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

Abstract

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 (PI3K) 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 (Akt1) 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 Akt1. Taken together, our findings suggest that miR-25-3p, positively regulated by the transcription factor AP-2α, enhances C2C12 cell metabolism by targeting the Akt1 gene.

RETRACTED: Platelet-Derived Exosomal MicroRNA-25-3p Inhibits Coronary Vascular Endothelial Cell Inflammation Through Adam10 via the NF-κB Signaling Pathway in ApoE−/− Mice

“…MiR-25-3p can be defined as the exudation of liposarcoma cells via extracellular vesicles, which interacts with the surrounding microenvironment to enhance cell metastasis, invasion, and proliferation in liposarcoma (13). Besides, miR-25-3p has recently been revealed to modulate C2C12 cell metabolism through its regulation of AKT1 (14). In addition, it has been noted that many miR-25 target mRNAs are implicated in a variety of biological procedures, including inflammation (15).…”

The biological characteristics of transcription factors AP-2α and AP-2γ and their importance in various types of cancers

“…AP-2α was listed as a regulator of at least two miRNAs that are responsible for either metabolism regulation or proper craniofacial phenotype (or disrupted when gene is mutated). In the first case, AP-2α binds to the core promoter of miR-25-3p, thus increasing its expression and allowing it to target the Akt1 gene and enhance the metabolism in C2C12 cells [ 80 ]. In the second, it recognizes multiple binding sites in miR-17-92 chromatin that induce miRNA expression and allow proper midface development through regulation of Tbx genes.…”

The roles of microRNAs related with progression and metastasis in human cancers

“…组织中显著降低, 其可通过抑制AKT1的表达从而阻 遏神经胶质瘤细胞的生长和侵袭 [30] . 此外, miR-25-3P [31] 和miR-302-367簇 [32] 可通过调控AKT1的表达, 进…”