Department of Medicinal Chemistry, School of Pharmacy, Xi'an Jiaotong University Health Science Center, Xi'an, Shaanxi, People's Republic of China.
Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Xi'an Jiaotong University Health Science Center, Xi'an, Shaanxi, People's Republic of China.
Biotechnol Appl Biochem. 2019 Sep;66(5):755-762. doi: 10.1002/bab.1756. Epub 2019 Aug 7.
The therapeutic potential of microRNA-21 (miR-21) small-molecule inhibitors has been of particular interest to medicinal chemists. Moreover, the development of more facile screening methods is lacking. In the present study, two potential screening strategies for miR-21 small-molecule inhibitor including the stem-loop reverse transcription-quantitative PCR and dual luciferase reporter assay system were demonstrated and discussed in detail. A pmirGLO-miR21cswt plasmid and its two different mutants were constructed for dual luciferase reporter assay system. In addition, the sensitivity and specificity of these two methods were validated. Our results demonstrated that both strategies are decent choices for the screening of small-molecule inhibitors for miR-21 and possibly other miRNAs. Eventually, we applied our optimized strategy to discover and characterize several promising compounds such as azobenzene derivate A, enoxacin, and norfloxacin for their potential impact on intracellular miR-21 concentration.
miR-21 小分子抑制剂的治疗潜力引起了药物化学家的特别关注。此外，缺乏更简便的筛选方法。在本研究中，展示并详细讨论了两种用于 miR-21 小分子抑制剂的潜在筛选策略，包括茎环逆转录定量 PCR 和双荧光素酶报告基因检测系统。构建了 pmirGLO-miR21cswt 质粒及其两个不同的突变体用于双荧光素酶报告基因检测系统。此外，验证了这两种方法的灵敏度和特异性。我们的结果表明，这两种策略都是筛选 miR-21 和可能其他 miRNA 的小分子抑制剂的不错选择。最终，我们应用优化后的策略发现并表征了几种有前途的化合物，如偶氮苯衍生物 A、依诺沙星和诺氟沙星，以研究它们对细胞内 miR-21 浓度的潜在影响。
