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33
2025年8月21日 星期四
农业生物技术学报  2025, Vol. 33 Issue (1): 157-165    DOI: 10.3969/j.issn.1674-7968.2025.01.014
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
MiR-1306-5p对鸡前脂肪细胞增殖与分化的影响
孙楠桢, 张蕾, 陈若楠, 张若彤, 孙杰*
石河子大学 动物科技学院,石河子 832000
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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摘要 miR-1306-5p对肿瘤细胞的增殖具有促进作用,对脂肪细胞增殖与分化的影响尚不明确。课题组前期采集玫瑰冠鸡(Gallus gallus)和科宝鸡胚胎期第8天胸肌组织进行高通量测序,本研究选择测序数据中筛选出的差异表达的miR-1306-5p,旨在探究miR-1306-5p在鸡前脂肪细胞增殖与分化过程中的功能。利用Targetscan和miRanda软件预测miR-1306-5p的靶基因,以脂肪代谢相关基因—骨形态发生蛋白2 (bone morphogenetic protein 2, BMP2)为靶基因构建野生型和突变型载体,与miR-1306-5p模拟物共转染鸡成纤维细胞,利用荧光素酶报告基因检测系统验证二者的靶标关系。以miR-1306-5p模拟物和抑制物分别转染鸡前脂肪细胞,利用CCK-8 (Cell Counting Kit-8)试剂盒检测细胞增殖能力,通过油红O染色检测油酸诱导的分化情况。结果显示,BMP2野生型与miR-1306-5p模拟物共转染组的荧光素酶活性较阴性对照极显著下降(P<0.01),而突变型没有显著变化;miR-1306-5p模拟物转染鸡前脂肪细胞48、72和96 h,细胞增殖能力极显著低于阴性对照(P<0.01);在鸡前脂肪细胞的油酸诱导分化过程中,脂肪酸结合蛋白 4 (fatty acid binding protein 4, FABP4)基因表达量极显著升高(P<0.01),而内源性miR-1306-5p和BMP2在诱导分化48和72 h的基因表达量极显著降低(P<0.01);在鸡前脂肪细胞中转染miR-1306-5p模拟物后脂滴较对照小,转染miR-1306-5p抑制物后脂滴增多。因此,miR-1306-5p可能通过靶向BMP2而抑制鸡前脂肪细胞的分化。本研究为深入探讨miR-1306-5p在鸡前脂肪细胞分化中的作用机制提供基础资料。
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孙楠桢
张蕾
陈若楠
张若彤
孙杰
关键词 鸡前脂肪细胞miR-1306-5p骨形态发生蛋白2 (BMP2)细胞增殖细胞分化    
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.
Key wordsChicken preadipocytes    miR-1306-5p    Bone morphogenetic protein 2 (BMP2)    Cell proliferation    Cell differentiation
收稿日期: 2024-02-06     
中图分类号: S831.2
基金资助:国家自然科学基金(31660654)
通讯作者: * sunjie_shzu@126.com   
引用本文:   
孙楠桢, 张蕾, 陈若楠, 张若彤, 孙杰. MiR-1306-5p对鸡前脂肪细胞增殖与分化的影响[J]. 农业生物技术学报, 2025, 33(1): 157-165.
SUN Nan-Zhen, ZHANG Lei, CHEN Ruo-Nan, ZHANG Ruo-Tong, SUN Jie. Effect of MiR-1306-5p on the Proliferation and Differentiation of Chicken (Gallus gallus) Preadipocytes. 农业生物技术学报, 2025, 33(1): 157-165.
链接本文:  
https://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2025.01.014     或     https://journal05.magtech.org.cn/Jwk_ny/CN/Y2025/V33/I1/157
 
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