Endothelin-1 enhances cell migration through COX-2 up-regulation in human chondrosarcoma

Biochim Biophys Acta. 2013 Jun;1830(6):3355-64. doi: 10.1016/j.bbagen.2013.03.014. Epub 2013 Mar 21.

Abstract

Background: Chondrosarcoma is a type of highly malignant tumor with a potent capacity of local invasion and distant metastasis. The effect of endothelin-1 (ET-1) on migration activity in human chondrosarcoma cells is not clearly understood. Here, we found that ET-1 increased the migration and expression of cyclooxygenase (COX)-2 in human chondrosarcoma cells.

Methods: ET-1-mediated COX-2 expression was assessed by qPCR and Western blot analysis. The mechanisms of action of ET-1 in different signaling pathways were studied using Western blotting. Knockdown of proteins was achieved by transfection with siRNA. Chromatin immunoprecipitation assays were used to study in vivo binding of c-Jun to the COX-2 promoter.

Results: Human chondrosarcoma tissues had significant expression levels of ET-1 and COX-2, which were higher than that in normal cartilage. Exogenous ET-1 increased cell migration and the expression of COX-2. In addition, COX-2 protein levels and cell migration ability were abolished by ET receptor antagonists. Activation of the mitogen-activated protein kinase (MAPK) and activator protein-1 (AP-1) pathways after ET-1 treatment was demonstrated, and ET-1-induced COX-2 expression and cell migration activity were inhibited by the specific inhibitor and mutant of MAPK and AP-1 cascades. ET-1 increased the binding of c-Jun to the AP-1 element on the COX-2 promoter. Furthermore, knockdown of ET-1 decreased cell metastasis in vitro and in vivo.

Conclusions: Our results indicated that ET-1 enhances the cell migration of chondrosarcoma by increasing COX-2 expression through the ET receptors, MAPK, and AP-1 signal transduction pathway.

General significance: We link high ET-1 and COX-2 expression to chondrosarcoma.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bone Neoplasms / genetics
  • Bone Neoplasms / metabolism*
  • Bone Neoplasms / mortality
  • Cartilage / metabolism
  • Cartilage / pathology
  • Cell Line, Tumor
  • Cell Movement*
  • Chondrosarcoma / genetics
  • Chondrosarcoma / metabolism*
  • Chondrosarcoma / pathology
  • Cyclooxygenase 2 / biosynthesis*
  • Cyclooxygenase 2 / genetics
  • Endothelin-1 / genetics
  • Endothelin-1 / metabolism*
  • Gene Expression Regulation, Enzymologic*
  • Gene Expression Regulation, Neoplastic*
  • Gene Knockdown Techniques
  • Humans
  • MAP Kinase Signaling System / genetics
  • Proto-Oncogene Proteins c-jun / genetics
  • Proto-Oncogene Proteins c-jun / metabolism
  • Transcription Factor AP-1 / genetics
  • Transcription Factor AP-1 / metabolism
  • Up-Regulation / genetics

Substances

  • Endothelin-1
  • Proto-Oncogene Proteins c-jun
  • Transcription Factor AP-1
  • Cyclooxygenase 2
  • PTGS2 protein, human