互隔交链孢酚对NIH/3T3细胞中DNA聚合酶β的致突变作用  

Mutation effect of alternariol on DNA polymerase beta in NIH/3T3 cell line

作　　者：朱涵[1] 樊红琨[1] 章茜[1] 

机构地区：[1]郑州大学医学院生理学教研室,河南省郑州市450052

出　　处：《中国组织工程研究》2012年第15期2831-2834,共4页Chinese Journal of Tissue Engineering Research

基　　金：教育部科学技术研究重点项目(204084);项目名称:DNA聚合酶β在互隔交链孢霉诱发癌变中的表达及修复作用的研究~~

摘　　要：背景:近年来研究发现食管癌细胞中存在DNA聚合酶β的突变。目的:观察互隔交链孢酚对NIH/3T3细胞中DNA聚合酶β基因序列的影响,研究互隔交链孢酚致细胞变异的机制。方法:用不同浓度(2,4,6,8μmol/L)的互隔交链孢酚作用于NIH/3T3细胞,并设置对照组。用RT-PCR法从细胞总RNA中扩增出DNA聚合酶β基因cDNA序列,应用T-A克隆技术,克隆至pGEM-T载体后进行测序,分析其序列变化。结果与结论:2μmol/L互隔交链孢酚处理后的细胞内DNA聚合酶β基因序列无任何变化。而4μmol/L组中DNA聚合酶β基因有1个位点发生突变,8μmol/L组和16μmol/L组中各有2个位点发生突变,此3组的突变存在相同的位点。结果证实,互隔交链孢酚可导致NIH/3T3细胞中的DNA聚合酶β发生点突变,互隔交链孢酚浓度越高,点突变的数量越多,且存在突变活跃位点。BACKGROUND：The mutation of DNA polymerase β（DNA pol β） has been found in esophageal carcinoma cells.OBJECTIVE：To analyze the effect of alternariol on the sequence of DNA pol β in NIH/3T3 cell line and to study the mutation mechanism of alternariol METHODS：The NIH/3T3 cell line was injected with different concentrations of alternariol（2,4,6,8 μmol/L） and the control group was set.DNA pol β gene was amplified from the total RNA of the cells using reverse transcription-PCR,then cloned with pGEM-T vector and sequenced with T-A cloning technique,and the sequence was analyzed.RESULTS AND CONCLUSION：The sequence of DNA pol β was not changed in 2 μmol/L alternariol group,and there was one mutation point in 4 μmol/L alternariol group and two mutation points in 8 and 16 μmol/L alternariol group.There was a same mutation point in these three groups.Alternariol can induce point mutation of DNA pol β gene in NIH/3T3 cell line.Higher concentration of alternariol,more points of DNA pol β gene mutation.The active mutation point exists.

关 键 词：互隔交链孢酚 DNA聚合酶Β NIH/3T3细胞 克隆 突变 组织工程 

分 类 号：R318[医药卫生—生物医学工程]