title: MicroRNA-148a promotes myogenic differentiation by targeting the ROCK1 gene.
abstract: MicroRNAs are evolutionarily conserved small RNAs that post-transcriptionally regulate gene expression and have emerged as critical regulators of skeletal muscle development. Here, we identified miR-148a as a novel myogenic microRNA that mediated myogenic differentiation. The expression levels of miR-148a increased during C2C12 myoblast differentiation. Overexpression of miR-148a significantly promoted myogenic differentiation of both C2C12 myoblast and primary muscle cells. Blocking the function of miR-148a with a 2'-O-methylated antisense oligonucleotide inhibitor repressed C2C12 myoblast differentiation. Using a bioinformatics approach, we identified Rho-associated coiled-coil containing protein kinase 1 (ROCK1), a known inhibitor of myogenesis, as a target of miR-148a. A dual-luciferase reporter assay was used to demonstrate that miR-148a directly targeted the 3'-UTR of ROCK1. In addition, the overexpression of miR-148a decreased the protein expression of ROCK1 in C2C12 myoblast and primary muscle cells. Furthermore, ROCK1 inhibition with specific siRNA leaded to accelerated myogenic differentiation progression, underscoring a negative regulatory function of ROCK1 in myogenesis. Therefore, our results revealed a novel mechanism in which miR-148a positively regulates myogenic differentiation via ROCK1 down-regulation.
authors.affiliation: State Key Laboratory for Animal Nutrition, The Key Laboratory for Domestic Animal Genetic Resources and Breeding of Ministry of Agriculture of China, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, PR China.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhang
authors.forename: Zheng-zhou
authors.initials: ZZ
authors.lastname: Ying
authors.forename: Zhong-lin
authors.initials: ZL
authors.lastname: Tang
authors.forename: Liang-qi
authors.initials: LQ
authors.lastname: Long
authors.forename: Kui
authors.initials: K
authors.lastname: Li
mesh_terms: D020413:3' Untranslated Regions / Q000502:physiology; D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D015536:Down-Regulation / Q000502:physiology*; D051379:Mice; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D024510:Muscle Development / Q000502:physiology*; D032448:Myoblasts, Skeletal / Q000166:cytology / Q000378:metabolism*; D054460:rho-Associated Kinases / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D020413:3' Untranslated Regions; D035683:MicroRNAs; C554644:Mirn148 microRNA, mouse; C515854:Rock1 protein, mouse; D054460:rho-Associated Kinases