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Immunohistochemical study of nuclear factor-κB activity and interleukin-8 abundance in oesophageal adenocarcinoma; a useful strategy for monitoring these biomarkers
  1. G J S Jenkins1,
  2. J Mikhail1,
  3. A Alhamdani2,
  4. T H Brown2,
  5. S Caplin2,
  6. J M Manson3,
  7. R Bowden4,
  8. N Toffazal4,
  9. A P Griffiths4,
  10. J M Parry1,
  11. J N Baxter2
  1. 1
    Swansea School of Medicine, University of Wales Swansea, Swansea, UK
  2. 2
    Department of Surgery, Morriston Hospital, Swansea, UK
  3. 3
    Department of Surgery, Singleton Hospital, Swansea, UK
  4. 4
    Department of Pathology, Swansea NHS Trust, Swansea, UK
  1. Dr G J S Jenkins, Swansea School of Medicine, University of Wales Swansea, Swansea SA28PP, UK; g.j.jenkins{at}


Aims: To determine if immunohistochemistry (IHC) could be used to monitor nuclear factor-κB (NF-κB) activity in oesophageal adenocarcinoma and pre-malignant (Barrett’s) oesophageal tissues, relative to normal oesophageal mucosa. The pro-inflammatory cytokine interleukin-8 (IL-8), a transcriptional target of NF-κB, was also studied to better understand NF-κB functionality; its RNA and protein levels were assessed in oesophageal tissues.

Methods: IHC was employed using an antibody against the nuclear localisation sequence (NLS) of the p65 subunit as well as an antibody against IL-8. To assess NF-κB function, changes in gene expression of NF-κB controlled genes (IL-8 and I-κB) were also assessed in the histological sequence using real-time PCR. More global expression changes were also studied using membrane arrays.

Results: IHC was effective at monitoring overall NF-κB activity and IL-8 abundance. This method also allowed NF-κB activity and IL-8 abundance to be pinpointed in specific cell types. There were significant increases in nuclear NF-κB activity and IL-8 abundance across the histological series. Gene expression analysis also showed consistent up-regulation of IL-8, confirming the IHC data and showing enhanced transcriptional NF-κB activity. I-κB (another NF-κB target) showed down-regulation in dysplastic and adenocarcinoma tissues. Down-regulation of I-κB gene expression may partly explain increased NF-κB activity.

Conclusion: IHC, using antibodies against the NLS of p65, may be useful in monitoring overall NF-κB activity in oesophageal tissues. As IHC is amenable to high-throughput screening (whereas traditional electrophoretic mobility shift assay methods are not), this may lead to the development of a better screening tool for early cancer risk.

  • immunohistochemistry
  • NF-kB
  • IL-8
  • Barrett’s oesophagus
  • adenocarcinoma
  • biomarker

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  • Funding: This work was partly funded by the Welsh Office for Research and Development (Grant WS01/1/009).

  • Competing interests: None.

  • Abbreviations:
    nuclear factor-κB
    nuclear localisation sequence