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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 ,这可能与其作用机理有关
In order to study the genetic toxicological effects of acrylamide, single cell clonal culture, two-way screening, multiplex PCR amplification and electrophoresis analysis were used to study the mutation rate and molecular mutation spectrum of hprt gene in HL 60 and NB4 cells. A high-dose acrylamide group (70 0 mg·L-1) had a clear hprt gene mutation effect on both cells; acrylamide-induced mutations consisted mainly of point mutations and deletions (40.0% to 66.7. %, 33.3% to 60% ), and spontaneous mutations are almost all point mutations (above 90%). There are no whole gene deletions in both cells; deletion mutations can occur on each of the hprt genes. Exons (except exon 7/8) were more concentrated on the 3′ end of the gene, and most of the induced mutations were point mutations and single exon deletions (93.3%, 86.1%). The situation is similar. The results suggest that acrylamide has a weaker effect of inducing hprt mutations, and the induced mutations are not the same as the molecular maps of spontaneous mutations, which may be related to its mechanism of action