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

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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.

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Funding

This research was supported by grants from the National Key Research and Development Program of China (2016YFD0500503), and the Natural Science Foundation of China program (31472163).

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Authors and Affiliations

  1. National Engineering Research Center for Breeding Swine Industry, College of Animal Science, South China Agricultural University, Guangzhou, 510642, China

    Xiao Cheng, Xingping Chen, Peng Wang, Ting Chen, Jiajie Sun, Qianyun Xi & Yongliang Zhang

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  1. Xiao Cheng
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  2. Xingping Chen
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  3. Peng Wang
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  4. Ting Chen
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  5. Jiajie Sun
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  6. Qianyun Xi
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  7. Yongliang Zhang
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Corresponding author

Correspondence to Yongliang Zhang.

Ethics declarations

All the animal experiments were conducted in accordance with the guidelines of Guangdong Province on the Review of Welfare and Ethics of Laboratory Animals approved by the Guangdong Province Administration Office of Laboratory Animals (GPAOLO). All animal procedures were conducted under the protocol (SCAU-AEC-2010-0416) approved by the Animal Ethics Committee of South China Agricultural University.

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The authors declare that they have no conflict of interest.

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Editor: Tetsuji Okamoto

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Cheng, X., Chen, X., Wang, P. et al. Exploration of targets regulated by miR-125b in porcine adipocytes. In Vitro Cell.Dev.Biol.-Animal 56, 103–111 (2020). https://doi.org/10.1007/s11626-019-00427-3

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  • DOI: https://doi.org/10.1007/s11626-019-00427-3

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