A novel function of CXCL13 to stimulate RANK ligand expression in oral squamous cell carcinoma cells

Mol Cancer Res. 2009 Aug;7(8):1399-407. doi: 10.1158/1541-7786.MCR-08-0589. Epub 2009 Aug 11.

Abstract

Oral squamous cell carcinomas (OSCC) are malignant tumors with a potent activity of local bone invasion/osteolysis. The chemokine ligand, CXCL13, has been identified as a prognostic marker for OSCC development and progression. Here in, we show that recombinant hCXCL13 treatment of OSCC cells stimulates (5-fold) RANK ligand (RANKL), a critical bone resorbing osteoclastogenic factor expression. Anti-CXCR5 chemokine receptor antibody abrogates CXCL13-induced RANKL expression in these cells. Also, CXCL13 stimulated (3.0-fold) hRANKL gene promoter activity in SCC14a cells. SuperArray screening for transcription factors by real-time RT-PCR identified significant increase in the levels of c-Jun and NFATc3 mRNA expression in CXCL13-stimulated OSCC cells. CXCL13 treatment significantly increased (3.5-fold) phospho-c-Jun levels in these cells and a c-Jun-NH(2)-kinase inhibitor abolished CXCL13-stimulated RANKL expression. Furthermore, we show that CXCL13 stimulation induced nuclear translocation of NFATc3 in OSCC cells. Chromatin-immune precipitation assay confirmed NFATc3 binding to the RANKL promoter region. We also show that overexpression of NFATc3 stimulates RANKL expression/promoter activity and that siRNA suppression of NFATc3 abolished CXCL13-stimulated RANKL expression. Thus, our results suggest that NFATc3 is a downstream target of the CXCL13/CXCR5 axis to stimulate RANKL expression in OSCC cells and implicates CXCL13 as a potential therapeutic target to prevent OSCC bone invasion/osteolysis.

MeSH terms

  • Carcinoma, Squamous Cell / genetics*
  • Cell Line, Tumor
  • Chemokine CXCL13 / metabolism*
  • Chemokine CXCL13 / pharmacology
  • Gene Expression Regulation, Neoplastic / drug effects
  • Humans
  • Mouth Neoplasms / genetics*
  • NFATC Transcription Factors / genetics
  • NFATC Transcription Factors / metabolism
  • Phosphoproteins / metabolism
  • Promoter Regions, Genetic
  • Proto-Oncogene Proteins c-jun / metabolism
  • RANK Ligand / genetics*
  • RANK Ligand / metabolism*
  • Transcription Factors / genetics
  • Transcription Factors / metabolism

Substances

  • CXCL13 protein, human
  • Chemokine CXCL13
  • NFATC Transcription Factors
  • NFATC3 protein, human
  • Phosphoproteins
  • Proto-Oncogene Proteins c-jun
  • RANK Ligand
  • Transcription Factors