卷烟烟气及其主要有害成分诱发细胞基因突变的研究

HPRT Gene Locus Mutation of HPV-18 Immortalized Human Bronchial Epithelial Cells Induced by Cigarette Smoke Condense and Its Major Harmful Component

目的研究卷烟烟气凝集物(CSC)及其主要有害成分吡咯叮酮基亚硝胺(NNK)诱发细胞基因突变的作用,为降低卷烟危害技术研究提供有益线索。方法以永生化的人支气管上皮细胞(BEP2D)为靶细胞;某种知名品牌卷烟(混合型,标示焦油量为8mg/支)用于制备CSC,并通过气相色谱-热能分析联用仪(GC-TEA)检测其NNK含量;用多核细胞法检测细胞次黄嘌呤鸟嘌呤磷酸核糖转移酶(HPRT)基因突变率(mutationfrequencyMF)。结果该卷烟制备的CSC测定量为(7.93±0.89)mg/支,与标示焦油量相当,其中NNK含量为(0.046±0.008)μg/支;NNK和CSC的浓度(x1和x2,μg/ml)诱发BEP2D细胞HPRT基因突变率(y1和y2,‰)的剂量-效应关系分别为:y1=0.016x1+1.012和y2=0.0061x2+1.042,CSC中NNK诱发细胞HPRT基因突变率的作用仅占0.002%。结论卷烟烟气可诱发细胞HPRT基因突变,但其中NNK的作用不是主要的。

Objective To study the HPRT gene locus mutations induced by cigarette smoke and its major component 4,(methylnitrosamino),1,(3,pyrodyl),1,butanone (NNK). Methods HPV,18 immortalized human bronchial epithelial cells (BEP2D) were used as the target cells. Cigarette smoke condense (CSC) was collected from a famous brand cigarette (mixed cigarette, the amount of tar were marked by 8 mg per cigarette), and the content of NNK in the CSC was detected by GC,TEA. A rapid and sensitive multinuclear cell assay was used to detect the frequency of the HPRT gene mutation in BEP2D cells induced by NNK and CSC. Results Content of CSC in the cigarettes was (7.93±0.89) mg per cigarette, content of NNK in the CSC was (0.046±0.008)μg per cigarette. The frequency of HPRT gene mutation in BEP2D cells (y1 and y2) presented dependent on the concentrations of NNK and CSC(x1 and x2), and the dose,effect relationship were: y1=0.016x1+1.012 and y2=0.0061x2+1.042 respectively. However, result computed base on above formula indicated that, in a cigarette, the fraction of NNK,induced HPRT gene mutation was only 0.002% less than that of CSC induced. Conclusion Both CSC and NNK could induce HPRT gene locus mutation but the fraction of NNK in a cigarette was minor.