Mlejnek P, Kozubek S
Institute of Biophysics AS CR, Brno, Czech Republic.
Radiat Environ Biophys. 1996 Aug;35(3):171-7. doi: 10.1007/s004110050027.
Fluorometric analysis of DNA unwinding (FADU)-a sensitive technique for the detection of strand breaks in DNA-has been modified and used for the detailed investigation of repair kinetics of DNA-strand breaks arising under different conditions in Ehrlich ascites tumour (EAT) cells irradiated by gamma-rays or ultraviolet (UV) radiation. The repair kinetics of DNA-strand breaks induced in EAT cells by gamma-radiation was measured at radiation doses of 8, 20 and 50 Gy. We found complex repair curves in all cases, probably reflecting the combined processes of break rejoining and break generation during repair. In order to affect the above-mentioned processes, we have used different conditions of repair and different types of radiation. Lowering of the temperature of incubation and treating the cells by 5-fluoro-2'-deoxyuridine (FUdR) lead to complex changes of the repair curve with a reduced "wave" pattern. In order to change the type of damage to DNA, we used UV radiation (254 nm, 10 and 20 J/m2). Detailed studies of the repair kinetics showed that the repair curve for 10 J/m2 had a second maximum within 70 min after irradiation.
DNA解旋荧光分析(FADU)——一种检测DNA链断裂的灵敏技术——已被改进,并用于详细研究在不同条件下受γ射线或紫外线(UV)辐射的艾氏腹水瘤(EAT)细胞中DNA链断裂的修复动力学。在8、20和50 Gy的辐射剂量下测量了γ辐射诱导EAT细胞中DNA链断裂的修复动力学。我们在所有情况下都发现了复杂的修复曲线,这可能反映了修复过程中断裂重新连接和断裂产生的联合过程。为了影响上述过程,我们采用了不同的修复条件和不同类型的辐射。降低孵育温度并用5-氟-2'-脱氧尿苷(FUdR)处理细胞会导致修复曲线发生复杂变化,“波”模式减少。为了改变对DNA的损伤类型,我们使用了紫外线辐射(254 nm,10和20 J/m²)。对修复动力学的详细研究表明,10 J/m²的修复曲线在照射后70分钟内有第二个最大值。