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Detection of replicative integrity in small colonic biopsies using the BrdUrd comet assay

Biomedical Sciences Research Institute Computer Science Research Institute Environmental Sciences Research Institute Nanotechnology & Advanced Materials Research Institute

McGlynn, AP, Wasson, GR, O'Reilly, S, McKelvey-Martin, Valerie, McNulty, Helene, Strain, JJ, McKerr, George, Mullan, F, Mahmud, N, Scott, J, Weir, DG and Downes, Stephen (2003) Detection of replicative integrity in small colonic biopsies using the BrdUrd comet assay. BRITISH JOURNAL OF CANCER, 88 (6). pp. 895-901. [Journal article]

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DOI: 10.1038/sj.bjc.6600836

Abstract

The alkaline single-cell gel electrophoresis or comet assay is a relatively simple method of measuring DNA single-strand breaks and alkali-labile sites in individual cells. Previously, we have used a combination of this with bromodeoxyuridine labelling of DNA and immunolocalisation of the BrdUrd to show that DNA replicative integrity can be assessed in single cultured cells. This study demonstrates the application of the technique to single cells derived from small human colonic biopsies isolated at routine endoscopy. A high level of reproducibility within replicate comet slides and between comet slides prepared from various colonic sites within a single patient is shown. Preliminary results demonstrate that defects in replication can be detected in tumour and premalignant colonic tissue adjacent to the tumour, suggesting that alterations in replicative integrity are an early event in neoplasia, appearing in premalignant mucosal cells. This development deems the BrdUrd comet assay suitable as an ex vivo molecular end point that can be measured easily in tissue collected by biopsy at routine colonic endoscopy. Thus, the BrdUrd comet assay has the potential to facilitate trial investigations of diet- or environment-related factors that may affect replicative integrity in the colon and provides a novel biomarker for colon carcinogenesis. (C) 2003 Cancer Research UK.

Item Type:Journal article
Faculties and Schools:Faculty of Life and Health Sciences
Faculty of Life and Health Sciences > School of Biomedical Sciences
Faculty of Life and Health Sciences > School of Pharmacy and Pharmaceutical Science
Research Institutes and Groups:Biomedical Sciences Research Institute
Biomedical Sciences Research Institute > Molecular Medicine
Biomedical Sciences Research Institute > Molecular Medicine > Nano Systems Biology
ID Code:3393
Deposited By:Professor Stephen Downes
Deposited On:15 Dec 2009 16:32
Last Modified:04 Dec 2012 12:23

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