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

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 assay...

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Autores principales: Zhang, Feng, Chen, Kun, Tao, Hu, Kang, Tingting, Xiong, Qi, Zeng, Qianhui, Liu, Yang, Jiang, Siwen, Chen, Mingxin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5877634/
https://www.ncbi.nlm.nih.gov/pubmed/29518009
http://dx.doi.org/10.3390/ijms19030773
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author Zhang, Feng
Chen, Kun
Tao, Hu
Kang, Tingting
Xiong, Qi
Zeng, Qianhui
Liu, Yang
Jiang, Siwen
Chen, Mingxin
author_facet Zhang, Feng
Chen, Kun
Tao, Hu
Kang, Tingting
Xiong, Qi
Zeng, Qianhui
Liu, Yang
Jiang, Siwen
Chen, Mingxin
author_sort Zhang, Feng
collection PubMed
description 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.
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spelling pubmed-58776342018-04-09 miR-25-3p, Positively Regulated by Transcription Factor AP-2α, Regulates the Metabolism of C2C12 Cells by Targeting Akt1 Zhang, Feng Chen, Kun Tao, Hu Kang, Tingting Xiong, Qi Zeng, Qianhui Liu, Yang Jiang, Siwen Chen, Mingxin Int J Mol Sci Article 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. MDPI 2018-03-08 /pmc/articles/PMC5877634/ /pubmed/29518009 http://dx.doi.org/10.3390/ijms19030773 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Zhang, Feng
Chen, Kun
Tao, Hu
Kang, Tingting
Xiong, Qi
Zeng, Qianhui
Liu, Yang
Jiang, Siwen
Chen, Mingxin
miR-25-3p, Positively Regulated by Transcription Factor AP-2α, Regulates the Metabolism of C2C12 Cells by Targeting Akt1
title miR-25-3p, Positively Regulated by Transcription Factor AP-2α, Regulates the Metabolism of C2C12 Cells by Targeting Akt1
title_full miR-25-3p, Positively Regulated by Transcription Factor AP-2α, Regulates the Metabolism of C2C12 Cells by Targeting Akt1
title_fullStr miR-25-3p, Positively Regulated by Transcription Factor AP-2α, Regulates the Metabolism of C2C12 Cells by Targeting Akt1
title_full_unstemmed miR-25-3p, Positively Regulated by Transcription Factor AP-2α, Regulates the Metabolism of C2C12 Cells by Targeting Akt1
title_short miR-25-3p, Positively Regulated by Transcription Factor AP-2α, Regulates the Metabolism of C2C12 Cells by Targeting Akt1
title_sort mir-25-3p, positively regulated by transcription factor ap-2α, regulates the metabolism of c2c12 cells by targeting akt1
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5877634/
https://www.ncbi.nlm.nih.gov/pubmed/29518009
http://dx.doi.org/10.3390/ijms19030773
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