A novel nuclear factor-kappaB gene signature is differentially expressed in head and neck squamous cell carcinomas in association with TP53 status

Clin Cancer Res. 2007 Oct 1;13(19):5680-91. doi: 10.1158/1078-0432.CCR-07-0670.


Purpose: To determine if gene signatures differentially expressed in head and neck squamous cell carcinomas (HNSCC) are related to alterations in transcription factors nuclear factor-kappaB (NF-kappaB) and TP53 previously associated with decreased cell death, response to therapy, and worse prognosis.

Experimental design: Unique gene signatures expressed by HNSCC lines were identified by cDNA microarray, principal components, and cluster analyses and validated by quantitative reverse transcription-PCR (RT-PCR) and in situ hybridization. Bioinformatic analysis of the promoters and ontogeny of these clustered genes was done. Expression of proteins encoded by genes of a putative NF-kappaB signature, NF-kappaB p65, and TP53 were examined in HNSCC tissue specimens by immunostaining. Predicted promoter binding and modulation of expression of candidate NF-kappaB genes and cell survival were evaluated by p65 chromatin immunoprecipitation (ChIP) and small interfering RNA (siRNA) knockdown.

Results: Two groups of HNSCC exhibiting distinct gene signatures were identified: cluster A enriched for histone genes, with a higher prevalence of TP53 promoter binding motifs; and cluster B enriched for injury response genes with NF-kappaB regulatory motifs. Coexpression of cluster B proteins was observed with strong NF-kappaB phospho-p65 and weak TP53 staining, and NF-kappaB phospho-p65 was inversely associated with TP53 (P = 0.02). Promoter binding of the NF-kappaB signature genes was confirmed by p65 ChIP, and down-modulation of their expression and cell death were induced by p65 siRNA.

Conclusion: NF-kappaB promotes expression of a novel NF-kappaB-related gene signature and cell survival in HNSCC that weakly express TP53, a subset previously associated with inactivated wild-type TP53, greater resistance to chemoradiotherapy, and worse prognosis.

Publication types

  • Research Support, N.I.H., Intramural

MeSH terms

  • Carcinoma, Squamous Cell / genetics*
  • Cell Line, Tumor
  • Computational Biology
  • Gene Expression Profiling
  • Gene Expression Regulation, Neoplastic*
  • Genes, p53 / genetics*
  • Head and Neck Neoplasms / genetics*
  • Humans
  • NF-kappa B / metabolism*
  • Oligonucleotide Array Sequence Analysis
  • Prognosis
  • RNA, Small Interfering / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Treatment Outcome
  • Tumor Suppressor Protein p53 / genetics*


  • NF-kappa B
  • RNA, Small Interfering
  • TP53 protein, human
  • Tumor Suppressor Protein p53