Prolongation of carrageenan-induced inflammation in human colonic epithelial cells by activation of an NFκB-BCL10 loop

Biochim Biophys Acta. 2012 Aug;1822(8):1300-7. doi: 10.1016/j.bbadis.2012.05.001. Epub 2012 May 8.

Abstract

Carrageenan, a sulfated polysaccharide that is widely used as a food additive, induces inflammatory responses in animal models and human cells. The carrageenan-induced inflammatory cascades involve toll-like receptor (TLR)4- and B-cell leukemia/lymphoma (BCL)10-dependent activation of NF-κB, leading to increased IL-8 production. Translocations involving BCL10 in the mucosa-associated lymphoid tissue (MALT) lymphomas are associated with constitutive activation of NF-κB. This report presents a mechanism by which carrageenan exposure leads to prolonged activation of both BCL10 and NF-κB in human colonic epithelial cells. Study findings demonstrate that nuclear RelA and RelB bind to an NF-κB binding motif in the BCL10 promoter in human colonic epithelial NCM460 and HT-29 cells. In vitro oligonucleotide binding assay, non-radioactive gel shift assay, and chromatin immunoprecipitation (ChIP) indicate binding of RelA and RelB to the BCL10 promoter. Prolonged inflammation follows activation of the BCL10-NFκB inflammatory loop in response to carrageenan, shown by increased BCL10, RelA, and IL-8 for 36 to 48h and increased RelB for 24h following withdrawal of carrageenan after 12h. In contrast, exposure to dextran sulfate sodium, which does not cause inflammation through TLR4 and BCL10 in the colonic epithelial cells, did not provoke prolonged activation of inflammation. The carrageenan-enhanced BCL10 promoter activity was blocked by caffeic acid phenethyl ester (CAPE) and MB-132 which inhibit NF-κB activation. These results indicate that NF-κB binding to the BCL10 promoter can lead to prolonged activation of the carrageenan-induced inflammatory cascade by a transcriptional mechanism involving an NF-κB-BCL10 loop.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Adaptor Proteins, Signal Transducing / genetics
  • Adaptor Proteins, Signal Transducing / metabolism*
  • B-Cell CLL-Lymphoma 10 Protein
  • Carrageenan / pharmacology*
  • Cell Line
  • Colitis / chemically induced
  • Colitis / genetics
  • Colitis / metabolism*
  • Colon / drug effects*
  • Colon / metabolism*
  • Colon / pathology
  • Epithelial Cells / drug effects
  • Epithelial Cells / metabolism
  • HT29 Cells
  • Humans
  • NF-kappa B / metabolism*
  • Phosphorylation
  • Promoter Regions, Genetic
  • Signal Transduction
  • Transcription Factor RelA / metabolism
  • Transcription Factor RelB / metabolism
  • Transfection

Substances

  • Adaptor Proteins, Signal Transducing
  • B-Cell CLL-Lymphoma 10 Protein
  • BCL10 protein, human
  • NF-kappa B
  • RELA protein, human
  • RELB protein, human
  • Transcription Factor RelA
  • Transcription Factor RelB
  • Carrageenan