交链孢酚对成纤维细胞中DNA聚合酶β的影响  

Effect of alternariol on DNA polymerase β in NIH/3T3 cell Line

作　　者：朱涵[1] 杨春[1] 赵伟达[2] 王书春[1] 章茜[1] 

机构地区：[1]郑州大学医学院生理学教研室,河南郑州450052 [2]河南大学生命科学院生物工程系09级

出　　处：《现代预防医学》2012年第19期5071-5073,共3页Modern Preventive Medicine

基　　金：教育部科学技术研究重点项目 (204084)

摘　　要：目的观察霉菌毒素的致癌机制是否与其影响了细胞内的基因修复系统相关。方法体外培养成纤维细胞NIH/3T3细胞系,在培养液中加入16μM的互隔交链孢酚作用后,提取细胞全RNA,进行半定量分析了解其表达量的变化,RT-PCR扩增DNA聚合酶β基因cDNA序列,克隆至T载体后测序,分析其序列变化。结果克隆成功并测序后发现有DNA聚合酶β基因序列的两个位点发生突变。同时,通过半定量分析发现DNA聚合酶β表达量逐渐上调。结论 AOH可导致NIH/3T3细胞中的DNA聚合酶β发生点突变﹑表达量上调。OBJECTIVE To analyze the mutation and expression of DNA polymeraseβ（DNApolβ） in NIH/3T3 cell line affected by alternariol（AOH）,and study on the mechanism of alternariol inducing cancer.METHODS Affected the cells by AOH,DNApolβ gene was amplified through RT-PCR from the cells,then cloned with pGEM-T vector and sequenced,analyzed the sequence to detect the mutation of DNApolβ gene,and amplified β-actin at same time,then detected expression account of DNApolβ gene.RESULTS There were 2 points mutations of DNApolβ gene in cell.The expression was increased affecting by AOH.CONCLUSION AOH can induce point mutation of DNApolβ gene,also expression of DNApolβ is increased according to AOH.

关 键 词：互隔交链孢酚 DNA聚合酶Β 测序 半定量PCR 

分 类 号：R34[医药卫生—基础医学]