苯并(a)芘致人支气管上皮细胞DNA损伤及与组蛋白H2A单泛素化水平的关系  

Correlation between DNA Damage and Expression of Mono-ubiquitinated Histone H2A in 16HBE Cells Exposed to Benzo[a]pyrene

作　　者：王志武[1] 贾利娜[1] 牛侨[1] 杨瑾[1] 

机构地区：[1]山西医科大学公共卫生学院劳动卫生学教研室,山西太原030001

出　　处：《环境与职业医学》2012年第3期136-138,143,共4页Journal of Environmental and Occupational Medicine

基　　金：国家自然科学基金项目(编号:30901180);山西省高等学校优秀青年学术带头人支持计划项目(编号:晋科教2010-4);太原市科技计划项目大学生创新创业专项(编号:201010-1390)

摘　　要：[目的]通过体外细胞培养途径,探讨苯并(a)芘(benzo[a]pyrene,B[a]P)作用下人支气管上皮细胞(16HBE)DNA损伤与组蛋白H2A单泛素化表达水平的关系。[方法]取对数生长期16HBE细胞,采用2μmol/L B[a]P染毒不同时间(0、1、2、4、8、12、24、48h)进行实验,用碱性单细胞凝胶电泳实验检测细胞DNA损伤水平,并以Olive尾矩值(olivetail moments,OTM)评价DNA损伤程度;用Western-blot法检测组蛋白H2A单泛素化水平。[结果]单细胞凝胶电泳结果显示:2μmol/L B[a]P染毒不同时间时,随着染毒时间的延长,各染毒组细胞DNA损伤程度均明显增高,与对照组相比,差异均有统计学意义(P<0.01);Western-blot结果显示:2μmol/L B[a]P染毒不同时间时,与对照组相比,组蛋白H2A的单泛素化水平增高,且在染毒2h后,差异有统计学意义(P<0.01);回归分析显示,单泛素化H2A的表达水平与DNA损伤程度存在高度相关,决定系数(R2)为0.910,P<0.01。[结论]B[a]P染毒致16HBE细胞DNA损伤与组蛋白H2A单泛素化水平之间存在密切关系。[Objective] To investigate the association between DNA damage and expression of mono-ubiquitinated histone H2A in human bronchial epithelial cells（16HBE） exposed to benzo[a]pyrene（B[a]P）.[Methods] The 16HBE cells collected at the logarithmic phase of growth were exposed to B[a]P at the concentration of 2 μmol/L for 0,1,2,4,8,12,24 and 48 h respectively.DNA damage was determined by comet assay and expressed as olive tail moments（OTM）.Protein expression was detected by Western-blot.[Results] The results of comet assay showed that the OTM values increased with exposure time and a significant increment was found as compared with the control group（P 0.01）.After exposed to 2 μmol/L B[a]P for 2 h,the mono-ubiquitination levels of histone H2A increased significantly when compared with the control group（P 0.01）.The regression analysis showed a significant correlation between OTM and the levels of mono-ubiquitinated histone H2A,and the determination coefficient was 0.910（P 0.01）.[Conclusion] The findings suggest a close correlation between DNA damage and the levels of mono-ubiquitinated histone H2A in 16HBE cells exposed to B[a]P.

关 键 词：苯并(A)芘 组蛋白H2A 人支气管上皮细胞株 DNA损伤 

分 类 号：R114[医药卫生—卫生毒理学]