NF-kappaB p50 promotes tumor cell invasion through negative regulation of invasion suppressor gene CRMP-1 in human lung adenocarcinoma cells

Biochem Biophys Res Commun. 2008 Nov 14;376(2):283-7. doi: 10.1016/j.bbrc.2008.08.144. Epub 2008 Sep 7.

Abstract

Lung adenocarcinoma Cl1-5 cells were selected from parental Cl1-0 cells based on their high metastatic potential. In a previous study, CRMP-1, an invasion suppressor gene, was shown to be suppressed in Cl1-5 cells. However, the regulation of CRMP-1 expression has not been explored. In this study, we showed nuclear factor-kappaB controls CRMP-1 expression. The electromobility shift assay showed that while Cl1-0 cells exhibited low NF-kappaB activity in response to TNF-alpha, an abundance of basal and TNF-alpha-induced NF-kappaB-DNA complex was detected in Cl1-5 cells. Supershift-coupled EMSA and Western blotting of nuclear proteins, however, revealed p50 protein, but not classic p65/p50 heterodimer in the complex. ChIP and EMSA demonstrated that p50 binds to a kappaB site residing between -1753 and -1743 of the CRMP-1 promoter region. Transfection of antisense p50 gene into Cl1-5 cells increased the CRMP-1 protein level and decreased the invasive activity of Cl1-5 cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenocarcinoma / genetics
  • Adenocarcinoma / pathology*
  • Cell Line, Tumor
  • Electrophoretic Mobility Shift Assay
  • Gene Expression Regulation, Neoplastic*
  • Genes, Tumor Suppressor*
  • Humans
  • Lung Neoplasms / genetics
  • Lung Neoplasms / pathology*
  • NF-kappa B p50 Subunit / metabolism*
  • Neoplasm Invasiveness
  • Nerve Tissue Proteins / genetics*
  • Promoter Regions, Genetic
  • Transcription Factor RelA / metabolism
  • Tumor Necrosis Factor-alpha / pharmacology

Substances

  • CRMP1 protein, human
  • NF-kappa B p50 Subunit
  • Nerve Tissue Proteins
  • Transcription Factor RelA
  • Tumor Necrosis Factor-alpha