Department of Systems BioMedicine, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University (TMDU), Bunkyo, Tokyo 113-8519, Japan.
Department of Systems BioMedicine, National Research Institute for Child Health and Development, Setagaya, Tokyo 157-8535, Japan.
Proc Natl Acad Sci U S A. 2017 Apr 11;114(15):3927-3932. doi: 10.1073/pnas.1620019114. Epub 2017 Mar 29.
miRNAs play critical roles in various biological processes by targeting specific mRNAs. Current approaches to identifying miRNA targets are insufficient for elucidation of a miRNA regulatory network. Here, we created a cell-based screening system using a luciferase reporter library composed of 4,891 full-length cDNAs, each of which was integrated into the 3' UTR of a luciferase gene. Using this reporter library system, we conducted a screening for targets of miR-34a, a tumor-suppressor miRNA. We identified both previously characterized and previously uncharacterized targets. miR-34a overexpression in MDA-MB-231 breast cancer cells repressed the expression of these previously unrecognized targets. Among these targets, is crucial for MDA-MB-231 cell growth, and its expression correlated with the overall survival of patients with breast cancer. Furthermore, was found to be directly regulated by miR-34a via its coding region. These data show that this system is useful for elucidating miRNA functions and networks.
miRNAs 通过靶向特定的 mRNAs 在各种生物过程中发挥关键作用。目前鉴定 miRNA 靶标的方法不足以阐明 miRNA 调控网络。在这里，我们创建了一个基于细胞的筛选系统，该系统使用由 4891 个全长 cDNA 组成的荧光素酶报告基因文库，每个 cDNA 都整合到荧光素酶基因的 3'UTR 中。使用这个报告基因文库系统，我们对 miR-34a（一种肿瘤抑制 miRNA）的靶标进行了筛选。我们鉴定了以前表征和以前未表征的靶标。在 MDA-MB-231 乳腺癌细胞中过表达 miR-34a 会抑制这些以前未识别的靶标的表达。在这些靶标中， 对于 MDA-MB-231 细胞的生长至关重要，其表达与乳腺癌患者的总生存率相关。此外，发现 通过其编码区直接受 miR-34a 调控。这些数据表明该系统可用于阐明 miRNA 的功能和网络。
