单细胞凝胶电泳法分析指标及检测DNA交联的标准断裂剂的研究  

Study on the parameters in single cell gel electrophoresis and hydrogen peroxide as a standard DNA breakage agent

作　　者：任泽舫[1] 庄志雄[2] 张锦周[2] 黄海雄[2] 张丙尧[2] 柳青[1] 

机构地区：[1]中山医科大学公共卫生学院流行病学教研室,广州510080 [2]中国预防医学科学院深圳研究中心毒理科

出　　处：《中华劳动卫生职业病杂志》2000年第4期210-212,共3页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：国家自然科学基金!(399706 36);深圳市科学技术计划!(199805033)资助项目

摘　　要：目的　寻求单细胞凝胶电泳计算机成像与分析系统中的有效分析指标 ,为H2 O2 在 1℃条件下作为检测DNA交联的标准DNA断裂剂提供依据。方法　常规培养Vero细胞 ,消化后悬浮在PBS中 ,H2 O2 染毒 ,分别在 37℃和 1℃条件下孵育 1h ,常规单细胞凝胶电泳 ,专用软件分析其图像。结果　H2 O2 的染毒浓度在 2 5 0 0 μmol/L以下时 ,尾长、尾面积、尾矩和尾积分等指标均有剂量 -效应关系 (T =37℃时 ,r分别为 0 .6 80 ,0 .72 5 ,0 .796 ,0 .798;T =1℃时 ,r分别为 0 .6 85 ,0 .717,0 .6 2 2 ,0 .714,P均 <0 .0 1)。在H2 O2 浓度较高的另一实验中 ,当H2 O2 为 2～ 7μmol/L时 ,仅尾矩和尾积分间存在这种关系 ;H2 O2 各剂量 (2 0、10 0、5 0 0、2 5 0 0 μmol/L)在 37℃时对DNA的损伤程度均明显高于1℃时。两种条件下上述 4个指标都存在剂量 -效应关系。结论　由该系统得到的尾矩和尾积分所反映的DNA损伤信息更准确、更全面、适用范围更广 ,但尾长可用作较初步的研究 ;H2 O2 在Objective To seek for the effective parameters in the imaging and analyzing system of single cell gel electrophoresis(SCGE),so as to provide basis regarding hydrogen peroxide at 1 ℃ as a standard DNA breakage agent.Methods Vero cells cultured by routine assay were digested and suspended in PBS,exposed to H 2O 2,incubated at 37 ℃ and 1 ℃ respectively for 1 hour.After SCGE,the images were analyzed with a special software for SCGE.The effect of H 2O 2 on DNA lesion at different temperatures and all the parameters were analyzed.Results Tail length,tail area,tail moment and tail integral could reflect DNA lesion extent as the concentration of H 2O 2 was below 2 500 μmol/L while the concentration was 2～7 μmol/L,just tail moment and tail integral could.The DNA lesion caused by H 2O 2 at 37 ℃ was significantly more severe than that at 1 ℃,but there were dose effect relationships at both temperatures. Conclusion Tail moment and tail integral were more accurate and suitable for extensive use whereas tail length could be used in rough analysis.H 2O 2 at 1 ℃ may be regarded as a standard DNA breakage agent to detect DNA crosslinks.

关 键 词：单细胞凝胶电泳法 分析指标 H2O2 DNA损伤 

分 类 号：R361[医药卫生—病理学]