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Immunohistochemical study of nuclear factor- B activity and interleukin-8 abundance in oesophageal adenocarcinoma; a useful strategy for monitoring these biomarkers

Jenkins, G J S, Mikhail, J, Alhamdani, A, Brown, T H, Caplin, S, Manson, J M, Bowden, R, Toffazal, N, Griffiths, A P, Parry, J M and Baxter, J N 2007. Immunohistochemical study of nuclear factor- B activity and interleukin-8 abundance in oesophageal adenocarcinoma; a useful strategy for monitoring these biomarkers. Journal of Clinical Pathology 60 (11) , pp. 1232-1237. 10.1136/jcp.2006.043976

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Abstract

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.

Item Type: Article
Date Type: Published Online
Status: Published
Schools: Biosciences
Publisher: BMJ Publishing Group
ISSN: 0021-9746
Date of Acceptance: 8 December 2006
Last Modified: 08 Nov 2018 15:15
URI: https://orca.cardiff.ac.uk/id/eprint/116518

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