MicroRNA-378 regulates adipogenic differentiation in bovine intramuscular preadipocytes by targeting CaMKK2

Dongwei Li; Heng Wang; Yongmin Li; Changqing Qu; Yunhai Zhang; Hongyu Liu; Xiaorong Zhang

doi:10.1080/21623945.2021.1982526

MicroRNA-378 regulates adipogenic differentiation in bovine intramuscular preadipocytes by targeting CaMKK2

Adipocyte. 2021 Dec;10(1):483-492. doi: 10.1080/21623945.2021.1982526.

Authors

Dongwei Li^{1

2}, Heng Wang¹, Yongmin Li², Changqing Qu³, Yunhai Zhang¹, Hongyu Liu¹, Xiaorong Zhang¹

Affiliations

¹ College of Animal Science and Technology, Anhui Agricultural University, Hefei China.
² Conservation Biology Research Center, School of Biology and Food Engineering, Fuyang Normal University, Fuyang, China.
³ Anhui Provincial Engineering Technology Research Center of Anti-aging Chinese Herbal Medicine, Fuyang Normal University, Fuyang, China.

Abstract

Intramuscular fat, as one of the most important palatability attribute of beef carcase, is the primary determinant of beef quality. The research of adipogenesis mechanism would provide new insight into intramuscular fatty deposition. Here, the role of microRNA-378 was investigated during bovine adipogenic differentiation. It was revealed that miR-378 expression exists variably in bovine major tissue and organs by RT-qPCR. It was predicted that miR-378 targets CaMKK2, as an AMPKα kinase, by DIANA Tools. For better research, primary preadipocytes with stable transfection for up-/down-regulated expression of miR-378 were constructed by lentiviral vectors with GFP gene. The analyses of qPCR showed that PPARγ and adiponectin mRNA levels increased, but C/EBPβ, pref-1 and CaMKK2 mRNA levels decreased during adipogenic differentiation. When miR-378 was overexpressed, preadipocytes proliferation became slower, there are more cellular lipid droplets, and PPARγ and C/EBPβ mRNA levels were higher, but pref-1, adiponectin and CaMKK2 were lower than control groups. Luciferase assay and western blot analysis validated that miR-378 binds the nucleotide sites of the 3'- untranslated region of CaMKK2, which inhibits the mRNA and protein expression of CaMKK2. These findings suggest that miR-378 promotes adipogenic differentiation in bovine intramuscular preadipocytes by targeting CaMKK2 via AMPK signalling pathway.

Keywords: Camkk2; Microrna-378; adipogenic differentiation; bovine; intramuscular preadipocytes.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adipocytes
Adipogenesis* / genetics
Animals
Cattle
Cell Differentiation
MicroRNAs* / genetics
PPAR gamma

Substances

MicroRNAs
PPAR gamma

Grants and funding

This work was funded by the National Natural Science Foundation of China (31101696), Program of National Beef Cattle and Yak Industrial Technology System (CARS-37), Anhui Province Higher School Natural Science Key Program of China (KJ2018A0350), Anhui Province Science and Technology Major Special Project (202003a06020020, 201904f06020018), Scientific Research Project of Cooperation between Fuyang Municipal Government and Fuyang Normal University (XDHX201736), and Ph.D. Research Startup Fund of Fuyang Normal University (2016);