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An Adipocyte Cell Culture Model to Study the Impact of Protein and Micro-RNA Modulation on Adipocyte Function
Published on: May 4, 2021
Bta-miR-6517 promotes proliferation and inhibits differentiation of pre-adipocytes by targeting PFKL
Mengli Yang1, Xiaoqian Gao1, Yanfen Ma1
1Ningxia Key Laboratory of Ruminant Molecular and Cellular Breeding, School of Agriculture, Ningxia University, Yinchuan, China.
Abstract:
The proliferation and differentiation of pre-adipocytes are regulated by microRNAs (miRNAs) and other factors. In this study, the potential functions of bta-miR-6517 in the regulation of pre-adipocyte proliferation and differentiation were explored. The qRT-PCR, oil red O staining and CCK-8 assay were used to evaluate the role of bta-miR-6517. Further, the target gene of bta-miR-6517 was identified using bioinformatics analysis, dual-luciferase reporter system and qRT-PCR system. The results found that the overexpression of bta-miR-6517 promoted the expression of proliferation marker genes and substantially increased the adipocyte proliferation vitality in the CCK-8 assay, whereas suppressing of bta-miR-6517 had the opposite effect. Overexpression bta-miR-6517 suppressed the expression of adipogenic genes, which inhibited lipid accumulation, whereas suppressing of bta-miR-6517 had the opposite effect. Furthermore, the dual-fluorescent reporter experiment results demonstrated that bta-miR-6517 directly targeted phosphofructokinase, liver type (PFKL). When bta-miR-6517 was either overexpressed or suppressed, it negatively regulated PFKL. In conclusion, we observed that bta-miR-6517 promoted adipocyte proliferation and inhibited differentiation by targeting PFKL.
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