title: miR-411 is up-regulated in FSHD myoblasts and suppresses myogenic factors.
abstract: BACKGROUND
  Facioscapulohumeral muscular dystrophy (FSHD) is an autosomal dominant muscle disorder, which is linked to the contraction of the D4Z4 array at chromosome 4q35. Recent studies suggest that this shortening of the D4Z4 array leads to aberrant expression of double homeobox protein 4 (DUX4) and causes FSHD. In addition, misregulation of microRNAs (miRNAs) has been reported in muscular dystrophies including FSHD. In this study, we identified a miRNA that is differentially expressed in FSHD myoblasts and investigated its function.
  
  
  METHODS
  To identify misregulated miRNAs and their potential targets in FSHD myoblasts, we performed expression profiling of both miRNA and mRNA using TaqMan Human MicroRNA Arrays and Affymetrix Human Genome U133A plus 2.0 microarrays, respectively. In addition, we over-expressed miR-411 in C₂C₁₂ cells to determine the effect of miR-411 on myogenic markers.
  
  
  RESULTS
  Using miRNA and mRNA expression profiling, we identified 8 miRNAs and 1,502 transcripts that were differentially expressed in FSHD myoblasts during cell proliferation. One of the 8 differentially expressed miRNAs, miR-411, was validated by quantitative RT-PCR in both primary (2.1 fold, p<0.01) and immortalized (2.7 fold, p<0.01) myoblasts. In situ hybridization showed cytoplasmic localization of miR-411 in FSHD myoblasts. By analyzing both miRNA and mRNA data using Partek Genomics Suite, we identified 4 mRNAs potentially regulated by miR-411 including YY1 associated factor 2 (YAF2). The down-regulation of YAF2 in immortalized myoblasts was validated by immunoblotting (-3.7 fold, p<0.01). C₂C₁₂ cells were transfected with miR-411 to determine whether miR-411 affects YAF2 expression in myoblasts. The results showed that over-expression of miR-411 reduced YAF2 mRNA expression. In addition, expression of myogenic markers including Myod, myogenin, and myosin heavy chain 1 (Myh1) were suppressed by miR-411.
  
  
  CONCLUSIONS
  The study demonstrated that miR-411 was differentially expressed in FSHD myoblasts and may play a role in regulating myogenesis.
authors.affiliation: Center for Genetic Medicine Research, Children's Research Institute, Washington, DC, USA.
authors.forename: Naoe
authors.initials: N
authors.lastname: Harafuji
authors.forename: Peter
authors.initials: P
authors.lastname: Schneiderat
authors.forename: Maggie C
authors.initials: MC
authors.lastname: Walter
authors.forename: Yi-Wen
authors.initials: YW
authors.lastname: Chen
mesh_terms: D002461:Cell Line, Transformed; D002894:Chromosomes, Human, Pair 4; D020869:Gene Expression Profiling; D006801:Humans; D017403:In Situ Hybridization; D035683:MicroRNAs / Q000235:genetics*; D009124:Muscle Proteins / Q000235:genetics; D020391:Muscular Dystrophy, Facioscapulohumeral / Q000235:genetics* / Q000473:pathology; D017570:MyoD Protein / Q000235:genetics; D032446:Myoblasts / Q000378:metabolism*; D018008:Myogenin / Q000235:genetics; D012097:Repressor Proteins / Q000235:genetics; D015854:Up-Regulation
chemical_list: C582326:MIRN411 microRNA, human; D035683:MicroRNAs; D009124:Muscle Proteins; D017570:MyoD Protein; D018008:Myogenin; D012097:Repressor Proteins; C104531:YAF2 protein, human
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