Transforming growth factor-beta-induced inhibition of myogenesis is mediated through Smad pathway and is modulated by microtubule dynamic stability

Circ Res. 2004 Mar 19;94(5):617-25. doi: 10.1161/01.RES.0000118599.25944.D5. Epub 2004 Jan 22.

Abstract

The expression of muscle-specific genes associated with myogenesis is controlled by several myogenic transcription factors, including myogenin and MEF2D. Transforming growth factor-beta (TGF-beta) has been shown to inhibit myogenesis, yet the molecular mechanisms underlying such inhibition are not known. In the present study, TGF-beta was shown to inhibit myogenin and MEF2D expression and myotube formation in C2C12 myoblasts cultured in differentiation medium in a cell density-dependent manner. Transfection of C2C12 cells with Smad7, an antagonist for TGF-beta/Smad signaling, restored the capacity of these cells to differentiate in the presence of TGF-beta or when cultured in growth medium at low confluence, conditions that hinder muscle differentiation. Moreover, nocodazole, a microtubule-destabilizing agent, enhanced the inhibition of myogenesis exerted by TGF-beta, an effect that could be restored by tubulin-polymerizing agent taxol, both of which have been shown to affect Smad-microtubule interaction and regulate TGF-beta/Smad signaling. Our results indicate that TGF-beta inhibits myogenesis, at least in part, via Smad pathway, and provide evidence that low-dose pharmacological agents taxol and nocodazole can be used as a means to modulate myogenesis without affecting cell survival.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Cell Differentiation / drug effects
  • Cells, Cultured / drug effects
  • DNA-Binding Proteins / antagonists & inhibitors
  • DNA-Binding Proteins / biosynthesis*
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / pharmacology
  • DNA-Binding Proteins / physiology*
  • Gene Expression Regulation / drug effects
  • MEF2 Transcription Factors
  • Microtubules / drug effects
  • Microtubules / physiology*
  • Microtubules / ultrastructure
  • Muscle Development / genetics*
  • Myoblasts / cytology
  • Myoblasts / drug effects*
  • Myogenic Regulatory Factors
  • Myogenin / biosynthesis*
  • Myogenin / genetics
  • Nocodazole / pharmacology
  • Paclitaxel / pharmacology
  • Recombinant Fusion Proteins / physiology
  • Signal Transduction / drug effects
  • Signal Transduction / physiology*
  • Smad2 Protein
  • Smad3 Protein
  • Smad7 Protein
  • Trans-Activators / antagonists & inhibitors
  • Trans-Activators / genetics
  • Trans-Activators / pharmacology
  • Trans-Activators / physiology*
  • Transcription Factors / biosynthesis*
  • Transcription Factors / genetics
  • Transcription, Genetic / drug effects
  • Transfection
  • Transforming Growth Factor beta / antagonists & inhibitors
  • Transforming Growth Factor beta / pharmacology
  • Transforming Growth Factor beta / physiology*
  • Transforming Growth Factor beta1

Substances

  • DNA-Binding Proteins
  • MEF2 Transcription Factors
  • Myogenic Regulatory Factors
  • Myogenin
  • Recombinant Fusion Proteins
  • Smad2 Protein
  • Smad3 Protein
  • Smad7 Protein
  • Trans-Activators
  • Transcription Factors
  • Transforming Growth Factor beta
  • Transforming Growth Factor beta1
  • Paclitaxel
  • Nocodazole