| Orginal Article |
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| The Effect of miR-186-5p on the Proliferation and Differentiation of 3T3-L1 Preadipocyte |
GU Hao1,2,GUO Xin-yu1,2,DU Jing-jing1,2,ZHANG Pei-wen1,2,WANG Ding-guo3,LIAO Kun4,ZHANG Shun-hua1,2,ZHU Li1,2,**( ) |
1 Sichuan Agricultural University College of Animal Science and Technology, Chengdu 611130,China
2 Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province,Sichuan Agricultural University, Chengdu 611130,China
3 Chengdu animal disease prevention and control center, Chengdu 610041,China
4 Agricultural Bureau Animal Husbandry Station of Tongjiang, Bazhong 636600,China |
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Abstract Objective: To investigate the effect of miR-186-5p on the proliferation and differentiation of mouse 3t3-L1 preadipocytes and its potential molecular mechanism.Methods: qRT-PCR was utilized to measure the expression levels of miR-186-5p in adipose tissue of different-age mice or during the proliferation and differentiation of 3T3-L1 preadipocyte; miR-186-5p mimics or inhibitors were transfected into the 3T3-L1 adipocytes cultured in growth or differentiation medium using lipofectamine; CCK-8,EdU, qRT-PCR or Oil red O staining were performed to determine the effect of miR-186-5p on 3T3-L1 preadipocyte proliferation or differentiation, respectively. Meanwhile, TargetScan and dual-luciferase assay were used to predict and confirm the target genes of miR-186-5p.Results: With aging of mice or 3T3-L1 preadipocyts differentiation into mature adipocyte, the mRNA levels of miR-186-5p were upregulated gradually. When compared to the negative control, notably, mimics or inhibitors transfection could remarkably increase or decrease the expression levels of miR-186-5p, respectively. MiR-186-5p overexpression significantly inhibited the 3T3-L1 preadipocyte proliferation and promoted its differentiation, and in contrast, miR-186-5p inhibition had opposite effect on 3T3-L1 preadipocyte proliferation and differentiation compared to the overexpression group. Further,overexpression of miR-186-5p significantly repressed the relative luciferase activity of wild type-Wnt5a and Mapk1 3'-UTR were found, whereas its inhibition effect was abolished by the mutation of the binding sites.Conclusions: miR-186-5p inhibits the proliferation of 3T3-L1 preadipocyte and promotes its differentiation by targeting Wnt5a and Mapk1.
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Received: 17 July 2019
Published: 18 April 2020
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Corresponding Authors:
Li ZHU
E-mail: zhuli7508@163.com
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