MiR-16a Regulates Milk Fat Metabolism by Targeting Large Tumor Suppressor Kinase 1 (LATS1) in Bovine Mammary Epithelial Cells

J Agric Food Chem. 2019 Oct 9;67(40):11167-11178. doi: 10.1021/acs.jafc.9b04883. Epub 2019 Sep 30.

Abstract

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.

Keywords: LATS1; dairy cow; miR-16a; unsaturated fatty acid.

MeSH terms

  • Animals
  • Cattle / genetics
  • Cattle / metabolism*
  • Cholesterol / metabolism
  • Epithelial Cells / enzymology*
  • Epithelial Cells / metabolism
  • Fatty Acids / metabolism
  • Female
  • Gene Expression Regulation
  • Lipid Metabolism*
  • Mammary Glands, Animal / enzymology*
  • Mammary Glands, Animal / metabolism
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Milk / metabolism*
  • Protein Serine-Threonine Kinases / genetics
  • Protein Serine-Threonine Kinases / metabolism*
  • Triglycerides / metabolism

Substances

  • Fatty Acids
  • MicroRNAs
  • Triglycerides
  • Cholesterol
  • Protein Serine-Threonine Kinases