Transfer of Ku86 RNA antisense decreases the radioresistance of human fibroblasts
| Autor: | Muriel Le Romancer, Jacques Bernier, Nicolas Foray, Elisabetta Marangoni, Sabine Vaganay, Michel Barrois, Patrick Calsou, Catherine Muller, Bassam Abdulkarim, Bernard Salles, Jean Bourhis, Sétha Douc-Rasy |
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| Rok vydání: | 2000 |
| Předmět: |
Cancer Research
DNA Repair Cell Survival Genetic enhancement Population Blotting Western Molecular Sequence Data DNA-Activated Protein Kinase Protein Serine-Threonine Kinases Radiation Tolerance Cell Line chemistry.chemical_compound Radioresistance Complementary DNA Humans RNA Antisense Radiosensitivity Amino Acid Sequence Kinase activity education Molecular Biology Ku Autoantigen education.field_of_study Micronucleus Tests Chemistry fungi DNA Helicases Gene Transfer Techniques Nuclear Proteins Antigens Nuclear Transfection Fibroblasts Molecular biology Clone Cells DNA-Binding Proteins Kinetics Cesium Radioisotopes Gamma Rays Molecular Medicine DNA |
| Zdroj: | Cancer gene therapy. 7(2) |
| ISSN: | 0929-1903 |
| Popis: | Ku86 has been shown to be involved in DNA double-strand break (DSB) repair and radiosensitivity in rodents, but its role in human cells is still under investigation. The purpose of this study was to evaluate the radiosensitivity and DSB repair after transfection of a Ku86-antisense in a human fibroblast cell line. Simian virus 40-transformed MRC5V1 human fibroblasts were transfected with a vector (pcDNA3) containing a Ku86-antisense cDNA. The main endpoints were Ku86 protein level, Ku DNA end-binding and DNA protein kinase activity, clonogenic survival, and DSB repair kinetics. After transfection of the Ku86-antisense, decreased Ku86 protein expression, Ku DNA end-binding activity, and DNA protein kinase activity were observed in the uncloned cellular population. The fibroblasts transfected with the Ku86-antisense showed also a radiosensitive phenotype, with a surviving fraction at 2 Gy of 0.29 compared with 0.75 for the control and 20% of unrepaired DSB observed at 24 hours after irradiation compared with 0% for the control. Several clones were also isolated with a decreased level of Ku86 protein, a surviving fraction at 2 Gy between 0.05 and 0.40, and 10–20% of unrepaired DSB at 24 hours. This study is the first to show the implication of Ku86 in DSB repair and in the radiosensitivity of human cells. This investigation strongly suggests that Ku86 could constitute an appealing target for combining gene therapy and radiation therapy. |
| Databáze: | OpenAIRE |
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