人体颊黏膜细胞彗星实验方法学研究

Study on the Method of Human Buccal Cell Comet Assay

为探索一种简便、直接检测环境诱变物对人体靶细胞遗传毒性的方法,笔者以甲醛作为染毒剂,对人体口腔颊黏膜细胞彗星实验的染毒时间、染毒温度和2种用于检测交联作用的方案进行了研究。结果显示:经7 5μmol L甲醛37℃染毒30和60min后,DNA的断裂程度较染毒15min更大;而经该浓度甲醛在4,23和37℃下染毒30min后均可引起DNA的断裂,但37℃下染毒对DNA的断裂作用更大。在2种应用于检测交联作用的方案中,加大电泳条件的交联检测方案还可检测兼具交联和断裂效应的诱变物。

In order to establish a sensitive and rapid assay to test the genotoxicity of environmental genotoxin on human target cells,the authors carried out a study on the optimal duration and temperature of exposure treatment and two testing protocols for crosslinks.Results show that after exposed to 7.5 μmol/L of formaldehyde at 37 ℃ for 30 and 60 min,DNA was damaged more acutely than that for 15 min.After exposet to 7.5 μmol/L of formaldehyde at 4,23 and 37 ℃ for 30 min,DNA damage happened in all of the three groups,but at 37 ℃ DNA strands broke most.On the two testing protocols for crosslinks,increasing the duration of electrophoresis is more reasonable for the detecting of DNA breakage and crosslinks.