细胞照射后可产生DNA链断裂、DNA-DNA交联、DNA-蛋白质交联等重要的DNA结构损伤,最终可导致DNA高级结构-DNA超螺旋结构状态的改变,而引发DNA复制、表达等一系列改变.参考国外报导,建立了单细胞电泳法(single cell gel electrophoresis assay),并辅以图象分析技术,可快速检测低达0.1Gy剂量所致DNA结构损伤,并得到了较好的剂量-效应关系,可望成为生物剂量计,用于环境低剂量辐射的监测.
Irradiated cells express DNA structural damages. such as DNA double-strand break. DNA-DNA crosslink and DNA-protein crosslink etc. These damages result to changes of DNA supercoil state, which trigger a series changes of DNA replication and expression. Many methods to test DNA damages have been established, roughly can be devided into two kinds according to the DNA materials used. The first use the naked DNA extracted from cells and free from DNA-binding materials existed in cell. The second one uses nucleoid for research, here the detergents and hypertonic salt buffers were used to remove nuclear envelope and a part of nuclear proteins, nuclear DNA remains appropriate tangled loop and binds to residual nucle-arskeleton. This DNA structure is beneficial to research damage effects on DNA structure, single cell gel electrophoresis belongs to the latter. It also named comet assay because its cell electrophoresis shape looks like a comet. It can test not only DNA stand break but also measure DNA structure changes resulted from stand bleak. According to oversea reports, with slight modification, single cell gel electrophoresis assay has been established Employing image analysis system. fast quntitative measurement of DNA structure changes of single cell irradiated as low as 0.1Gy can be given with a well correlated dose-respones relationship. After further study, the method might be developed as a kind of biodosimeter for application of monitoring enviromental low level irradiation.
罗瑛,孙志贤,杨瑞彪,张振声.辐射后单个细胞DNA结构变化的定量检测[J].生物化学与生物物理进展,1994,21(5):451-453
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