Vitamin K induces osteoblast differentiation through pregnane X receptor-mediated transcriptional control of the Msx2 gene

Mol Cell Biol. 2007 Nov;27(22):7947-54. doi: 10.1128/MCB.00813-07. Epub 2007 Sep 17.

Abstract

Vitamin K is a fat-soluble vitamin that serves as a coenzyme for vitamin K-dependent carboxylase. Besides its canonical action, vitamin K binds to the steroid and xenobiotic receptor (SXR)/pregnane X receptor (PXR) and modulates gene transcription. To determine if the osteoprotective action of vitamin K is the result of the PXR/SXR pathway, we screened by two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis the PXR/SXR target genes in an osteoblastic cell line (MC3T3-E1) treated with a vitamin K2 (menaquinone 4 [MK4]). Osteoblastic differentiation of MC3T3-E1 cells was induced by MK4. Msx2, an osteoblastogenic transcription factor, was identified as an MK4-induced gene. Functional analysis of the Msx2 gene promoter mapped a vitamin K-responsive element (PXR-responsive element [PXRE]) that was directly bound by a PXR/retinoid X receptor alpha heterodimer. In a chromatin immunoprecipitation analysis, PXR was recruited together with a coactivator, p300, to the PXRE in the Msx2 promoter. MK4-bound PXR cooperated with estrogen-bound estrogen receptor alpha to control transcription at the Msx2 promoter. Knockdown of either PXR or Msx2 attenuated the effect of MK4 on osteoblastic differentiation. Thus, the present study suggests that Msx2 is a target gene for PXR activated by vitamin K and suggests that the osteoprotective action of MK4 in the human mediates, at least in part, a genomic pathway of vitamin K signaling.

Publication types

  • Research Support, Non-U.S. Gov't
  • Retracted Publication

MeSH terms

  • Animals
  • Biomarkers / metabolism
  • Cell Differentiation / drug effects*
  • Cell Differentiation / physiology
  • Cells, Cultured
  • DNA-Binding Proteins* / genetics
  • DNA-Binding Proteins* / metabolism
  • Estradiol / metabolism
  • Estradiol / pharmacology
  • Estrogen Receptor alpha / genetics
  • Estrogen Receptor alpha / metabolism
  • Gene Expression Regulation*
  • Hemostatics / metabolism
  • Hemostatics / pharmacology
  • Homeodomain Proteins* / genetics
  • Homeodomain Proteins* / metabolism
  • Humans
  • Mice
  • Mice, Knockout
  • Osteoblasts / cytology
  • Osteoblasts / drug effects*
  • Osteoblasts / physiology*
  • Pregnane X Receptor
  • Promoter Regions, Genetic
  • Receptors, Steroid / genetics
  • Receptors, Steroid / metabolism*
  • Response Elements
  • Retinoid X Receptor alpha / genetics
  • Retinoid X Receptor alpha / metabolism
  • Signal Transduction / physiology
  • Transcription, Genetic
  • Vitamin K / metabolism
  • Vitamin K / pharmacology*
  • Vitamin K 2 / analogs & derivatives
  • Vitamin K 2 / metabolism
  • Vitamin K 2 / pharmacology

Substances

  • Biomarkers
  • DNA-Binding Proteins
  • Estrogen Receptor alpha
  • Hemostatics
  • Homeodomain Proteins
  • MSX2 protein
  • Pregnane X Receptor
  • Receptors, Steroid
  • Retinoid X Receptor alpha
  • Vitamin K 2
  • Vitamin K
  • menatetrenone
  • Estradiol