Chinese Medical Sciences Journal ›› 2013, Vol. 28 ›› Issue (3): 140-146.doi: 10.1016/S1001-9294(13)60039-5

Previous Articles     Next Articles

Expression of microRNA-29b2-c Cluster is Positively Regulated by MyoD in L6 Cells

Chang-zheng Liu1 † *, Jing-jing Li2 †, Jin-mei Su3, Tao Jiao1, Li-juan Gou3, Xiao-dong He2, Yong-sheng Chang1†*   

  1. 1 Department of Biochemistry, National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences & Peking Union Medical Col-lege, Beijing 100005, China
    2 Department of General Surgery, 3 Department of Rheumatology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100730, China
  • Received:2013-02-15 Online:2013-09-30 Published:2013-09-30
  • Contact: *Corresponding author Tel: 86-10- 69156424, E-mail: Chang-zheng Liu, cz-liu@ibms.pumc.edu.cn, Yong-sheng Chang, changy@ibms.pumc.edu.cn
  • Supported by:

    △ Supported by the National Nature Science Foundation of China (81100608 and 30901342).

Abstract:

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.

Key words: myogenesis, myoD, microRNA-29

Copyright © 2018 Chinese Academy of Medical Sciences. All right reserved.
 
www.cmsj.cams.cn
京公安备110402430088 京ICP备06002729号-1  Powered by Magtech.

Supervised by National Health & Family Plan Commission of PRC

9 Dongdan Santiao, Dongcheng district, Beijing, 100730 China

Tel: 86-10-65105897  Fax:86-10-65133074 

E-mail: cmsj@cams.cn  www.cmsj.cams.cn

Copyright © 2018 Chinese Academy of Medical Sciences

All right reserved.

京公安备110402430088  京ICP备06002729号-1