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Ribozyme minigene-mediated RAD51 down-regulation increases radiosensitivity of human prostate cancer cells

Ribozyme minigene-mediated RAD51 down-regulation increases radiosensitivity of human prostate cancer cells

Collis, S.J., Tighe, A., Scott, S.D., Roberts, S.A., Hendry, J.H. and Margison, G.P. (2001) Ribozyme minigene-mediated RAD51 down-regulation increases radiosensitivity of human prostate cancer cells. Nucleic Acids Research, 29 (7). pp. 1534-1538. ISSN 0305-1048 (Print), 1362-4962 (Online) (doi:10.1093/nar/29.7.1534)

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Abstract

The strand transferase RAD51 is a component of the homologous recombination repair pathway. To examine the contribution of RAD51 to the genotoxic effects of ionising radiation, we have used a novel ribozyme strategy. A reporter gene vector was constructed so that expression of an inserted synthetic double-stranded ribozyme-encoding oligonucleotide would be under the control of the cytomegalovirus immediate-early gene enhancer/promoter system. The prostate tumour cell line LNCaP was transfected with this vector or a control vector, and a neomycin resistance gene on the vector was used to create geneticin-resistant stable cell lines. Three stable cell lines were shown by western blot analysis to have significant down-regulation of RAD51 to 20–50% of the levels expressed in control cell lines. All three cell lines had a similar increased sensitivity to γ-irradiation by 70 and 40%, respectively, compared to normal and empty vector-transfected cells, corresponding to dose-modifying factors of ∼2.0 and 1.5 in the mid-range of the dose-response curves. The amount of RAD51 protein in transfected cell lines was shown to strongly correlate with the α parameter obtained from fitted survival curves. These results highlight the importance of RAD51 in cellular responses to radiation and are the first to indicate the potential use of RAD51-targeted ribozyme minigenes in tumour radiosensitisation.

Item Type: Article
Uncontrolled Keywords: gene vector, tumour, prostate, RAD51, radiation
Subjects: R Medicine > RC Internal medicine > RC0254 Neoplasms. Tumors. Oncology (including Cancer)
Faculty / Department / Research Group: Faculty of Engineering & Science > Medway School of Pharmacy
Faculty of Engineering & Science
Related URLs:
Last Modified: 14 Jul 2015 16:22
Selected for GREAT 2016: None
Selected for GREAT 2017: None
Selected for GREAT 2018: None
URI: http://gala.gre.ac.uk/id/eprint/8474

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