miR-25-3p, Positively Regulated by Transcription Factor AP-2α, Regulates the Metabolism of C2C12 Cells by Targeting Akt1

• Published in: International journal of molecular sciences Vol. 19; no. 3; p. 773
• Main Authors: Zhang, Feng, Chen, Kun, Tao, Hu, Kang, Tingting, Xiong, Qi, Zeng, Qianhui, Liu, Yang, Jiang, Siwen, Chen, Mingxin
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 08.03.2018; MDPI
• Subjects: 1-Phosphatidylinositol 3-kinase; Adenosine triphosphate; AKT protein; Akt1; AKT1 protein; AP-2α; Assaying; ATP; Binding; Bioinformatics; Biological activity; cell metabolism; Chromatin; Gene expression; Gene regulation; Immunoprecipitation; Kinases; Metabolism; miR-25-3p; mouse; promoter; Protein-serine/threonine kinase; Proteins; Site-directed mutagenesis; Transcription factors; Western blotting; Akt1; miR-25-3p; mouse; cell metabolism; promoter; AP-2α
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms19030773

miR-25, a member of the miR-106b-25 cluster, has been reported as playing an important role in many biological processes by numerous studies, while the role of miR-25 in metabolism and its transcriptional regulation mechanism remain unclear. In this study, gain-of-function and loss-of-function assays demonstrated that miR-25-3p positively regulated the metabolism of C2C12 cells by attenuating phosphoinositide 3-kinase ( ) gene expression and triglyceride (TG) content, and enhancing the content of adenosine triphosphate (ATP) and reactive oxygen species (ROS). Furthermore, the results from bioinformatics analysis, dual luciferase assay, site-directed mutagenesis, qRT-PCR, and Western blotting demonstrated that miR-25-3p directly targeted the AKT serine/threonine kinase 1 ( ) 3' untranslated region (3'UTR). The core promoter of miR-25-3p was identified, and the transcription factor activator protein-2α (AP-2α) significantly increased the expression of mature miR-25-3p by binding to its core promoter in vivo, as indicated by the chromatin immunoprecipitation (ChIP) assay, and AP-2α binding also downregulated the expression of . Taken together, our findings suggest that miR-25-3p, positively regulated by the transcription factor AP-2α, enhances C2C12 cell metabolism by targeting the gene.