Department of Molecular Genetics, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama, Japan.
Proteomics. 2011 Sep;11(17):3531-9. doi: 10.1002/pmic.201000501. Epub 2011 Aug 1.
MicroRNAs (miRNAs) are small non-coding RNAs that post-transcriptionally repress the expression of target genes. Many miRNAs have been implicated in a number of diseases, including cancers. The miR-17-92 miRNA cluster is known as a body of oncogenic miRNAs, and has been shown to be overexpressed in several cancers, including lung cancer. Although the overexpression of miR-17-92 is clearly implicated in the development of lung cancer, only a few direct targets for the miR-17-92 cluster have been identified thus far. In this study, we examined miR-17-92 target profiles in SBC-3 small-cell lung cancer cells using a quantitative proteomic strategy to identify direct targets of the miR-17-92 cluster. By knocking down the expression of endogenous miR-19a, miR-20a and miR-92-1, which are contained in the cluster, 112 up-regulated proteins were detected and also identified as potential targets of these miRNAs. Among these candidate targets, we validated one direct target, RAB14. In conclusion, these findings suggest that proteomic approaches are valuable for identifying direct miRNA targets, and we were able to identify a novel direct target for the miR-92-1 using our proteomic strategy.
微小 RNA(miRNA)是一种小型非编码 RNA,可在后转录水平上抑制靶基因的表达。许多 miRNA 与许多疾病有关,包括癌症。miR-17-92 miRNA 簇被称为致癌 miRNA 群,已被证明在多种癌症中过度表达,包括肺癌。尽管 miR-17-92 的过表达显然与肺癌的发展有关,但迄今为止仅鉴定出 miR-17-92 簇的少数几个直接靶标。在这项研究中,我们使用定量蛋白质组学策略检查了 SBC-3 小细胞肺癌细胞中 miR-17-92 的靶标谱,以鉴定 miR-17-92 簇的直接靶标。通过敲低包含在簇中的内源性 miR-19a、miR-20a 和 miR-92-1 的表达,检测到 112 个上调蛋白,并将其鉴定为这些 miRNA 的潜在靶标。在这些候选靶标中,我们验证了一个直接靶标 RAB14。总之,这些发现表明蛋白质组学方法对于鉴定直接 miRNA 靶标非常有价值,并且我们能够使用我们的蛋白质组学策略鉴定 miR-92-1 的一个新的直接靶标。