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

2.50
Hdl Handle:
http://hdl.handle.net/10541/85548
Title:
Ribozyme minigene-mediated RAD51 down-regulation increases radiosensitivity of human prostate cancer cells.
Authors:
Collis, S J; Tighe, A; Scott, S D; Roberts, Stephen A; Hendry, Jolyon H; Margison, Geoffrey P
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 gamma-irradiation by 70 and 40%, respectively, compared to normal and empty vector-transfected cells, corresponding to dose-modifying factors of approximately 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 alpha 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.
Affiliation:
Carcinogenesis Group and Experimental Radiation Oncology Group, Cancer Research Campaign, Paterson Institute for Cancer Research, Wilmslow Road, Manchester M20 4BX, UK.
Citation:
Ribozyme minigene-mediated RAD51 down-regulation increases radiosensitivity of human prostate cancer cells. 2001, 29 (7):1534-8 Nucleic Acids Res.
Journal:
Nucleic Acids Research
Issue Date:
1-Apr-2001
URI:
http://hdl.handle.net/10541/85548
PubMed ID:
11266555
Type:
Article
Language:
en
ISSN:
1362-4962
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorCollis, S Jen
dc.contributor.authorTighe, Aen
dc.contributor.authorScott, S Den
dc.contributor.authorRoberts, Stephen Aen
dc.contributor.authorHendry, Jolyon Hen
dc.contributor.authorMargison, Geoffrey Pen
dc.date.accessioned2009-11-06T14:42:02Z-
dc.date.available2009-11-06T14:42:02Z-
dc.date.issued2001-04-01-
dc.identifier.citationRibozyme minigene-mediated RAD51 down-regulation increases radiosensitivity of human prostate cancer cells. 2001, 29 (7):1534-8 Nucleic Acids Res.en
dc.identifier.issn1362-4962-
dc.identifier.pmid11266555-
dc.identifier.urihttp://hdl.handle.net/10541/85548-
dc.description.abstractThe 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 gamma-irradiation by 70 and 40%, respectively, compared to normal and empty vector-transfected cells, corresponding to dose-modifying factors of approximately 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 alpha 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.en
dc.language.isoenen
dc.subjectProstatic Canceren
dc.subjectCultured Tumour Cellsen
dc.subject.meshBase Sequence-
dc.subject.meshCell Division-
dc.subject.meshCytomegalovirus-
dc.subject.meshDNA Repair-
dc.subject.meshDNA-Binding Proteins-
dc.subject.meshDose-Response Relationship, Radiation-
dc.subject.meshDown-Regulation-
dc.subject.meshGene Expression Regulation, Neoplastic-
dc.subject.meshGreen Fluorescent Proteins-
dc.subject.meshHumans-
dc.subject.meshImmediate-Early Proteins-
dc.subject.meshLuminescent Proteins-
dc.subject.meshMale-
dc.subject.meshPromoter Regions, Genetic-
dc.subject.meshProstatic Neoplasms-
dc.subject.meshRNA, Catalytic-
dc.subject.meshRNA, Messenger-
dc.subject.meshRad51 Recombinase-
dc.subject.meshRecombinant Fusion Proteins-
dc.subject.meshRecombination, Genetic-
dc.subject.meshTumor Cells, Cultured-
dc.titleRibozyme minigene-mediated RAD51 down-regulation increases radiosensitivity of human prostate cancer cells.en
dc.typeArticleen
dc.contributor.departmentCarcinogenesis Group and Experimental Radiation Oncology Group, Cancer Research Campaign, Paterson Institute for Cancer Research, Wilmslow Road, Manchester M20 4BX, UK.en
dc.identifier.journalNucleic Acids Researchen
All Items in Christie are protected by copyright, with all rights reserved, unless otherwise indicated.