JunB as a potential mediator of PTHrP actions: new gene targets Ephrin B1 and VCAM-1

Oral Dis. 2008 Nov;14(8):713-26. doi: 10.1111/j.1601-0825.2008.01489.x.

Abstract

Parathyroid hormone-related protein (PTHrP) is an integral mediator of physiologic and pathologic processes and has demonstrated actions in the periodontium. PTHrP functions via AP-1, and specifically through JunB. This study identified JunB-dependent downstream mediators of PTHrP using OCCM cementoblastic transfectants with JunB over- or reduced expression. Over-expressing cells showed an increase in proliferation, while the opposite was seen in siRNA transfected cells. Microarray analysis of over-expressing cells revealed more than 1000 regulated genes. Three genes were investigated in more detail. The PTH/PTHrP receptor (PTHR1) and ephrin B1 (EfnB1) were down-regulated, and vascular cell adhesion molecule-1 (VCAM-1) was up-regulated with JunB over-expression. JunB siRNA transfectants had increased PTHR1, but reduced ephrin B1 and unaltered VCAM-1 in vitro. To validate these targets, parental OCCM cells and primary osteoblasts were treated with PTHrP, resulting in reduced PTHR1 and ephrin B1, and increased VCAM-1. Cell transfectants were implanted subcutaneously in vivo, and microarray analysis and RT-PCR performed. Over-expression of JunB down-regulated PTHR1 and ephrin B1, and increased VCAM-1. JunB siRNA transfectant implants had increased PTHR1 and ephrin B1, but no altered VCAM-1. These data highlight new gene targets for PTHrP and indicate JunB is a critical mediator of PTHrP actions.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Apoptosis / genetics
  • Cell Count
  • Cell Line
  • Cell Proliferation
  • Dental Cementum / pathology
  • Down-Regulation / genetics
  • Ephrin-B1 / genetics*
  • Gene Expression Regulation / genetics
  • Mice
  • Mice, Nude
  • Osteoblasts / pathology
  • Osteoblasts / transplantation
  • Parathyroid Hormone-Related Protein / genetics*
  • Protein Array Analysis
  • Proto-Oncogene Proteins c-jun / genetics*
  • RNA, Small Interfering / genetics
  • Receptor, Parathyroid Hormone, Type 1 / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • Subcutaneous Tissue / pathology
  • Transcription Factor AP-1 / genetics
  • Transfection
  • Up-Regulation / genetics
  • Vascular Cell Adhesion Molecule-1 / genetics*

Substances

  • Efnb1 protein, mouse
  • Ephrin-B1
  • Parathyroid Hormone-Related Protein
  • Proto-Oncogene Proteins c-jun
  • RNA, Small Interfering
  • Receptor, Parathyroid Hormone, Type 1
  • Transcription Factor AP-1
  • Vascular Cell Adhesion Molecule-1