彗星分析技术检测辐射和化学物质诱导的DNA损伤

MEASUREMENT OF DNA DAMAGES INDUCED BY RADIATION AND CHEMICALS USING COMET ASSAY

目的 :用彗星分析技术(cometassay)检测邻苯二胺(o_phenylenediamine,o_PDA)和 60Coγ_射线对CHL细胞DNA的损伤。方法 :CHL细胞经0、2、4、6、8μmol/L的邻苯二胺染毒和0、2、4、6、8、10、15Gy的 60Coγ_射线照射后 ,进行单细胞凝胶电泳 ,测定彗星尾长。采用Origin4.2软件建立两种诱变剂的剂量与CHL细胞彗星尾长的剂效关系。 结果 :DNA的损伤程度(以彗星尾长为指标)随 60Coγ_射线剂量的加大和o_PDA浓度的增加而增强 ,CHL细胞彗星尾长(TL)与 60Coγ_射线的剂效关系符合线性平方模型 :TL=20.41 +2.42D +0.38D2;与o_PDA浓度的剂效关系符合线性平方模型 :TL=1.90 +1.46C +0.52C2。 结论 :60Coγ_射线剂量和o_PDA的浓度与细胞DNA损伤程度具有剂效关系 ;

Purpose: To study DNA damages induced by 60 Co γ _r adiation and o_phenylenediamine(o_PDA) using comet assay. Methods: The DNA damages induced by 0, 2, 4, 6, 8, 10, 15 Gy 60 Co γ_rays and 0, 2, 4, 6, 8 μmol / L o_PDA were measured using comet assay. The dose_respose relationship of the CHL cell t ail length and the dose of the two type of mutagen were established with Origin 4 software. Results: The tail length o f comet, index of DNA damage in this experiment, increased with the dose of 60 Co γ_ray and concentration of o_PDA, resp ectively. The dose_respose relationship of the CHL cell tail l ength(TL)and the dose of 60 Co γ_ray fitted for linear_quadratic model: TL=20.41+2.42D+0.38D 2; The dose_response relationships fo r DNA damage induced by o_PDA fitted for linear_quadratic mode l: TL=1.90+1.46C+0.52C 2. Conclusion: The comet assay is a use ful tool for determining the genotoxicity of environmental agent s.