Yi Li, Jiayuan Fang, Yingying Jiao, Qinchuan Lv, Xingyu Xiao, Shuo Zheng, Xi Chen, Jie Song, Xunming Zhang, Libo Zhang, Ze Ma, Changhong Li, Linlin Hao
{"title":"MiR-21-5p通过靶向FBXO11促进C2C12成肌细胞分化并减少油酸诱导的脂滴积聚。","authors":"Yi Li, Jiayuan Fang, Yingying Jiao, Qinchuan Lv, Xingyu Xiao, Shuo Zheng, Xi Chen, Jie Song, Xunming Zhang, Libo Zhang, Ze Ma, Changhong Li, Linlin Hao","doi":"10.5713/ab.24.0665","DOIUrl":null,"url":null,"abstract":"<p><strong>Objective: </strong>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.</p><p><strong>Methods: </strong>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.</p><p><strong>Results: </strong>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.</p><p><strong>Conclusion: </strong>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.</p>","PeriodicalId":7825,"journal":{"name":"Animal Bioscience","volume":" ","pages":"1279-1290"},"PeriodicalIF":2.4000,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12061579/pdf/","citationCount":"0","resultStr":"{\"title\":\"MiR-21-5p enhances differentiation and mitigates oleic acid-induced lipid droplet accumulation in C2C12 myoblasts by targeting FBXO11.\",\"authors\":\"Yi Li, Jiayuan Fang, Yingying Jiao, Qinchuan Lv, Xingyu Xiao, Shuo Zheng, Xi Chen, Jie Song, Xunming Zhang, Libo Zhang, Ze Ma, Changhong Li, Linlin Hao\",\"doi\":\"10.5713/ab.24.0665\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Objective: </strong>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.</p><p><strong>Methods: </strong>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.</p><p><strong>Results: </strong>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.</p><p><strong>Conclusion: </strong>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.</p>\",\"PeriodicalId\":7825,\"journal\":{\"name\":\"Animal Bioscience\",\"volume\":\" \",\"pages\":\"1279-1290\"},\"PeriodicalIF\":2.4000,\"publicationDate\":\"2025-06-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12061579/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Animal Bioscience\",\"FirstCategoryId\":\"97\",\"ListUrlMain\":\"https://doi.org/10.5713/ab.24.0665\",\"RegionNum\":2,\"RegionCategory\":\"农林科学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2025/2/27 0:00:00\",\"PubModel\":\"Epub\",\"JCR\":\"Q1\",\"JCRName\":\"AGRICULTURE, DAIRY & ANIMAL SCIENCE\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Animal Bioscience","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.5713/ab.24.0665","RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/2/27 0:00:00","PubModel":"Epub","JCR":"Q1","JCRName":"AGRICULTURE, DAIRY & ANIMAL SCIENCE","Score":null,"Total":0}
MiR-21-5p enhances differentiation and mitigates oleic acid-induced lipid droplet accumulation in C2C12 myoblasts by targeting FBXO11.
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.