CircHdgfrp3 Targets miR-3089-3p/miR-760-5p to Inhibit Adipocyte Differentiation
YANG Sichen1,2, WU Jiaxuan2, SHI Qiwen1, ZHANG Jin2,3*, WU Wenjing2*
The aim of this study was to determine the effect of circHdgfrp3 on adipogenic differentiation of adipocytes and its mechanism. Firstly, the results of RNA-Seq sequencing were analyzed. And circHdgfl3, a circu lar RNA molecule whose expression levels in intramuscular fat was significantly higher than that of subcutaneous fat in Large White pigs, was screened out, and the sequence similarity between pigs and mice reaches 93.87% at the genomic level. Using mouse preadipocyte line 3T3-L1 as material, circHdgfrp3 was identified, localized and ex pressed by RNase R digestion, karyotoplasmic localization and qPCR. The results showed that circHdgfrp3 existed in the cytoplasm with stable ring structure. During 3T3-L1 cell differentiation, its expression level increased first and then decreased. Transfection of si-circHdgfrp3 or pcDNA3.1-circHdgfrp3 changed its expression level during 3T3-L1 cell differentiation. It was found that circHdgfrp3 could significantly inhibit intracellular lipid droplet accumulation (P<0.05) and the expression levels of lipid formation marker genes PPARγ, C/EBPα and LPL (P<0.05) by oil red O staining and Western blot. Finally, using luciferase reporting analysis and other techniques, it was deter mined that circHdgfrp3 had a targeted binding relationship with miR-3089-3p and miR-760-5p, and could signifi cantly reduce AKT phosphorylation. In summary, circHdgfrp3 may down-regulate AKT-mTOR activity by targeting miR-3089-3p and miR-760-5p, thereby inhibiting adipogenic differentiation of adipocytes.
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