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Exploration of targets regulated by miR-125b in porcine adipocytes

Author:
Cheng, Xiao, Chen, Xingping, Wang, Peng, Chen, Ting, Sun, Jiajie, Xi, Qianyun, Zhang, Yongliang
Source:
In vitro cellular & developmental biology 2020 v.56 no.2 pp. 103-111
ISSN:
1071-2690
Subject:
acyl-CoA dehydrogenase, adipocytes, adipose tissue, algorithms, enzyme activity, enzyme inhibition, fatty acid metabolism, fatty acids, gene expression, gene expression regulation, homeostasis, luciferase, messenger RNA, mice, microRNA, mitochondria, prediction, protein synthesis, quantitative polymerase chain reaction, swine
Abstract:
MicroRNA (miRNA) has been proved to play a key role in lipid metabolism. In our previous study, miR-125b was validated to be differentially expressed in preadipocytes and adipocytes, which was also proved to involve in lipid metabolism. To explore the comprehensive targets of miR-125b in adipocytes, isobaric tag for relative and absolute quantitation (iTRAQ) analysis was performed to obtain differentially expressed proteins in adipocytes comparing negative control (NC) and miR-125b mimic, combining with digital gene expression (DGE) profiling of mRNA incorporated into RNA-induced silencing complex (RISC) pulled down by biotinylated miR-125b mimic and targets prediction of miR-125b by three algorithms, acyl-CoA dehydrogenase short chain (ACADS) and mitochondrial trans-2-enoyl-CoA reductase (MECR) were screened out as miR-125b direct targets. Luciferase reporter assay further validated that miR-125b mimic significantly inhibited the luciferase activity by targeting wild type (WT) 3′-UTR compared with NC. qPCR analysis of ACADS and MECR mRNA from adipose tissues of miR-125b knockout (KO) mice further confirmed the inhibition of miR-125b on ACADS and MECR expressions. Here we report miR-125b play a vital role in maintaining homeostasis of fatty acid metabolism by targeting key enzyme ACADS and MECR in the process of fatty acid elongation and degradation.
Agid:
6856804