MicroRNA-148a is down-regulated in human pancreatic ductal adenocarcinomas and regulates cell survival by targeting CDC25B
MicroRNA-148a is down-regulated in human pancreatic ductal adenocarcinomas and regulates cell survival by targeting CDC25B
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DOI：
10.1038/labinvest.2011.99
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发表时间：
2011-10-01
影响因子：
5
通讯作者：
Tannapfel, Andrea
中科院分区：
文献类型：
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作者：
Liffers, Sven-T;Munding, Johanna B.;Tannapfel, Andrea
MicroRNA（miRNAs：短的非编码RNA）正在成为一类潜在的新型肿瘤标志物，因为在各种类型的癌症中越来越多地报道了它们的失调。在本研究中，我们研究了miRNA-148 a（miR-148 a）在人胰腺导管腺癌（PDAC）中的转录状态及其在双特异性蛋白磷酸酶CDC 25 B调节中的作用。我们观察到miR-148 a在PDAC中表现出与正常胰腺导管细胞相反的4倍显著下调。此外，我们观察到慢病毒介导的miR-148 a在胰腺癌细胞系IMIM-PC 2中的稳定过表达抑制了肿瘤细胞的生长和集落形成。此外，通过使用PicTar、Targetscan和米兰达结合基因本体分析的计算机分析，将CDC 25 B鉴定为miR-148 a的潜在靶标。通过体外荧光素酶测定验证了miR-148 a与CDC 25 B的30个非翻译区（UTR）之间的拟议相互作用。我们证明了在miR-148 a模拟物存在下，含有CDC 25 B的3 'UTR的荧光素酶报告基因的活性被抑制，证实了miR-148 a靶向CDC 25 B的3' UTR。最后，在过表达miR-148 a的IMIM-PC 2-细胞、瞬时转染的胰腺细胞系（通过Western印迹分析检测）以及患者肿瘤样品（通过免疫组织化学检测）中，CDC 25 B在蛋白质水平下调。总之，我们确定了CDC 25 B作为一种新的miR-148 a靶点，它可能在PDAC中具有增殖优势。实验室调查（2011）91，1472-1479; doi：10.1038/labinvest.2011.99;在线发表2011年6月27日
MicroRNAs (miRNAs: short non-coding RNAs) are emerging as a class of potential novel tumor markers, as their dysregulation is being increasingly reported in various types of cancers. In the present study, we investigated the transcription status of miRNA-148a (miR-148a) in human pancreatic ductal adenocarcinoma (PDAC) and its role in the regulation of the dual specificity protein phosphatase CDC25B. We observed that miR-148a exhibited a significant 4-fold down-regulation in PDAC as opposed to normal pancreatic ductal cells. In addition, we observed that stable lentiviral-mediated overexpression of miR-148a in the pancreatic cancer cell line IMIM-PC2, inhibited tumor cell growth and colony formation. Furthermore, CDC25B was identified as a potential target of miR-148a by in silico analysis using PicTar, Targetscan and miRanda in conjunction with gene ontology analysis. The proposed interaction between miR-148a and the 30 untranslated region (UTR) of CDC25B was verified by in-vitro luciferase assays. We demonstrate that the activity of a luciferase reporter containing the 3'UTR of CDC25B was repressed in the presence of miR-148a mimics, confirming that miR-148a targets the 3'UTR of CDC25B. Finally, CDC25B was down-regulated at the protein level in miR-148a overexpressing IMIM-PC2-cells, and in transiently transfected pancreatic cell lines (as detected by Western blot analysis), as well as in patient tumor samples (as detected by immunohistochemistry). In summary, we identified CDC25B as a novel miR-148a target which may confer a proliferative advantage in PDAC. Laboratory Investigation (2011) 91, 1472-1479; doi:10.1038/labinvest.2011.99; published online 27 June 2011