WRN基因在氢醌致U937细胞DNA损伤中的作用

The Effect of WRN gene on DNA damage induced by hydroquinone in U937 cells

目的 WRN基因在氢醌(HQ)致U 937细胞DNA损伤中的作用。方法常规培养白血病细胞U 937至生长对数期,低剂量HQ组、中剂量HQ组、高剂量HQ组分别以10、20、40μmol/L HQ染毒24h及48h,以等体积的完全培养基培养的细胞组为完全空白对照组。采用单细胞凝胶电泳(SCGE)检测细胞DNA损伤;采用免疫印迹法检测WRN蛋白的相对表达量。结果 (1)HQ可导致细胞DNA损伤,且损伤效用随染毒浓度增加而增大,48h比24h的DNA损伤程度增加,呈时间—剂量依赖性(P<0.05);(2)免疫印迹结果显示,HQ染毒24h,WRN蛋白相对表达量在各组差异无统计学意义(P>0.05);HQ染毒48h高剂量组分别与空白组、低剂量、中剂量中比较,WRN蛋白相对表达量呈降低的趋势(P<0.05)。结论 HQ诱导WRN蛋白表达下调影响U 937细胞DNA损伤修复。

Objective To study the effect of WRN gene on DNA damage induced by hydroquinone （HQ） in U937 cells.Methods U937 cells were treated with HQ at concentrations of 10,20 and 40μmol/L for 24 h and 48 h,respectively.The blank control group cells were cultured in the same volume of medium.The DNA damage was detected by single cell gel electrophoresis （SCGE）.The relativeexpression of WRN protein was detected by western blotting （WB）.Results HQ can cause DNA damage in cells,and the damage effect increased with the increase of the concentration of HQ,48 h than 24 h of DNA damage increased in a time-dose-dependent（P〈0.05）.WB analysis was showed that the expression of WRN protein in HQ group there were not statistical significantly differences （P〉0.05）.Compared with blank group,low dose and middle dose group,the relative expression of WRN protein in high dose group was decreased at 48 h and there were statistical significantly differences （P〈0.05）.Conclusion HQ-induced down regulation of WRN protein expression in DNA repair of U937 cells.