分子营养 Molecular Nutrition

bta-microRNA-145对胰岛素样生长因子1受体-磷脂酰肌醇3激酶-蛋白激酶B/哺乳动物雷帕霉素靶蛋白信号通路相关基因表达的影响及其潜在靶标的揭示

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  • 1. 中国农业科学院北京畜牧兽医研究所, 动物营养学国家重点实验室, 北京 100193;
    2. 河南农业大学生命科学学院, 郑州 450002;
    3. CAAS-ICRAF农用林业与 可持续畜牧业联合实验室, 北京 100193;
    4. 东北农业大学, 食品安全与营养协同创新中心, 哈尔滨 150030
李文清(1979-),女,河南潢川人,博士研究生,研究方向为反刍动物营养。E-mail:liwenqing605@163.com

收稿日期: 2014-04-21

  网络出版日期: 2014-09-10

基金资助

十二五科技支撑计划课题(2012BAD12B02-5);中国农业科学院科技创新工程(ASTIP-IAS07);动物营养学国家重点实验室自主课题(2004DA125184G1103)

Effects of Bta-MicroRNA-145 on IGF1R-PI3K-Akt/mTOR Signal Pathway Related Gene Expressions and Reveal of Its Potential Target

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  • 1. State Key Laboratory of Animal Nutrition, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing 100193, China;
    2. College of Life Science, Henan Agricultural University, Zhengzhou 450002, China;
    3. CAAS-ICRAF Joint Laboratory on Agroforestry and Sustainable Animal Husbandr, Beijing 100193, China;
    4. Synergetic Innovation Center of Safety and Nutrition, Northeast Agricultural University, Harbin 150030, China

Received date: 2014-04-21

  Online published: 2014-09-10

摘要

本文旨在研究bta-microRNA-145(bta-miR-145)对胰岛素样生长因子1受体-磷脂酰肌醇3激酶-蛋白激酶B/哺乳动物雷帕霉素靶蛋白(IGF1R-PI3K-Akt/mTOR)信号通路相关基因表达的影响,为从microRNA的角度研究奶牛的泌乳调控提供依据。以原代奶牛乳腺上皮细胞为研究对象,分别转染bta-miR-145过表达模拟物(mimic)、bta-miR-145抑制表达模拟物(inhibitor),实时定量PCR检测与IGF1R-PI3K-Akt/mTOR信号通路相关的15个基因表达的变化。结果表明:bta-miR-145过表达和抑制表达没有引起预测靶标胰岛素样生长因子1受体(IGF1R)、胰岛素受体底物1(IRS1)、β-酪蛋白(CSN2)基因的mRNA表达量发生显著变化(P>0.05);bta-miR-145过表达可引起真核生物翻译起始因子4E(EIF4E)基因的mRNA表达量的显著下调(P<0.05),抑制表达引起EIF4E基因的mRNA表达量显著上调(P<0.05),并且通过生物信息学分析发现牛EIF4E的3'非翻译区(3'UTR)上存在bta-miR-145的可能结合位点。以上结果提示,bta-miR-145对IGF1R-PI3K-Akt/mTOR信号通路具有一定的调节作用,EIF4E为bta-miR-145的潜在靶标。

本文引用格式

李文清, 王加启, 南雪梅, 卜登攀 . bta-microRNA-145对胰岛素样生长因子1受体-磷脂酰肌醇3激酶-蛋白激酶B/哺乳动物雷帕霉素靶蛋白信号通路相关基因表达的影响及其潜在靶标的揭示[J]. 动物营养学报, 2014 , 26(9) : 2736 -2744 . DOI: 10.3969/j.issn.1006-267x.2014.09.036

Abstract

The object of this experiment was to study the effects of bta-microRNA-145 (bta-miR-145) on the expressions of genes related to insulin-like growth factor 1 receptor-phosphatidylinositol 3-kinase-protein kinase B-mammalian target of rapamycin (IGF1R-PI3K-Akt/mTOR) signaling pathway, and to provide the basis for the regulation of lactation in dairy cow from the perspective of the microRNA. Primary bovine mammary epithelial cells were selected as research object. bta-miR-145 mimic and inhibitor were transfected into cells. After transfection, the expression changes of 15 genes related to IGF1R-PI3K-Akt/mTOR signaling pathway were detected by real-time qPCR. The results showed that the over-expression or down-expression of bta-miR-145 did not cause the expression change of predicted target IGF1R, insulin receptor substrate 1 (IRS1) and beta-casein (CSN2) (P>0.05); the over-expression of bta-miR-145 significantly caused down-regulation of eukaryotic translation initiation factor 4E (EIF4E) mRNA expression (P<0.05), and the down-expression of bta-miR-145 significantly caused increase of EIF4E mRNA expression (P<0.05), and bioinformatics analysis showed that the possible binding site of bovine EIF4E 3'-untranslated region (3'UTR) for bta-miR-145 existed. It is concluded that bta-miR-145 has a role in regulation of IGF1R-PI3K-Akt/mTOR signaling pathway, and EIF4E is a potential target of bta-miR-145.

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