ERβ induces the differentiation of cultured osteoblasts by both Wnt/β-catenin signaling pathway and estrogen signaling pathways

Exp Cell Res. 2015 Jul 1;335(1):107-14. doi: 10.1016/j.yexcr.2015.04.020. Epub 2015 May 7.

Abstract

Although 17β-estradial (E2) is known to stimulate bone formation, the underlying mechanisms are not fully understood. Recent studies have implicated the Wnt/β-catenin pathway as a major signaling cascade in bone biology. The interactions between Wnt/β-catenin signaling pathway and estrogen signaling pathways have been reported in many tissues. In this study, E2 significantly increased the expression of β-catenin by inducing phosphorylations of GSK3β at serine 9. ERβ siRNAs were transfected into MC3T3-E1 cells and revealed that ERβ involved E2-induced osteoblasts proliferation and differentiation via Wnt/β-catenin signaling. The osteoblast differentiation genes (BGP, ALP and OPN) and proliferation related gene (cyclin D1) expression were significantly induced by E2-mediated ERβ. Furthermore immunofluorescence and immunoprecipitation analysis demonstrated that E2 induced the accumulation of β-catenin protein in the nucleus which leads to interaction with T-cell-specific transcription factor/lymphoid enhancer binding factor (TCF/LEF) transcription factors. Taken together, these findings suggest that E2 promotes osteoblastic proliferation and differentiation by inducing proliferation-related and differentiation-related gene expression via ERβ/GSK-3β-dependent Wnt/β-catenin signaling pathway. Our findings provide novel insights into the mechanisms of action of E2 in osteoblastogenesis.

Keywords: 17β-estradial (E2); ERβ; GSK3β; Osteoblast; Wnt/β-catenin signaling; β-catenin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3 Cells
  • Adaptor Proteins, Signal Transducing / biosynthesis
  • Animals
  • Carcinoembryonic Antigen / biosynthesis
  • Cell Differentiation
  • Cell Line
  • Cell Proliferation
  • Cyclin D1 / biosynthesis
  • Estradiol / metabolism*
  • Estradiol / pharmacology
  • Estrogen Receptor beta / genetics
  • Estrogen Receptor beta / metabolism*
  • Glycogen Synthase Kinase 3 / metabolism
  • Glycogen Synthase Kinase 3 beta
  • Membrane Proteins / biosynthesis
  • Mice
  • Osteoblasts / cytology*
  • Osteoblasts / drug effects
  • Osteoblasts / metabolism
  • Osteogenesis / genetics
  • Phosphorylation
  • RNA Interference
  • RNA, Small Interfering
  • Wnt Proteins / metabolism*
  • Wnt Signaling Pathway
  • beta Catenin / metabolism*

Substances

  • Adaptor Proteins, Signal Transducing
  • Atrnl1 protein, mouse
  • Carcinoembryonic Antigen
  • Ccnd1 protein, mouse
  • Ceacam1 protein, mouse
  • Estrogen Receptor beta
  • Membrane Proteins
  • RNA, Small Interfering
  • Wnt Proteins
  • beta Catenin
  • Cyclin D1
  • Estradiol
  • Glycogen Synthase Kinase 3 beta
  • Gsk3b protein, mouse
  • Glycogen Synthase Kinase 3