miR-200b对奶牛乳腺上皮细胞泌乳功能的影响

摘要/Abstract

摘要： 为探索miR-200b对奶牛乳腺上皮细胞泌乳能力的影响，以奶牛乳腺上皮细胞为体外研究模型，利用miR-200b mimics转染进行miR-200b过表达；应用生物信息学软件分析预测miR-200b靶基因，qRT-PCR检测miR-200b过表达后预测靶基因及乳成分合成相关信号通路关键基因mRNA表达的变化；通过CASY细胞活力分析仪及5-乙炔基-2′脱氧尿嘧啶核苷（EdU）标记法分别检测miR-200b过表达对细胞活力及增殖的影响；应用CSN2（β-酪蛋白）试剂盒、TG试剂盒及乳糖试剂盒检测细胞胞外分泌β-酪蛋白、甘油三酯及乳糖含量的变化。生物信息学分析显示，miR-200b与乳腺泌乳功能基因Pten、Dnmt3a、Dnmt3b的3′UTR区序列具有互补结合的位点；qRT-PCR分析结果显示，与对照组相比，过表达miR-200b后的奶牛乳腺上皮细胞中Pten mRNA的表达量显著降低，而乳成分合成相关信号通路关键基因Akt、Srebp1、Csn2、Glut1 mRNA的表达量显著上调；miR-200b 过表达能够提高奶牛乳腺上皮细胞活力，促进奶牛乳腺上皮细胞增殖及β-酪蛋白、甘油三酯和乳糖的合成分泌。综上，miR-200b能够负调控靶基因Pten的表达进而在奶牛乳腺泌乳过程中发挥重要的正调控作用。

关键词: miR-200b, 基因过表达, 奶牛乳腺上皮细胞, Pten, 泌乳调控

边艳杰，韩金汾，段江燕. miR-200b对奶牛乳腺上皮细胞泌乳功能的影响[J]. 河南农业科学, 2018, 47(7): 124-131.

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