MicroRNA-200a regulates Grb2 and suppresses differentiation of mouse embryonic stem cells into endoderm and mesoderm

PLoS One. 2013 Jul 18;8(7):e68990. doi: 10.1371/journal.pone.0068990. Print 2013.

Abstract

The mechanisms by which microRNAs (miRNAs) affect cell fate decisions remain poorly understood. Herein, we report that miR-200a can suppress the differentiation of mouse embryonic stem (ES) cells into endoderm and mesoderm. Interestingly, miR-200a directly targets growth factor receptor-bound protein 2 (Grb2), which is a key adaptor in the Erk signaling pathway. Furthermore, high levels of miR-200a dramatically decrease Grb2 levels and suppress the appearance of mesoderm and endoderm lineages in embryoid body formation, as well as suppressing the activation of Erk. Finally, Grb2 supplementation significantly rescues the miR-200a-induced layer-formation bias and the Erk suppression. Collectively, our results demonstrate that miR-200a plays critical roles in ES cell lineage commitment by directly regulating Grb2 expression and Erk signaling.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkaline Phosphatase
  • Animals
  • Blotting, Western
  • Cell Differentiation / physiology*
  • Embryonic Stem Cells / cytology*
  • Endoderm / cytology*
  • Fluorescent Antibody Technique
  • GRB2 Adaptor Protein / metabolism*
  • Luciferases
  • Mesoderm / cytology*
  • Mice
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Microscopy, Fluorescence
  • Oligonucleotides / genetics
  • Real-Time Polymerase Chain Reaction

Substances

  • GRB2 Adaptor Protein
  • MicroRNAs
  • Mirn200 microRNA, mouse
  • Oligonucleotides
  • Luciferases
  • Alkaline Phosphatase

Grants and funding

This work was supported by grants from the Ministry of Science and Technology (2011CB965100, 2011DFA30480, 2010CB944900, 2010CB945000, 2012CB966603, 2011CBA01100, 2013CB967401 and 2013CB531606), the National Natural Science Foundation of China (31210103905, 91219305, 31201107, 31101061, 81170499, 31071306, 31000378, 31171432, and 81273282), the Science and Technology Commission of Shanghai Municipality (12ZR1450900, 11ZR1438500, 11XD1405300 and 11JC1410902), and IRT1168 and 20110072110039 from Ministry of Education. The work was also supported by Fundamental Research Funds for the Central Universities (2000219066, 2000219067 and 2000219077). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.