miR-142和miR-144靶向FoxO1基因调节绵羊前体脂肪细胞分化的研究

doi:10.11843/j.issn.0366-6964.2018.04.003

畜牧兽医学报 ›› 2018, Vol. 49 ›› Issue (4): 675-684.doi: 10.11843/j.issn.0366-6964.2018.04.003

miR-142和miR-144靶向FoxO1基因调节绵羊前体脂肪细胞分化的研究

赵艳艳, 乔利英, 景炅婕, 潘洋洋, 任端阳, 王书芳, 吕宣增, 刘文忠*

山西农业大学动物科技学院, 太谷 030801

收稿日期:2017-09-12 出版日期:2018-04-23 发布日期:2018-04-23
通讯作者: 刘文忠,教授,博士生导师,主要从事动物遗传资源的分子评价与种质创新研究,E-mail:tglwzyc@163.com
作者简介:赵艳艳(1992-),女,山西长治人,硕士生,主要从事肉用绵羊的分子遗传育种研究,E-mail:yanyanzhao163@163.com。
基金资助:
山西优势肉用家畜高效安全生产协同创新中心项目；山西农业大学拔尖创新人才支持计划项目（BJRC201203）

Study on the Differentiation Regulation of miR-142 and miR-144 on Ovine Preadipocytes by Targeting FoxO1 Gene

ZHAO Yan-yan, QIAO Li-ying, JING Jiong-jie, PAN Yang-yang, REN Duan-yang, WANG Shu-fang, LÜ Xuan-zeng, LIU Wen-zhong*

College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu 030801, China

Received:2017-09-12 Online:2018-04-23 Published:2018-04-23
Supported by:

摘要/Abstract

摘要：

旨在预测并验证调节FoxO1基因表达的关键miRNAs及其调节脂肪分化的机制。本研究利用4个在线软件预测与FoxO1基因具有潜在靶标关系的miRNAs。用双荧光素酶报告系统检测荧光活性。用荧光定量PCR、Western blotting和油红O染色，分别在mRNA和蛋白水平上检测miRNA对FoxO1表达的影响以及对绵羊前体脂肪细胞分化的调节作用。结果表明，miR-142和miR-144与FoxO1具有靶标关系。过表达miR-142和miR-144显著抑制了双荧光素酶报告载体的荧光活性，下调了FoxO1 mRNA（P<0.01）及其编码蛋白的表达（P<0.05）。miR-142和miR-144均可促进绵羊前体脂肪细胞分化，加快脂滴形成。由此证明，miR-142和miR-144靶向负调节FoxO1的表达，进而通过FoxO1对PPARγ的负调节促进绵羊前体脂肪细胞的分化。

Abstract:

This study aimed to predict and validate the crucial miRNAs that regulate FoxO1 gene expression, and to elucidate the functional mechanism of these miRNAs in ovine adipose differentiation.Four online softwares were used to predict the miRNAs that might target FoxO1. A dual-luciferase reporter assay system was employed to detect luciferase activity. qPCR, Western blotting and Oil red O staining were used to detect the effect of these miRNAs on the expression of FoxO1 both at mRNA and protein levels, and on the regulation of ovine preadipocyte differentiation. The results showed that FoxO1 was targeted by both miR-142 and miR-144.Overexpression of miR-142 or miR-144 significantly inhibited the luciferase activity of the dual-luciferase reporter vector, and reduced the expression of FoxO1 mRNA (P<0.01) and protein (P<0.05). Both miR-142 and miR-144 promoted the differentiation of ovine preadipocytes and accelerated the formation of lipid droplets. In conclusion, miR-142 and miR-144 can down-regulate the expression of FoxO1, and further down-regulate PPARγ expression,thus promote the differentiation of ovine preadipocytes.

中图分类号:

S826.2

赵艳艳, 乔利英, 景炅婕, 潘洋洋, 任端阳, 王书芳, 吕宣增, 刘文忠. miR-142和miR-144靶向FoxO1基因调节绵羊前体脂肪细胞分化的研究[J]. 畜牧兽医学报, 2018, 49(4): 675-684.

ZHAO Yan-yan, QIAO Li-ying, JING Jiong-jie, PAN Yang-yang, REN Duan-yang, WANG Shu-fang, Lü Xuan-zeng, LIU Wen-zhong. Study on the Differentiation Regulation of miR-142 and miR-144 on Ovine Preadipocytes by Targeting FoxO1 Gene[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(4): 675-684.

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miR-142和miR-144靶向FoxO1基因调节绵羊前体脂肪细胞分化的研究

Study on the Differentiation Regulation of miR-142 and miR-144 on Ovine Preadipocytes by Targeting FoxO1 Gene

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