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Expression of microRNA-29b2-c Cluster is Positively Regulated by MyoD in L6 Cells.
MedLine Citation:
PMID:  24074615     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Methods The expression of myoD and miR-29 family was detected by using real-time PCR and Western blot analysis. The potential promoter and transcription factors binding sites of miR-29 clusters were predicted by Promoter scan and transcriptional factor search. The promoter sequence of miR-29b1-a and miR-29b2-c cluster was cloned into a luciferase reporter plasmid and the regulatory effect of myoD was analyzed by using dual luciferase reporter assay. Electrophoretic mobility shift assay was further conducted to indicate the binding of myoD on specific sequence. Moreover, overexpression of myoD was achieved by a recombinant adenovirus system (Ad-myoD). L6 cells were infected with Ad-myoD and real-time PCR was conducted to analyze the expression of miR-29b and miR-29c.Results The expression levels of myoD, miR-29a, miR-29b, and miR-29c were increased in L6 myoblast differentiated to myotube. The expression of myoD, miR-29b, and miR-29c was up-regulated in L6 myotube treated with glucose and insulin, but miR-29a depicted no significant change. Dual luciferase reporter gene assay showed that myoD functioned as a positive regulator of miR-29b2-c expression and myoD could bind to the specific sequence located at the promoter region of miR-29b2-c cluster. Enforced expression of myoD led to a marked increase of miR-29b and miR-29c levels in L6 cells.ConclusionMyoD might act as a crucial regulator of myogenesis and glucose metabolism in muscle through regulating the expression of miR-29b2-c.
Authors:
Chang-Zheng Liu; Jing-Jing Li; Jin-Mei Su; Tao Jiao; Li-Juan Gou; Xiao-Dong He; Yong-Sheng Chang
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Publication Detail:
Type:  JOURNAL ARTICLE    
Journal Detail:
Title:  Chinese medical sciences journal = Chung-kuo i hsueh k'o hsueh tsa chih / Chinese Academy of Medical Sciences     Volume:  28     ISSN:  1001-9294     ISO Abbreviation:  Chin. Med. Sci. J.     Publication Date:  2013 Sep 
Date Detail:
Created Date:  2013-9-30     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9112559     Medline TA:  Chin Med Sci J     Country:  -    
Other Details:
Languages:  ENG     Pagination:  140-146     Citation Subset:  -    
Affiliation:
Department of Biochemistry, National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences.
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