MiR-21-5p enhances differentiation and mitigates oleic acid-induced lipid droplet accumulation in C2C12 myoblasts by targeting FBXO11
MiR-21-5p enhances differentiation and mitigates oleic acid-induced lipid droplet accumulation in C2C12 myoblasts by targeting FBXO11
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Objective: This study aimed to investigate the role of miRNA 21-5p in regulating the differentiation of C2C12 myoblasts and intramuscular lipid droplets 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, quantitative real-time polymerase chain reaction, immunoblotting, immunofluorescence staining, and Oil Red O staining.Results: The analysis of porcine BodyMap transcriptome 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. Furthermore, 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 of FBXO11 in myogenic cell. The results suggest that miR-21-5p could serve as a potential miRNA biomarker for regulating intramuscular adipogenesis.
Abstract
Objective: This study aimed to investigate the role of miRNA 21-5p in regulating the differentiation of C2C12 myoblasts and intramuscular lipid droplets 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, quantitative real-time polymerase chain reaction, immunoblotting, immunofluorescence staining, and Oil Red O staining. Results: The analysis of porcine BodyMap transcriptome 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. Furthermore, 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 of FBXO11 in myogenic cell. The results suggest that miR-21-5p could serve as a potential miRNA biomarker for regulating intramuscular adipogenesis.