彗星试验检测甲氨喋呤诱发人淋巴细胞DNA的损伤

目的用彗星试验评价人外周血淋巴细胞暴露于不同剂量甲氨喋呤（MTX）后的DNA损伤。方法外周血来自男女两名助血员,淋巴细胞分离后用终浓度为0（溶剂对照）、0、5、10、25、50、100ug ml^-1的MTX分别染毒6h和24h,而后用彗星试验检测各组淋巴细胞的DNA损伤,以尾长（TL）和尾相（TM）作为DNA损伤的指标,并分析剂量-效应关系。结果染毒组DNA损伤明显高于对照组,并有剂量-效应关系。结论彗星试验是一种快速、简便测定DNA损伤的方法。MTX对体外培养的人淋巴细胞具有DNA损伤效应。

Detecting DNA Damage of Human Lymphocytes induced by Methotrexate with Comet Assay

Objective To assess the DNA damage of human lymphocytes exposed to me thotrexate （MTX） at different doses. Methods Lymphocytes were separated from peripheral blood taken from two healthy donors （one male and another female）. Then the lymphocytes were incubated for 6 and 24 hours with 0 （solvent control group）, 5, 10, 25, 50, 100μg/ml MTX respectively. The Comet assay was used to detect the tail length （TL） which was served as a indicator of DNA damage as well as tail moment （TM）. Meanwhile the dose-effects relationship was analyzed. Results The rate of DNA damage of MTX- exposed group was significantly higher than that of control group. In addition, a dose-effects relationship was found. Conclusion Comet assay is a rapid and simple test to assess DNA damage. The result of this study indicates that MTX can induce DNA damage of human lymphocytes in vitro.