MiR-21-5p promotes differentiation and reduces oleic acid-induced lipid droplet accumulation in C2C12 myoblasts by targeting FBXO11

¹College of Animal Science, Jilin University, China, Changchun, China
²College of Life Science, Baicheng Normal University, Baicheng 137000, China, Baicheng, China

Correspondence: Changhong Li,Email: 13331750780@163.com
Linlin Hao,Email: 13331750780@163.com

Received: 24 September 2024 • Revised: 24 November 2024 • Accepted: 24 September 2024

Abstract

Objective
This study aimed to investigate the function of miRNA 21-5p in regulating the differentiation of C2C12 myoblasts and intramuscular lipid droplet accumulation in myotubes.
Methods
The role of miR-21-5p in the proliferation and differentiation of myofibroblasts and intracellular lipid accumulation was analyzed using bioinformatics, CCK-8 assay, RT-qPCR, immunoblotting, immunofluorescence staining, and Oil Red O staining.
Results
Analysis of porcine BodyMap transcriptomic data revealed differential expression of miRNA 21-5p in skeletal muscle and adipose tissue. Bioinformatics analysis combined with a dual-luciferase reporter assay demonstrated that FBXO11 serves as a direct target of miR-21-5p. Transfection experiments involving a miR-21-5p mimic, miR-21-5p inhibitor, and si-FBXO11 in C2C12 cells showed that overexpression of miR-21-5p or silencing of FBXO11 significantly enhanced the proliferation of C2C12 cells, upregulated myogenesis-related factors, and promoted myotube formation. Additionally, oleic acid-induced lipid accumulation in myotubes was suppressed, accompanied by reduced expression of adipogenesis-related genes. Conversely, inhibition of miR-21-5p expression produced opposite effects.
Conclusion
These findings indicate that miR-21-5p promotes proliferation and differentiation while inhibiting intramyocellular lipid deposition by targeting the 3'-untranslated region (3’UTR) of FBXO11 in myogenic cell. The results suggest that miR-21-5p could serve as a potential miRNA biomarker for regulating intramuscular adipogenesis.

Keywords: miR-21-5p; FBXO11; C2C12 myoblast; proliferation; differentiation; intramuscular lipid deposition