真核细胞筛选系统快速检测环境水样的DNA损伤效应  

Screening DNA damage of organic extract in water sample of environment by eukaryotic cell screening system

作　　者：张荣[1] 杜海荣[2] 曹贤文[2] 周宜开[2] 

机构地区：[1]河北医科大学公共卫生学院劳动卫生与环境卫生学教研室,石家庄市050017 [2]华中科技大学同济医学院环境与健康教育部重点实验室

出　　处：《河北医药》2008年第5期581-583,共3页Hebei Medical Journal

基　　金：国家自然科学基金资助项目(编号:30471433);河北省科技厅科技支撑指导计划项目(编号:072761893)

摘　　要：目的利用含生长抑制和DNA损伤诱导基因153(gadd153)启动子和荧光素酶报道基因真核细胞株GADD153-Luc,建立快速检测环境水样中有机提取物的DNA损伤作用的新方法。方法RT-PCR方法检测内源性gadd153基因mRNA表达;生物发光和彗星试验分别检测环境水样对稳定转染细胞和HepG2细胞的DNA损伤效应。结果RT-PCR结果表明,在100ml/ml培养基剂量组,源水和氯化消毒饮用水中有机提取物可诱导gadd153基因mRNA表达显著增加(P<0.05),并与荧光素酶活性呈正相关;荧光素酶表达在源水和氯化消毒饮用水有机提取物各剂量组均显著高于对照组(P<0.01),并有良好的剂量反应关系;彗星试验显示,在10、100ml/ml培养基剂量组源水和氯化消毒饮用水有机提取物处理后,OTM(Olive尾距)显著高于对照组(P<0.01);相关分析表明,各剂量组诱导荧光素酶活性和DNA损伤程度(OTM)呈正相关(P<0.01)。结论真核细胞gadd153-Luc检测体系可用于快速检测环境水样的DNA损伤效应,并具有较高的灵敏性。Objective Constructing a hepatic cell line GADD153-Luc as eukaryotic cell screening system to rapidly detect the DNA damages of organic extract in water sample of environment. Methods Endogenic GADD153 mRNA level was detected by RT-PCR. DNA damage was detected by bioluminescence and comet assay. Results The high expression of gadd153 mRNA was induced by the organic extracts in water at dose of 100ml/ml medium （ P 〈 0.05）. There were positice correlation between the luciferase activity and the expression of gadd153 mRNA ; The luciferase activity was significantly increased in each treatment group at each dose（ P 〈 0.01 ）and in a dose-dependent manner; The Olive Moment （OTM） induced by raw water and chlorinated drinking water extracts was increased at dose of 10, 100ml/ml medium （ P 〈 0.01 ）, when compared with control group. There were positive correlations between the OTM and the luciferase activities. Conclusion Recombinant GADD153-Luc cell can rapidly detect DNA damage effect caused by water samples in environment and has a high sensitivity.

关 键 词：真核细胞筛选系统 源水和氯化消毒饮用水有机提取物 DNA-N 4h gadd153基因 荧光素酶报告基因 

分 类 号：R734.2[医药卫生—肿瘤] R123.1[医药卫生—临床医学]