Identification of novel microRNA targets based on microRNA signatures in bladder cancer
Identification of novel microRNA targets based on microRNA signatures in bladder cancer
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DOI：
10.1002/ijc.24390
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发表时间：
2009-07-15
影响因子：
6.4
通讯作者：
Seki, Naohiko
中科院分区：
文献类型：
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作者：
Ichimi, Takahiro;Enokida, Hideki;Seki, Naohiko
MicroRNAs(MiRNAs)是一种对蛋白质编码基因进行负调控的非编码小RNA。为了确定在膀胱癌(BC)中具有肿瘤抑制功能的miRNAs，对14例BCS、5例正常膀胱上皮(NBE)和3个BC细胞系中156个miRNAs进行了筛选。我们发现了7个miRNAs的子集(miR-145、miR-30a-3p、miR-133a、miR-133b、miR-195、miR-125b和miR-1990)在BCS中显著下调。为了证实这些结果，104个BCS和31个NBE进行了基于实时RTPCR的实验，每个miRNA的表达水平在BCS中显著下调(p<；总共0.0001)。受体工作特征曲线分析显示，这些miRNAs的表达水平对区分BC和NBE具有良好的敏感性(>；70%)和特异性(>；75%)。我们的靶点搜索算法和BCS中的基因表达谱(Kawakami等人，Onol Rep 2006；16：521-31)显示，角蛋白7(KRT7mRNA)是下调的miRNAs的共同靶标，并且KRT7mRNA在BCS中的表达水平显著高于NBE(p=0.0004)。Spearman等级相关分析显示KRT7mRNA的表达与每个下调的miRNA之间存在显著的负相关(P<；ALL为0.0001)。功能获得分析表明，3种miRNAs(miR-30-3p、miR-133a和miR-199a*)在BC细胞(KK47)中的表达显著降低。此外，3个miRNAs和si-KRT7均能显著抑制KK47细胞的生长，提示miR-30-3p、miR-133a和miR-199a*可能通过转录抑制KRT7的机制发挥肿瘤抑制作用。(C)2009年UICC
MicroRNAs (miRNAs) are small noncoding RNAs that negatively regulate protein-coding genes. To identify miRNAs that have a tumor suppressive function in bladder cancer (BC), 156 miRNAs were screened in 14 BCs, 5 normal bladder epithelium (NBE) samples and 3 BC cell lines. We identified a subset of 7 miRNAs (miR-145, miR-30a-3p, miR-133a, miR-133b, miR-195, miR-125b and miR-1990) that were significantly downregulated in BCs. To confirm these results, 104 BCs and 31 NBEs were subjected to real-time RT-PCR-based experiments, and the expression levels of each miRNA were significantly downregulated in BCs (p < 0.0001 in all). Receiver-operating characteristic curve analysis revealed that the expression levels of these miRNAs had good sensitivity (>70%) and specificity (>75%) to distinguish BC from NBE. Our target search algorithm and gene-expression profiling in BCs (Kawakami et al., Oncol Rep 2006;16:521-31) revealed that Keratin7 (KRT7) mRNA was a common target of the downregulated miRNAs, and the mRNA expression levels of KRT7 were significantly higher in BCs than in NBEs (p = 0.0004). Spearman rank correlation analysis revealed significant inverse correlations between KRT7 mRNA expression and each downregulated miRNA (p < 0.0001 in all). Gain-of-function analysis revealed that KRT7 mRNA was significantly reduced by transfection of 3 miRNAs (miR-30-3p, miR-133a and miR-199a*) in the BC cell line (KK47). In addition, significant decreases in cell growth were observed after transfection of 3 miRNAs and si-KRT7 in KK47, suggesting that miR-30-3p, miR-133a and miR-199a* may have a tumor suppressive function through the mechanism underlying transcriptional repression of KRT7. (C) 2009 UICC