC）. Methods Fresh tissues of BUC and normal bladder mucosa （3 cases each） were stored in liquid nitrogen. Total RNA was extracted by using Trizol reagents, purified and tested by denaturing agarose gel electrophoresis and NanoDrop ND -1000. The purified RNA were labeled by Hy3TM fluorescent label, then hybridized with miRCURYTM Array. MicroRNA arrays＇scanning and analysis were performed with the Axon GenePix 4000B microarray scanner and GenePix pro V6.0. The scanned result was validated by using RT-PCR. Results Twenty-one genes were differentially expressed in BUC,including 12 genes up-regulated（hsa-miR-203, hsa-miR-876-3p, hsa-miR-29b-1 ＊, hsa-miR-665, hsa-miR-668, hsa-miR-181b, hcmv-miR-US25-2-5p, hsa-miR-768-3p, hsa-miR-548d-5p, hsa-miR-518e ＊ , hsa-miR-145 ＊ and hsa-miR-363 ＊ ） ,and 9 down-regulated （hsa-miR-126, hsa-miR-140-3p, hsa-miR-223, hsa-miR- 300, hsa-miR-24-2＊ , hsa-miR-627, hsa-miR-142-3p, h1 sa-miR-744 and hsa-miR-142-5p）. The result of mieroRNA arrays is consistent with the RT-PCR. Conclusion Genes were differentially expressed between BUC and normal bladder mucosa, which may participate in the pathogenesis and development of BUC.
出处
《医学研究生学报》
CAS
2010年第1期48-52,共5页
Journal of Medical Postgraduates
基金
国家自然科学基金(30772278)
南京军区南京总医院青年基金(2008~2010)
