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

AP McGlynn, GR Wasson, S O'Reilly, Valerie McKelvey-Martin, Helene McNulty, JJ Strain, George McKerr, F Mullan, N Mahmud, J Scott, DG Weir, Stephen Downes

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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.
LanguageEnglish
Pages895-901
JournalBRITISH JOURNAL OF CANCER
Volume88
Issue number6
DOIs
Publication statusPublished - Mar 2003

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Comet Assay
Biopsy
Endoscopy
Colon
Single-Stranded DNA Breaks
Neoplasms
DNA
Alkalies
Bromodeoxyuridine
Cultured Cells
Carcinogenesis
Biomarkers
Diet

Cite this

McGlynn, AP ; Wasson, GR ; O'Reilly, S ; McKelvey-Martin, Valerie ; McNulty, Helene ; Strain, JJ ; McKerr, George ; Mullan, F ; Mahmud, N ; Scott, J ; Weir, DG ; Downes, Stephen. / Detection of replicative integrity in small colonic biopsies using the BrdUrd comet assay. In: BRITISH JOURNAL OF CANCER. 2003 ; Vol. 88, No. 6. pp. 895-901.
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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.",
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McGlynn, AP, Wasson, GR, O'Reilly, S, McKelvey-Martin, V, McNulty, H, Strain, JJ, McKerr, G, Mullan, F, Mahmud, N, Scott, J, Weir, DG & Downes, S 2003, 'Detection of replicative integrity in small colonic biopsies using the BrdUrd comet assay', BRITISH JOURNAL OF CANCER, vol. 88, no. 6, pp. 895-901. https://doi.org/10.1038/sj.bjc.6600836

Detection of replicative integrity in small colonic biopsies using the BrdUrd comet assay. / McGlynn, AP; Wasson, GR; O'Reilly, S; McKelvey-Martin, Valerie; McNulty, Helene; Strain, JJ; McKerr, George; Mullan, F; Mahmud, N; Scott, J; Weir, DG; Downes, Stephen.

In: BRITISH JOURNAL OF CANCER, Vol. 88, No. 6, 03.2003, p. 895-901.

Research output: Contribution to journalArticle

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T1 - Detection of replicative integrity in small colonic biopsies using the BrdUrd comet assay

AU - McGlynn, AP

AU - Wasson, GR

AU - O'Reilly, S

AU - McKelvey-Martin, Valerie

AU - McNulty, Helene

AU - Strain, JJ

AU - McKerr, George

AU - Mullan, F

AU - Mahmud, N

AU - Scott, J

AU - Weir, DG

AU - Downes, Stephen

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AB - 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.

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