镍冶炼烟尘致CHL细胞DNA损伤的彗星试验  

Application of Comet Assay to Nickel Refining Fumes-Induced DNA Damage

作　　者：周春凌[1] 吴永会[1] 焦广宇[1] 

机构地区：[1]哈尔滨医科大学公共卫生学院,黑龙江哈尔滨150001

出　　处：《工业卫生与职业病》2005年第6期376-379,共4页Industrial Health and Occupational Diseases

基　　金：国家自然科学基金资助项目(30371205)

摘　　要：目的利用单细胞凝胶电泳技术(彗星试验)观察不同染毒浓度镍冶炼烟尘对中国仓鼠肺成纤维(Chinese Hamster Lung Fibroblast,CHL)细胞DNA的损伤及其程度,以研究其致癌机制。方法采用镍冶炼烟尘对CHL细胞进行染毒,设阴性对照组和阳性对照组,采用单细胞凝胶电泳技术,镜下观察不同染毒时间和不同染毒浓度对DNA的作用,并分析相应指标。结果通过对受损细胞拖尾比率、延伸的尾动差、尾DNA百分含量及尾长4个指标反映细胞DNA受损程度和受损特征指标的分析显示,镍冶炼烟尘对CHL细胞可造成DNA的损伤,各浓度染毒组与阴性对照组比较,差异均有非常显著性(P<0.01)。高浓度染毒组的作用明显强于低浓度染毒组,高浓度染毒组4 h细胞拖尾率为70.6,拖尾细胞延伸的尾动差为4.18±1.92,与低浓度染毒组比较,差异有显著性(P<0.05)。各浓度染毒组细胞染毒4 h时损伤最严重。结论利用彗星试验可以检测出不同染毒浓度镍冶炼烟尘对CLH细胞DNA所造成的损伤,损伤有剂量-反应关系,损伤程度和特征随作用时间不同而变化。Objective To observe the DNA damage induced by different doses of nickel refining fumes on CHL cells in order to study the mechanism of carcinogenesis. Methods Drip different doses of nickel refining fumes into in vitro cultivated CHL cells. Establish a negative controlled group and a positive controlled group. Use the technique of single cell micro gel electrophoresis to break and electrophoresis the damaged DNA. Observe the comet images by fluorescence microscope and detect the relative markers in order to explore the effect that the nickel refining fumes will have on CHL cells through different affected time and dose. Results Analyze tailed cell%, extent TM, tail DNA% and tail length which can tell the degree and character of the damaged cell DNA. The results show that the damage induced by nickel refining fumes is more serious than the negative controlled group （P〈0.01）, and the effect of high-dose group is obviously powerful compared with low-dose group （P〈0.05）. In 4 h-high-dose group, the percentage of tailed cells is 70.6, the extent TM is 4. 18±1.92, and they are obviously different from the low-dose group. The damage after affecting 4 h is the most serious among three doses groups. It might have recovery effect when the cells are affected more than 4 h. Conclusions The technology of comet assay could detect the DNA damage of CHL cells induced by different doses of nickel refining fumes. The damage is related with the dosages. The degree and character would differ with the affected time.

关 键 词：彗星试验 镍冶炼烟尘 DNA损伤 

分 类 号：R114[医药卫生—卫生毒理学]