IkappaBalpha regulates Hes1 in osteoclast differentiation and resorption

Front Biosci (Elite Ed). 2010 Jun 1;2:1065-72. doi: 10.2741/e164.

Abstract

During osteoclast differentiation and resorption, both NF-kappaB and Notch signalling are activated. This study defines the mechanism about the influence of NFkappaB on Notch. To this end, IkappaBalphaM and Wild-type-IkappaBalpha were transfected into RAW 264.7 cells. The number of cells that differentiated into osteoclasts was quantified. The resorption area was measured. NF-kappaB transcriptional activity was determined by EMSA. Hes1, DC-STAMP and MMP-9 mRNA expression levels were determined by RT-PCR. Hes1 protein expression was determined by western blots. ChIP was used to study binding of IkappaBalpha to the Hes1 promoter. NF-kappaB inactivation inhibited the differentiation and resorption ability of osteoclasts. Compared with Wild-type cells, NF-kappaB inactivation resulted in an up-regulation of Hes1 expression, and a down-regulation of DC-STAMP and MMP-9 expression. Moreover, in response to RANKL, NF-kappaB inactivation resulted in a down-regulation of DC-STAMP and MMP-9 expression compared with Wild-type cells. The Hes1 promoter was detected by ChIP using IkappaBalpha antibody. In conclusion, our data suggest IkappaBalpha regulates Hes1-mediated activity in osteoclast differentiation and resorption, which supports a cross-talk between NF-kappaB and Notch in osteoclast activity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Basic Helix-Loop-Helix Transcription Factors / physiology*
  • Bone Resorption*
  • Cell Differentiation / physiology*
  • Cell Line
  • Cells, Cultured
  • Chromatin Immunoprecipitation
  • DNA Primers
  • Electrophoretic Mobility Shift Assay
  • Homeodomain Proteins / physiology*
  • Humans
  • I-kappa B Proteins / physiology*
  • Mice
  • NF-KappaB Inhibitor alpha
  • Osteoclasts / cytology*
  • Polymerase Chain Reaction
  • Transcription Factor HES-1

Substances

  • Basic Helix-Loop-Helix Transcription Factors
  • DNA Primers
  • Homeodomain Proteins
  • I-kappa B Proteins
  • NFKBIA protein, human
  • Nfkbia protein, mouse
  • Transcription Factor HES-1
  • NF-KappaB Inhibitor alpha
  • HES1 protein, human