miR-30e-5p and miR-15a Synergistically Regulate Fatty Acid Metabolism in Goat Mammary Epithelial Cells via LRP6 and YAP1

被引:40
作者
Chen, Zhi [1 ]
Qiu, Huiling [2 ]
Ma, Livan [2 ]
Luo, Jun [1 ]
Sun, Shuang [1 ]
Kang, Kang [2 ]
Gou, Deming [2 ]
Loor, Juan J. [3 ]
机构
[1] Northwest A&F Univ, Coll Anim Sci & Technol, Shanxi Key Lab Mol Biol Agr, Yangling 712100, Shaanxi, Peoples R China
[2] Shenzhen Univ, Coll Life Sci, Shenzhen 518060, Peoples R China
[3] Univ Illinois, Mammalian NutriPhysioGen, Dept Anim Sci, Div Nutr Sci, Urbana, IL 61801 USA
基金
中国国家自然科学基金;
关键词
miR-30e-5p; miR-15a; fat metabolism; LRP6; YAP1; REAL-TIME PCR; MICRORNA EXPRESSION; GENE NETWORKS; LIPID-METABOLISM; ADIPOSE-TISSUE; PROTEIN; GLAND; IDENTIFICATION; MILK; DIFFERENTIATION;
D O I
10.3390/ijms17111909
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
MicroRNA (miRNA) regulates the expression of genes and influences a series of biological processes, including fatty acid metabolism. We screened the expression of miRNA in goat mammary glands during peak-lactation and non-lactating ("dry") periods, and performed an in vitro study with goat mammary epithelial cells (GMEC) prior to sequencing analysis. Results illustrated that miR-30e-5p and miR-15a were highly expressed. Utilizing a luciferase reporter assay and Western blot, low-density lipoprotein receptor-related protein 6 (LRP6) and Yes associated protein 1 (YAP1) genes were demonstrated to be a target of miR-30e-5p and miR-15a in GMEC. Moreover, we demonstrated that the overexpression of miR-30e-5p and miR-15a in GMEC promoted fat metabolism while their knockdown impaired fat metabolism. These findings extend the discovery of a key role of miR-30e-5p and miR-15a in mediating adipocyte differentiation by suggesting a role in promoting milk fat synthesis. In conclusion, our findings indicate that miR-30e-5p, together with miR-15a, represses expression of LRP6 and promotes fat metabolism in GMEC. The data expanded our knowledge on the function of miRNAs in milk fat metabolism and synthesis in ruminant mammary cells.
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页数:17
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共 52 条
[1]   Decreased expression of key tumour suppressor microRNAs is associated with lymph node metastases in triple negative breast cancer [J].
Avery-Kiejda, Kelly A. ;
Braye, Stephen G. ;
Mathe, Andrea ;
Forbes, John F. ;
Scott, Rodney J. .
BMC CANCER, 2014, 14
[2]   Characterisation of microRNA expression in post-natal mouse mammary gland development [J].
Avril-Sassen, Stefanie ;
Goldstein, Leonard D. ;
Stingl, John ;
Blenkiron, Cherie ;
Le Quesne, John ;
Spiteri, Inmaculada ;
Karagavriilidou, Konstantina ;
Watson, Christine J. ;
Tavare, Simon ;
Miska, Eric A. ;
Caldas, Carlos .
BMC GENOMICS, 2009, 10
[3]   miR-15a and miR-16 Are Implicated in Cell Cycle Regulation in a Rb-Dependent Manner and Are Frequently Deleted or Down-regulated in Non-Small Cell Lung Cancer [J].
Bandi, Nora ;
Zbinden, Samuel ;
Gugger, Mathias ;
Arnold, Marlene ;
Kocher, Verena ;
Hasan, Lara ;
Kappeler, Andreas ;
Brunner, Thomas ;
Vassella, Erik .
CANCER RESEARCH, 2009, 69 (13) :5553-5559
[4]   Akt phosphorylates the Yes-associated protein, YAP, to induce interaction with 14-3-3 and attenuation of p73-mediated apoptosis [J].
Basu, S ;
Totty, NF ;
Irwin, MS ;
Sudol, M ;
Downward, J .
MOLECULAR CELL, 2003, 11 (01) :11-23
[5]   MiR-15a and MiR-16 Control Bmi-1 Expression in Ovarian Cancer [J].
Bhattacharya, Resham ;
Nicoloso, Milena ;
Arvizo, Rochelle ;
Wang, Enfeng ;
Cortez, Angelica ;
Rossi, Simona ;
Calin, George A. ;
Mukherjee, Priyabrata .
CANCER RESEARCH, 2009, 69 (23) :9090-9095
[6]   Identification of reference genes for quantitative real-time PCR in the bovine mammary gland during the lactation cycle [J].
Bionaz, Massimo ;
Loor, Juan J. .
PHYSIOLOGICAL GENOMICS, 2007, 29 (03) :312-319
[7]   Gene Networks Driving Bovine Mammary Protein Synthesis During the Lactation Cycle [J].
Bionaz, Massimo ;
Loor, Juan J. .
BIOINFORMATICS AND BIOLOGY INSIGHTS, 2011, 5 :83-98
[8]   Gene networks driving bovine milk fat synthesis during the lactation cycle [J].
Bionaz, Massimo ;
Loor, Juan J. .
BMC GENOMICS, 2008, 9 (1)
[9]   Selection of reference genes for quantitative real-time PCR normalisation in adipose tissue, muscle, liver and mammary gland from ruminants [J].
Bonnet, M. ;
Bernard, L. ;
Bes, S. ;
Leroux, C. .
ANIMAL, 2013, 7 (08) :1344-1353
[10]   Identification and characterization of microRNA sequences from bovine mammary epithelial cells [J].
Bu, D. P. ;
Nan, X. M. ;
Wang, F. ;
Loor, J. J. ;
Wang, J. Q. .
JOURNAL OF DAIRY SCIENCE, 2015, 98 (03) :1696-1705