克隆法研究辐射诱发人外周血淋巴细胞HPRT基因突变  

STUDY ON THE RADIATION-INDUCED HPRT GENE LOCUS MUTATION IN HUMAN PERIPHERAL BLOOD LYMPHOCYTES BY USING THE COLONING METHOD

作　　者：崔凤梅[1] 赵经涌[1] 劳勤华[1] 王六一[1] 洪承皎[1] 宁萍[1] 

机构地区：[1]苏州大学放射医学与公共卫生学院,215007

出　　处：《辐射防护》2005年第1期36-39,共4页Radiation Protection

基　　金：国防科工委预研项目 (Y5 5 7336 2 )

摘　　要：本文目的是用克隆法研究γ射线诱发人外周血淋巴细胞次黄嘌呤鸟嘌呤磷酸抗糖转移酶(HPRT)基因突变的量效关系。从健康成年人外周血分离淋巴细胞 ,每份分成两等份 ,其中一份加滋养细胞 ,另一份不加滋养细胞 ,种于 96孔板 ,进行淋巴细胞的克隆和突变细胞的筛选 ,比较二者结果 ;剂量效应关系实验为取一健康成年男子外周血 2 5mL分成 5等份 ,分别用 0、1、2、4和 6Gy照射 ,分离淋巴细胞 ,种于 96孔板 ,不加滋养细胞条件下观察淋巴细胞的克隆效率和HPRT基因突变频率的变化。结果表明加滋养细胞组的克隆效率高于不加滋养细胞组 (p <0 0 1 ) ,但二者HPRT基因突变频率无差别 (p >0 0 5 )。在一定剂量范围内 ,人外周血淋巴细胞克隆效率与照射剂量呈负相关 。The aim of this paper is to use cloning method to investigate the radiation-induced HPPT gene locus mutation in human peripheral blood lymphocytes. The lymphocytes separated from peripheral blood of healthy donors were planted into the wells of microplates for cell colony formation in the medium with or without the feeder cells. The HPRT mutation frequency was measured with the ratio of the colony efficiency with the medium containing 6-TG to that without 6-TG. In the radiation experiment, the peripheral blood was exposed to γ-rays at doses 0,1,2, 4 and 6 Gy before lymphocyte separation and then the separated lymphocytes were treated as above in the well without feeder cells. The results showed that the colony efficiency in the case with feeder cells was significantly higher than that without feeder cells (p<0.01), but the HPRT gene mutation frequencies in the two cases had no significant difference (p>0.05). In the radiation dose range used, the colony efficiency of the lymphocytes was negatively correlated to the radiation dose, while the HPRT mutation frequency is positively correlated.

关 键 词：HPRT基因突变 人外周血淋巴细胞 滋养细胞 照射剂量 克隆效率 血分 健康成年人 次黄嘌呤 负相关 转移酶 

分 类 号：R346[医药卫生—基础医学]