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| Effect of MiR-1306-5p on the Proliferation and Differentiation of Chicken (Gallus gallus) Preadipocytes |
| SUN Nan-Zhen, ZHANG Lei, CHEN Ruo-Nan, ZHANG Ruo-Tong, SUN Jie* |
| College of Animal Science and Technology, Shihezi University, Shihezi 832000, China |
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Abstract The functional studies of miR-1306-5p are mostly found to affect the proliferation of smooth muscle cells and various tumor cells. In the early stage, the research group collected the pectoral muscles of rose-crowned chickens (Gallus gallus) and Kebao chickens on the 8th day of embryonic stage for high-throughput sequencing. In this study, differentially expressed miR-1306-5p was selected from the previous sequencing data. The aim was to investigate the function of miR-1306-5p in the proliferation and differentiation of chicken preadipocytes. Using Targetscan and miRanda software predicted target genes of miR-1306-5p. Select the target gene related to fat, bone morphogenetic protein 2 (BMP2), and design the wild type and mutant vector of the target gene BMP2. The vector and miR-1306-5p mimics were constructed to co-transfect chicken fibroblasts, and the target relationship between them was verified by luciferase reporter gene detection system. miR-1306-5p mimics and inhibitors were transfected into chicken preadipocytes, respectively. Cell proliferation capacity was detected by Cell Counting Kit-8 (CCK-8) kits. The differentiation induced by oleic acid was detected by oil red O staining. The results showed that the luciferase activity in the BMP2 wild type co-transfected with miR-1306-5p mimics group was extremely significantly decreased compared with the negative control group (P<0.01), while the mutant type had no significant change. After transfection with miR-1306-5p mimics for 48, 72 and 96 h, the proliferation capacity of chicken preadipocytes was extremely significantly lower than that of the negative control group (P<0.01). The gene expression of fatty acid binding protein 4 (FABP4) was extremely significantly increased during oleic acid-induced differentiation of chicken preadipocytes (P<0.01). The gene expressions of endogenous miR-1306-5p and BMP2 were extremely significantly decreased at 48 and 72 h after induction of differentiation (P<0.01). Lipid droplets were smaller after transfection of miR-1306-5p mimics and increased after transfection of miR-1306-5p inhibitor in chicken preadipocytes. Therefore, miR-1306-5p may inhibit the proliferation and differentiation of chicken preadipocytes by targeting BMP2. This study provides basic data for further exploring the mechanism of miR-1306-5p in chicken preadipocyte differentiation.
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Received: 06 February 2024
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Corresponding Authors:
* sunjie_shzu@126.com
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