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miR-411 is up-regulated in FSHD myoblasts and suppresses myogenic factors
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)...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3637251/ https://www.ncbi.nlm.nih.gov/pubmed/23561550 http://dx.doi.org/10.1186/1750-1172-8-55 |
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author | Harafuji, Naoe Schneiderat, Peter Walter, Maggie C Chen, Yi-Wen |
author_facet | Harafuji, Naoe Schneiderat, Peter Walter, Maggie C Chen, Yi-Wen |
author_sort | Harafuji, Naoe |
collection | PubMed |
description | 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 C(2)C(12) 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). C(2)C(12) 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. |
format | Online Article Text |
id | pubmed-3637251 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-36372512013-04-27 miR-411 is up-regulated in FSHD myoblasts and suppresses myogenic factors Harafuji, Naoe Schneiderat, Peter Walter, Maggie C Chen, Yi-Wen Orphanet J Rare Dis Research 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 C(2)C(12) 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). C(2)C(12) 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. BioMed Central 2013-04-05 /pmc/articles/PMC3637251/ /pubmed/23561550 http://dx.doi.org/10.1186/1750-1172-8-55 Text en Copyright © 2013 Harafuji et al.; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Harafuji, Naoe Schneiderat, Peter Walter, Maggie C Chen, Yi-Wen miR-411 is up-regulated in FSHD myoblasts and suppresses myogenic factors |
title | miR-411 is up-regulated in FSHD myoblasts and suppresses myogenic factors |
title_full | miR-411 is up-regulated in FSHD myoblasts and suppresses myogenic factors |
title_fullStr | miR-411 is up-regulated in FSHD myoblasts and suppresses myogenic factors |
title_full_unstemmed | miR-411 is up-regulated in FSHD myoblasts and suppresses myogenic factors |
title_short | miR-411 is up-regulated in FSHD myoblasts and suppresses myogenic factors |
title_sort | mir-411 is up-regulated in fshd myoblasts and suppresses myogenic factors |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3637251/ https://www.ncbi.nlm.nih.gov/pubmed/23561550 http://dx.doi.org/10.1186/1750-1172-8-55 |
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