MiR-16a Regulates Milk Fat Metabolism by Targeting Large Tumor Suppressor Kinase 1 (LATS1) in Bovine Mammary Epithelial Cells
MiR-16a Regulates Milk Fat Metabolism by Targeting Large Tumor Suppressor Kinase 1 (LATS1) in Bovine Mammary Epithelial Cells
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MiR-16a 通过靶向牛乳腺上皮细胞中的大肿瘤抑制激酶 1 (LATS1) 调节乳脂代谢
DOI：
10.1021/acs.jafc.9b04883
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发表时间：
2019-10-09
影响因子：
6.1
通讯作者：
Yang, Zhangping
中科院分区：
文献类型：
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作者：
Chen, Zhi;Chu, Shuangfeng;Yang, Zhangping
牛奶含有许多有益的脂肪酸，包括短链和中链以及不饱和的共轭和非共轭脂肪酸。在这项研究中，microRNA测序的乳腺组织收集在早期，高峰期，中期和后期的泌乳期，以确定miRNA的表达谱。miR-16 a是差异表达的miRNA之一，并被选择用于与脂肪酸代谢有关的深入功能研究。miR-16 a的模拟物损害了脂肪代谢[三酰甘油（TAG）和胆固醇]，而miR-16 a的敲低促进了牛乳腺上皮细胞（BMEC）的体外脂肪代谢。此外，对BMEC的体外研究还显示，miR-16 a对顺式9-C18：1、总C18：1、C20：1和C22：1以及长链多不饱和脂肪酸的细胞浓度具有负面影响。因此，这些表明对脂肪酸代谢的负面影响的数据扩展了miR-16 a在介导脂肪细胞分化中的关键作用的发现。通过生物信息学分析、靶基因3' UTR荧光素酶报告基因分析和蛋白质印迹分析的组合，我们鉴定了大肿瘤抑制激酶1（LATS 1）作为miR-16 a的靶点。将siRNA-LATS 1转染入BMEC导致TAG、胆固醇和细胞脂肪酸浓度的增加，表明LATS 1在乳腺细胞脂肪酸代谢中的积极作用。总之，数据表明miR-16 a通过LATS 1调节与细胞内TAG、胆固醇和不饱和脂肪酸合成相关的生物学过程。这些数据为进一步阐明奶牛乳腺细胞脂质代谢的调控提供了理论和实验框架。
Milk contains a number of beneficial fatty acids including short and medium chain and unsaturated conjugated and nonconjugated fatty acids. In this study, microRNA sequencing of mammary tissue collected in early-, peak-, mid-, and late-lactation periods was performed to determine the miRNA expression profiles. miR-16a was one of the differentially expressed miRNA and was selected for in-depth functional studies pertaining to fatty acid metabolism. The mimic of miR-16a impaired fat metabolism [triacylglycerol (TAG) and cholesterol] while knock-down of miR-16a promoted fat metabolism in vitro in bovine mammary epithelial cells (BMECs). In addition, the in vitro work with BMECs also revealed that miR-16a had a negative effect on the cellular concentration of cis 9-C18:1, total C18:1, C20:1, and C22:1 and long-chain polyunsaturated fatty acids. Therefore, these data suggesting a negative effect on fatty acid metabolism extend the discovery of the key role of miR-16a in mediating adipocyte differentiation. Through a combination of bioinformatics analysis, target gene 3' UTR luciferase reporter assays, and western blotting, we identified large tumor suppressor kinase 1 (LATS1) as a target of miR-16a. Transfection of siRNA-LATS1 into BMECs led to increases in TAG, cholesterol, and cellular fatty acid concentrations, suggesting a positive role of LATS1 in mammary cell fatty acid metabolism. In summary, data suggest that miR-16a regulates biological processes associated with intracellular TAG, cholesterol, and unsaturated fatty acid synthesis through LATS1. These data provide a theoretical and experimental framework for further clarifying the regulation of lipid metabolism in mammary cells of dairy cows.