《中国畜牧兽医》 ›› 2017, Vol. 44 ›› Issue (5): 1267-1274.doi: 10.16431/j.cnki.1671-7236.2017.05.002

• 生物技术 • 上一篇    下一篇

METTL3对乳腺上皮细胞中乳蛋白和乳脂肪合成的影响

李冬, 黄鑫, 李姗姗, 于萌萌, 李萌, 张明辉, 高学军   

  1. 东北农业大学乳品科学教育部重点实验室, 哈尔滨 150030
  • 收稿日期:2016-09-20 出版日期:2017-05-20 发布日期:2017-05-18
  • 通讯作者: 高学军 E-mail:gaoxj5390@sina.com
  • 作者简介:李冬(1992-),女,黑龙江鸡西人,硕士生,研究方向:生物化学与分子生物学,E-mail:dqsflidong@126.com
  • 基金资助:

    国家自然科学基金项目(31472162);国家高技术研究发展计划(863计划)项目(2013AA102504-03)

Effect of METTL3 on Milk Protein and Milk Fat Synthesis of Bovine Mammary Epithelial Cells

LI Dong, HUANG Xin, LI Shan-shan, YU Meng-meng, LI Meng, ZHANG Ming-hui, GAO Xue-jun   

  1. Key Laboratory of Dairy Science of Education Ministry, Northeast Agricultural University, Harbin 150030, China
  • Received:2016-09-20 Online:2017-05-20 Published:2017-05-18

摘要:

本试验旨在研究m6A甲基转移酶(methyltransferase-like protein 3,METTL3)在奶牛乳腺上皮细胞(bovine mammary epithelial cells,BMEC)中对乳蛋白和乳脂肪的调节作用。试验建立不同的BMEC模型,应用Western blotting、免疫荧光法检测添加雌激素(E)、蛋氨酸(Met)、METTL3超表达和抑制时对BMEC中乳蛋白和乳脂的影响,以及基因定位和表达变化。结果显示,当在BMEC培养液中添加E、Met及METTL3表达水平提高时,mTOR、p-mTOR、GlyRS、p-GlyRS、CSN2、SREBP-1c等表达量均增加,反之,这些基因表达量均下降,表明METTL3可以通过调控乳脂、乳蛋白的相关信号通路分子的基因表达进而影响乳脂、乳蛋白的表达。提示METTL3是乳合成的重要调节分子,可对细胞中乳脂、乳蛋白的合成起正调控作用。

关键词: m6A甲基转移酶; 奶牛乳腺上皮细胞; 乳脂肪; 乳蛋白

Abstract:

In this study,to investigate the regulatory role of methyltransferase-like protein 3 (METTL3) on milk protein and fat synthesis in bovine mammary epithelial cells (BMEC),different BMEC models were established, the expression of related genes and localization of METTL3 were detected,and the impact of METTL3 on milk protein and fat synthesis in BMEC after adding β-estrogen (E),methionine (Met),and overexpression or inhibition of METTL3 were measured using Western blotting and imunofluorescience methods. The results showed that when E and Met were added or the METTL3 expression was promoted,the expressions of mTOR,p-mTOR,GlyRS,p-GlyRS,CSN2 and SREBP-1c were increased,oppositely,the gene expression were decreased. Those results indicated that METTL3 could affect the milk fat and protein synthesis by adjusting the expression of related signaling pathway gene.METTL3 was one of the important regulating milk synthetic molecules and had an positively regulation on milk fat and protein synthesis in cells.

Key words: m6A methyltransferase-like protein 3(METTL3); bovine mammary epithelial cells(BMEC); milk fat; milk protein

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