丙烯酰胺诱导人白血病HL-60和NB_4细胞hprt基因的分子突变谱  

Molecular spectra of acrylamide-induced mutation at hprt locus in human promyelocytic leukemia HL-60 and NB_4 cell lines

作　　者：刘胜学[1] 曹佳[1] 杨梦甦[2] 方志俊[2] 安辉[1] 

机构地区：[1]第三军医大学分子毒理学实验室,重庆400038 [2]香港城市大学生物化学系

出　　处：《中国药理学与毒理学杂志》2001年第4期276-281,共6页Chinese Journal of Pharmacology and Toxicology

基　　金：国家自然科学基金项目 (39970 6 5 0 );军队基金项目 (98M0 90 );教育部优秀青年教师资助计划项目 (2 0 0 0 81);香港城市大学RCP资助项目 (936 0 0 17)&&

摘　　要：为了研究丙烯酰胺的遗传毒理作用 ,采用单细胞克隆培养 ,双向筛选计数 ,多重PCR扩增与电泳分析 ,研究了诱导HL 6 0和NB4 两种细胞hprt基因突变率及分子突变谱 .发现只有丙烯酰胺高剂量组 (70 0mg·L- 1)才对两种细胞有明确的致hprt基因突变作用 ;丙烯酰胺诱发突变主要由点突变和缺失两部分组成 (40 .0 %～ 6 6 .7% ,33.3%～ 6 0 .0 % ) ,而自发突变几乎全是点突变 (90 .0 %以上 ) ,两种细胞均无全基因缺失型 ;缺失突变可以发生于hprt基因上的每个外显子 (除外显子 7/ 8以外 ) ,较集中于基因的 3′末端 ,且诱发突变中绝大多数是点突变与单个外显子缺失 (93.3% ,86 .1% ) ,两种细胞情况类似 .结果提示 ,丙烯酰胺具有较弱的诱导hprt基因突变的作用 ,且诱发突变与自发突变的分子图谱不一样 。The genotoxicity of acrylamide was investigated by methods of single cell clone culturing, two way screening count, multiplex PCR amplification and electrophoresis technique. Acryl amide only showed clear mutagenesis until dose 700 mg·L -1 in HL 60 cells. The most frequent spontaneous mutation was point mutation(≥90.0%) and acrylamide induced mutation mainly included partial deletion and point mutation(respectively 40.0%-66.7%, 33.3%-60.0%). Total gene deletion was not discovered in both of cells. There was deletion mutation in all exons of hprt gene(except 7/8 exon), and toward the 3′ end of the hprt gene. The most frequent acrylamide induced mutations were point mutation and single exon deletion(93.3%, 86.1%). There was no clear difference in both of cells. The results suggest that the spectra of spontaneous and acrylamide induced mutants are different, and the smaller changes in genetic structure have something to do with mechanism.

关 键 词：HPRT基因 丙烯酰胺 诱变 NB4细胞 白血病 HL-60细胞 

分 类 号：R733.7[医药卫生—肿瘤]