Development of a novel rapid assay to assess the fidelity of DNA double-strand-break repair in human tumour cells.
AuthorsCollis, S J
Sangar, Vijay K
Roberts, Stephen A
Clarke, Noel W
Hendry, Jolyon H
Margison, Geoffrey P
AffiliationCRC Experimental Radiation Oncology Group, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Wilmslow Road, Manchester M20 4BX, UK.
MetadataShow full item record
AbstractCellular survival following ionising radiation-mediated damage is primarily a function of the ability to successfully detect and repair DNA double-strand breaks (DSBs). Previous studies have demonstrated that radiosensitivity, determined as a reduction in colony forming ability in vitro, may be related to the incorrect repair (misrepair) of DSBs. The novel rapid dual fluorescence (RDF) assay is a plasmid-based reporter system that rapidly assesses the correct rejoining of a restriction-enzyme produced DSBs within transfected cells. We have utilised this novel assay to determine the fidelity of DSB repair in the prostate tumour cell line LNCaP, the bladder tumour cell line MGH-U1 and a radiosensitive subclone S40b. The two bladder cell lines have been shown in previous studies to differ in their ability to correctly repair plasmids containing a single DSB. Using the RDF assay we found that a substantial portion of LNCaP cells [80.4 +/- 5.3(standard error)%] failed to reconstitute reporter gene expression; however, there was little difference in this measure of DSB repair fidelity between the two bladder cell lines (48.3 +/- 3.5% for MGH-U1; 39.9 +/- 8.2% for S40b). The RDF assay has potential to be developed to study the relationship between DSB repair fidelity and radiosensitivity as well as the mechanisms associated with this type of repair defect.
CitationDevelopment of a novel rapid assay to assess the fidelity of DNA double-strand-break repair in human tumour cells. 2002, 30 (2):E1 Nucleic Acids Res.
JournalNucleic Acids Research
- The repair fidelity of restriction enzyme-induced double strand breaks in plasmid DNA correlates with radioresistance in human tumor cell lines.
- Authors: Powell SN, McMillan TJ
- Issue date: 1994 Jul 30
- Differential level of DSB repair fidelity effected by nuclear protein extracts derived from radiosensitive and radioresistant human tumour cells.
- Authors: Britten RA, Liu D, Kuny S, Allalunis-Turner MJ
- Issue date: 1997
- Rejoining of DNA double-strand breaks after the introduction of chromosome 11 into a radiosensitive bladder carcinoma cell line.
- Authors: Hofseth LJ, Tsang SS, Rosin MP
- Issue date: 1997 Jan 31
- Radiation-induced DNA double-strand break rejoining in human tumour cells.
- Authors: Núñez MI, Villalobos M, Olea N, Valenzuela MT, Pedraza V, McMillan TJ, Ruiz de Almodóvar JM
- Issue date: 1995 Feb
- A DNA repair defect in a radiation-sensitive clone of a human bladder carcinoma cell line.
- Authors: Powell SN, Whitaker SJ, Edwards SM, McMillan TJ
- Issue date: 1992 Jun