Identification of NF-κB-regulated genes induced by TNFα utilizing expression profiling and RNA interference

Anwu Zhou, Shane Scoggin, Richard B. Gaynor, Noelle Sevilir Williams

Research output: Contribution to journalArticle

147 Scopus citations

Abstract

Tumor necrosis factor α (TNFα) is a proinflammatory cytokine with important roles in regulating inflammatory responses as well as cell cycle proliferation and apoptosis. Although TNFα stimulates apoptosis, it also activates the transcription factor NF-κB, and studies have shown that inhibition of NF-κB potentiates the cytotoxicity of TNFα. Since several chemotherapy agents act like TNFα to both promote apoptosis and activate NF-κB, understanding the role of NF-κB in suppressing apoptosis may have significant clinical applications. To understand the effects of stimulation with TNFα and the role of NK-κB in regulating this response, a 23k human cDNA microarray was used to screen TNFα-inducible genes in HeLa cells. Real-time PCR verified expression changes in 16 of these genes and revealed three distinct temporal patterns of expression after TNFα stimulation. Using RNA interference to disrupt expression of the p65 subunit of NF-κB, all but two of the genes were shown to depend on this transcription factor for their expression, which correlated well with the existence of NF-κB binding sites in most of their promoters. Inflammatory, proapoptotic, and antiapoptotic genes were all shown to be regulated by NF-κB, demonstrating the wide variety of targets activated by NF-κB signaling and the necessity of differentiating among these genes for therapeutic purposes.

Original languageEnglish (US)
Pages (from-to)2054-2064
Number of pages11
JournalOncogene
Volume22
Issue number13
DOIs
StatePublished - Apr 3 2003

Keywords

  • Apoptosis
  • Gene expression profiles
  • NF-κB
  • RNA interference
  • TNFα
  • cDNA microarrays

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cancer Research

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