Induction of growth arrest and polycomb gene expression by reversine allows C2C12 cells to be reprogrammed to various differentiated cell types

Proteomics. 2007 Dec;7(23):4303-16. doi: 10.1002/pmic.200700636.

Abstract

Reversine is a small, cell permeable synthetic chemical that has the ability to reprogram C2C12 myogenic cells to become various differentiated cell types. However, we still do not know how reversine works or the genes and proteins involved. Hence, we have used comparative proteomic techniques to address this issue. We have identified several proteins that were associated with cell cycle progression which were downregulated by reversine. Simultaneously, there were proteins associated with the induction of growth arrest that were upregulated. Consequently, we investigated the effects of reversine on C2C12 cell growth and established that it inhibited cell growth. Reversine had little affects on cell survival. We also investigated whether expressions of the polycomb genes, polycomb repressive complex 1 (PHC1) and Ezh2, were affected by reversine. Polycomb group genes are normally involved in chromatin based gene silencing. We found that PHC1 and Ezh2 expressions were enhanced by reversine and that it correlated with the inhibition of muscle specific transcriptional factor genes, myogenin, MyoD, and Myf5. Therefore, we believe that reversine is able to reprogram C2C12 cells to various differentiated cell types by inducing cell growth arrest, and promoting PHC1 and Ezh2 expressions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adipocytes / cytology
  • Adipocytes / drug effects
  • Animals
  • Apoptosis / drug effects
  • Blotting, Western
  • Carrier Proteins / genetics
  • Cell Cycle Proteins / metabolism
  • Cell Line
  • Cell Proliferation / drug effects*
  • Cell Survival / drug effects
  • Cell Transdifferentiation / drug effects*
  • Cell Transdifferentiation / physiology
  • Collagen Type I / analysis
  • Cyclin A / metabolism
  • Cyclin-Dependent Kinase 2 / metabolism
  • Enhancer of Zeste Homolog 2 Protein
  • GPI-Linked Proteins
  • Gene Expression Regulation / drug effects*
  • Histone-Lysine N-Methyltransferase
  • Intercellular Signaling Peptides and Proteins / metabolism
  • Membrane Proteins / metabolism
  • Mice
  • Morpholines / pharmacology*
  • Myoblasts / cytology
  • Myoblasts / drug effects*
  • Myoblasts / metabolism
  • Myogenic Regulatory Factors / genetics
  • Osteoblasts / cytology
  • Osteoblasts / drug effects
  • Osteoblasts / metabolism
  • Polycomb Repressive Complex 1
  • Polycomb Repressive Complex 2
  • Polycomb-Group Proteins
  • Proteins / genetics
  • Proteome / analysis
  • Proteome / genetics
  • Proteome / metabolism
  • Purines / pharmacology*
  • Repressor Proteins / genetics*
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • Carrier Proteins
  • Cell Cycle Proteins
  • Collagen Type I
  • Cyclin A
  • GPI-Linked Proteins
  • Gas1 protein, mouse
  • Intercellular Signaling Peptides and Proteins
  • Membrane Proteins
  • Morpholines
  • Myogenic Regulatory Factors
  • Phc1 protein, mouse
  • Polycomb-Group Proteins
  • Proteins
  • Proteome
  • Purines
  • Repressor Proteins
  • growth arrest-specific protein 6
  • Enhancer of Zeste Homolog 2 Protein
  • Ezh2 protein, mouse
  • Histone-Lysine N-Methyltransferase
  • Polycomb Repressive Complex 2
  • Polycomb Repressive Complex 1
  • Cdk2 protein, mouse
  • Cyclin-Dependent Kinase 2
  • 2-(4-morpholinoanilino)-6-cyclohexylaminopurine