膀胱尿路上皮癌差异表达微小RNA的筛选、验证及意义
Detection, verification and significance of differentially expressed miRNAs in bladder urotheliai carcinoma
摘要目的 分析膀胱尿路上皮癌中差异表达的微小RNA(miRNA),为进一步研究相关miRNA的功能及致癌机制提供平台.方法 收集2008年9月至2009年9月南京军区南京总医院泌尿外科手术获得标本,5份正常膀胱黏膜上皮组织和30例膀胱尿路上皮癌患者膀胱组织(其中浸润癌18例、非浸润癌12例;Ⅰ级10例、Ⅱ级10例、Ⅲ级10例),miRNA芯片分析筛选出差异表达miRNA.重新收集正常膀胱黏膜上皮组织(n=10)和膀胱尿路上皮癌标本(n=50);选4型膀胱尿路上皮癌细胞株,用反转录聚合酶链反应(RT-PCR)验证miRNA基因芯片结果.结果 Ⅰ级、Ⅱ级、Ⅲ级、Ⅰ+Ⅱ+Ⅲ级、浸润性、非浸润性膀胱尿路上皮癌分别与正常膀胱黏膜上皮比较所得的6组差异表达基因中,has-miR-29b-1*均上调,has-miR-300、has-miR-923均下调.浸润性与非浸润性膀胱尿路上皮癌相比有7个差异表达基因,上调6个,下调1个.RT-PCR验证结果 与基因芯片结果 一致;4型膀胱癌细胞株中,T24细胞株RT-PCR验证结果 与基因芯片结果 完全一致.结论 膀胱尿路上皮癌与正常膀胱黏膜上皮组织比较存在差异表达的miRNA,其可能与膀胱尿路上皮癌的发生、浸润、进展密切相关;膀胱尿路上皮癌T24细胞株与膀胱尿路上皮癌有一致的miRNA差异表达.
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abstractsObjective To explore the differentially expressed miRNAs in bladder urothelial carcinoma (BUC) in order to provide rationales for further studies of function and mechanism for their actions. Methods Fresh normal(5 samples) and BUC(30 samples) tissues were collected from September 2008 to September 2009 in our hospital and analyzed by miRNA microarray to find differentially expressed miRNAs; RT-PCR (reverse transcription-polymerase chain reaction) of recollected samples and 4 types of cell strain to verify the selected miRNAs. Results In grade Ⅰ , grade Ⅱ , grade Ⅲ, grade Ⅰ + Ⅱ + Ⅲ ,infiltrating and non-infiltrating groups, hsa-miR-29b-1 * was up-regulated while hsa-miR-923 and hsa-miR300 were down-regulated. When the infiltrating BUC group was compared with the non-infiltrating BUC group, there were 7 differentially expressed miRNAs. In collected samples, the result of RT-PCR was consistent with that of miRNA array. In 4 types of cancer cell strains, only the result of T24 was completely consistent with that of miRNA array. Conclusions There are differentially expressed miRNAs between normal bladder mucosa and BUC, infiltration BUC and non-infiltration BUC. The result of T24 cell strain is completely consistent with that of miRNA array.
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