中国畜牧兽医

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microRNA-152对奶牛乳腺上皮细胞乳蛋白合成的调控作用

王杰,边艳杰,王卓然,李丹,高学军,李庆章,王春梅   

  1. (东北农业大学乳品科学教育部重点实验室,黑龙江哈尔滨  150030

  • 修回日期:2014-02-27 出版日期:2014-06-20 发布日期:2014-07-24
  • 通讯作者: 王春梅,女,博士,教授,硕士生导师,从事乳业生物技术领域的研究。E-mail:wangcm-1@163.com
  • 作者简介:王杰(1985—),男,河北人,硕士生,研究方向:泌乳生物学与乳腺功能调控。
  • 基金资助:

    国家重点基础研究发展规划(973)项目(2011CB100804)。

Effects of microRNA-152 on Milk Protein Synthesis in Bovine Mammary Epithelial Cells

WANG Jie, BIAN Yan-jie, WANG Zhuo-ran, LI Dan, GAO Xue-jun, LI Qing-zhang, WANG Chun-mei   

  1.  (Key Laboratory of Dairy Sciences, Ministry of Education, Northeast Agricultural University, Harbin 150030, China)

  • Revised:2014-02-27 Online:2014-06-20 Published:2014-07-24

摘要: 为确定microRNA-152(miR-152)对奶牛乳腺上皮细胞中乳蛋白合成的影响,本试验应用脂质体转染技术,改变miR-152在奶牛乳腺上皮细胞的表达量。采用实时荧光定量PCR、Western blotting、细胞活力分析等技术探索miR-152对奶牛乳腺上皮细胞增殖及泌乳功能的影响。结果显示,miR-152沉默,细胞因子信号传导抑制蛋白3(suppressor of cytokine signaling 3,SOCS3)表达增强,信号转导通路分子磷酸化信号转导与转录激活因子5(phospho-signal transducer and activator of transcription 5,p-STAT5)、磷酸化哺乳动物雷帕霉素蛋白(phospho-mammalian target of rapamycin,p-mTOR)、核糖体S6激酶1(ribosomal protein S6 kinase 1,S6K1)、细胞周期蛋白D1(cyclinD1)表达减弱,细胞增殖能力减弱,β-酪蛋白分泌减少。研究结果表明,miR-152在奶牛乳腺上皮细胞调控的靶基因是SOCS3,通过抑制其表达而发挥作用。miR-152可通过调控p-STAT5、p-mTOR、S6K1、cyclinD1信号转导而促进乳蛋白的合成和乳腺上皮细胞增殖。

关键词:

奶牛乳腺上皮细胞; 增殖; 泌乳; miRmso-bidi-font-family: 宋体" lang="EN-US">-152; 细胞因子信号传导抑制蛋白3

Abstract: In order to study the effect of microRNA-152 (miR-152) on milk protein synthesis in dairy cow mammary epithelial cells, liposome transfection was used to change the expression of miR-152 in bovine mammary epithelial cells. The influence of miR-152 on the cell proliferation and lactation was detected using Real-time PCR, Western blotting, cell viability analysis and other techniques. Results indicated that when miR-152 was inhibited, SOCS3 protein expression was increased, signaling pathway factors p-STAT5,p-mTOR,S6K1 and cyclinD1 protein expression were decreased, cell proliferation was decreased, β-casein secretion was decreased. The results suggested that miR-152 probably regulated bovine mammary epithelial cells by suppressing expression of SOCS3. And miR-152 could increase milk protein synthesis and cells proliferation by regulating signal transductions of p-STAT5,p-mTOR,S6K1 and cyclinD1 in mammary epithelial cells.

Key words:

bovine mammary epithelial cellsproliferationlactationmiRmso-bidi-font-family: 宋体" lang="EN-US">-152suppressor of cytokine signaling 3 (SOCS3)