【摘要】 目的:建立稳定表达bcr/ablP210融合蛋白的小鼠转化细胞株BaF3-P210的DNA损伤模型,探讨bcr/abl融合蛋白对细胞DNA损伤修复功能的影响.方法:彗星实验检测不同浓度H2O2诱导的BaF3-P210细胞的DNA损伤建立DNA损伤模型,以空载体转染BaF3-MIGR1细胞为对照;用彗星实验对两组细胞在DNA损伤后的修复进行不同时间段的动态观测.结果:彗星实验确定了细胞DNA损伤模型的建立条件是:采用50μmol/L终浓度的H2O2对BaF3-MIGR1细胞和BaF3-P210细胞进行染毒,H2O2作用温度和时间均为4℃,10min.BaF3-P210细胞与BaF3-MIGR1细胞相比DNA损伤后修复时间缩短,在损伤后60min即可达到完全修复,而后者在损伤后120min才能完全修复(P<0.05).结论:建立了表达bcr/ablP210融合蛋白的小鼠转化细胞株BaF3-P210细胞的DNA损伤模型,并发现bcr/abl融合蛋白可增强BaF3-P210细胞的DNA损伤后修复能力.更多还原

【Abstract】 AIM:To establish the DNA damage model for the transformed mouse BaF3-P210 cell line stably expressing bcr/abl and to investigate the influence of bcr/abl fusion protein on the cell repair function after DNA damage.METHODS:The comet assay was employed to detect the DNA damage in BaF3-P210 cells induced by hydrogen peroxide at various concentrations to construct the DNA damage model.The BaF3-MIGR1 cell line transfected by the empty vector MIGR1 without the bcr/abl fusion gene was used as a control.A dynamic observation on the DNA repair after cell DNA damage was performed by the comet assay.RESULTS:The cell DNA damage models for both BaF3-P210 and BaF3-MIGR1 cells were established with hydrogen peroxide treatment at a final concentration of 50 μmol/L for 10 min at 4℃ as confirmed by the comet assay results.When compared with the control BaF3-MIGR1 cell group,the time duration required for the DNA repair of the BaF3-P210 cell group was significantly decreased,with 60 mins for BaF3-P210 cells and 120 min for BaF3-MIGR1 cells(P<0.05).CONCLUSION:The cell DNA damage model for the transformed mouse BaF3-P210 cell line stably expressing bcr/abl is successfully established and bcr/abl fusion protein strengthens the DNA repair capacity of the BaF3-P210 cell line after DNA damage.更多还原