用多重PCR分析^(60)Coγ射线引起人白血病细胞HPRT基因突变  

Molecular analysis of ^(60 )Co-gamma ray |induced mutation at HPRT locus in human promyelocytic leukemia cells\

作　　者：刘胜学[1] 曹佳[1] 安辉[1] 周紫恒 孙华明[1] 

机构地区：[1]第三军医大学预防医学系分子毒理室,重庆400038

出　　处：《中华放射医学与防护杂志》2000年第4期225-227,共3页Chinese Journal of Radiological Medicine and Protection

基　　金：国家自然科学基金!资助项目 (39970 6 5 0 )

摘　　要：目的　探讨60 Coγ射线诱导人HPRT基因突变的分子图谱和发生机理 ,以及与辐射抗肿瘤的关系。方法　采用单细胞克隆培养、双向筛选计数、多重PCR扩增和电泳分析等方法。结果①随着辐射剂量的增加 ,细胞接种存活率下降 ,突变频率升高。②自发突变中 92 3%是点突变 ,而γ射线诱发突变主要由缺失与点突变两部分组成 (两者分别为 6 1 7%和 38 3 % )。③缺失突变可以发生于HPRT基因的每个外显子 ,但诱发突变中绝大多数是多个外显子的连锁缺失 (97 9% )。结论γ射线诱发HPRT基因突变图谱与自发突变不一样 。Objective\ To explore the spectra and mechanism of human HPRT gene mutation induced by 60 Co γ\|rays and its relation with anti\|tumor effect of radiation. Methods\ Single cell clone culturing,two\|way screening count,multiplex PCR amplification and electrophoresis technique were used. Results\ (1)When doses were increasing,cell plating efficiency reduced and mutation frequency increased.(2)The most frequent spontaneous mutations were point mutation (92 3%) and gamma\|ray\|induced mutation,including mainly partial deletion and point mutation(61\^7% and 38\^3%,respectively).(3)There were deletion mutations in all 9 exons of HPRT gene and the most of gamma\|ray\|induced mutations were chain deletion with multiplex exons (97\^9%). Conclusion\ The spectra of spontaneous and gamma\|ray\|induced mutants were different.The bigger changes in genetic structure are related to the anti\|tumor mechanism of radiation.\;

关 键 词：白血病 HL-60细胞 HPRT基因 多重PCR 钴60γ射线 

分 类 号：R730.3[医药卫生—肿瘤]