Wnt proteins prevent apoptosis of both uncommitted osteoblast progenitors and differentiated osteoblasts by beta-catenin-dependent and -independent signaling cascades involving Src/ERK and phosphatidylinositol 3-kinase/AKT

J Biol Chem. 2005 Dec 16;280(50):41342-51. doi: 10.1074/jbc.M502168200. Epub 2005 Oct 25.

Abstract

Genetic studies in humans and mice have revealed an important role of the Wnt signaling pathway in the regulation of bone mass, resulting from potent effects on the control of osteoblast progenitor proliferation, commitment, differentiation, and perhaps osteoblast apoptosis. To establish the linkage between Wnts and osteoblast survival and to elucidate the molecular pathways that link the two, we have utilized three cell models: the uncommitted bipotential C2C12 cells, the pre-osteoblastic cell line MC3T3-E1, and bone marrow-derived OB-6 osteoblasts. Serum withdrawal-induced apoptosis was prevented by the canonical Wnts (Wnt3a and Wnt1) and the noncanonical Wnt5a in all cell types. Wnt3a induced LRP5-independent transient phosphorylation and nuclear accumulation of ERKs and phosphorylation of Src and Akt. The anti-apoptotic effect of Wnt3a was abrogated by inhibitors of canonical Wnt signaling, as well as by inhibitors of MEK, Src, phosphatidylinositol 3-kinase (PI3K), or Akt kinases, or by the addition of cycloheximide to the culture medium. Wnt3a-induced phosphorylation of GSK-3beta and downstream activation of beta-catenin-mediated transcription required ERK, PI3K, and Akt signaling. Wnt3a increased the expression of the anti-apoptotic protein Bcl-2 in an ERK-dependent manner. Beta-catenin-mediated transcription was permissive for the anti-apoptotic actions of Wnt1 and Wnt3a but was dispensable for the anti-apoptotic action of Wnt5a. However, Src, ERKs, PI3K, and Akt kinases were required for the anti-apoptotic effects of Wnt5a. These results demonstrate for the first time that Wnt proteins, irrespective of their ability to stimulate canonical Wnt signaling, prolong the survival of osteoblasts and uncommitted osteoblast progenitors via activation of the Src/ERK and PI3K/Akt signaling cascades.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Apoptosis*
  • Blotting, Western
  • Cell Differentiation
  • Cell Line
  • Cell Nucleus / metabolism
  • Cell Survival
  • Culture Media / pharmacology
  • Culture Media, Serum-Free / pharmacology
  • Cycloheximide / pharmacology
  • Enzyme Inhibitors / pharmacology
  • Extracellular Signal-Regulated MAP Kinases / metabolism*
  • Gene Expression Regulation, Enzymologic*
  • Green Fluorescent Proteins / metabolism
  • Mice
  • Models, Biological
  • Osteoblasts / cytology*
  • Osteoblasts / metabolism
  • Phosphatidylinositol 3-Kinases / metabolism*
  • Phosphorylation
  • Proto-Oncogene Proteins / metabolism
  • Proto-Oncogene Proteins c-akt / metabolism*
  • RNA / metabolism
  • Signal Transduction
  • Stem Cells / cytology*
  • Time Factors
  • Transcription, Genetic
  • Transfection
  • Wnt Proteins / metabolism*
  • Wnt-5a Protein
  • Wnt1 Protein / metabolism
  • Wnt3 Protein
  • Wnt3A Protein
  • beta Catenin / metabolism*

Substances

  • Culture Media
  • Culture Media, Serum-Free
  • Enzyme Inhibitors
  • Proto-Oncogene Proteins
  • WNT3A protein, human
  • Wnt Proteins
  • Wnt-5a Protein
  • Wnt1 Protein
  • Wnt1 protein, mouse
  • Wnt3 Protein
  • Wnt3A Protein
  • Wnt3a protein, mouse
  • Wnt5a protein, mouse
  • beta Catenin
  • Green Fluorescent Proteins
  • RNA
  • Cycloheximide
  • Phosphatidylinositol 3-Kinases
  • Proto-Oncogene Proteins c-akt
  • Extracellular Signal-Regulated MAP Kinases