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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.
OBJECTIVE: To establish a DNA damage model of mouse transformed cell line BaF3-P210 stably expressing bcr / ablP210 fusion protein and investigate the effect of bcr / abl fusion protein on DNA damage repair function.Methods: The expression of BaF3 induced by different concentrations of H2O2 -P210 cell DNA damage model was established, empty vector-transfected BaF3-MIGR1 cells as a control; comet assay of the two groups of cells after DNA damage repair at different time periods of dynamic observation.Results: comet assay to determine The cell DNA damage model was established under the following conditions: BaF3-MIGR1 cells and BaF3-P210 cells were exposed to 50μmol / L final concentration of H2O2, the temperature and time of H2O2 exposure were 4 ℃ and 10min, The repair time of MIGR1 cells was shorter than that of MIGR1 cells and completely recovered 60 minutes after injury, while the latter could be completely repaired 120 minutes after injury (P <0.05) .Conclusion: The mouse transformed with bcr / ablP210 fusion protein was established Cell strain BaF3-P210 cells and found that the bcr / abl fusion protein can enhance the DNA repair capacity of BaF3-P210 cells.