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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 investigate the role of WRN gene in DNA damage induced by hydroquinone (HQ) in U937 cells. Methods Normal leukemic cells U 937 were cultured in logarithmic phase, low dose HQ group, middle dose HQ group and high dose HQ group with 10, 20, 40μmol / L HQ for 24h and 48h, respectively. The cultured cell group was completely blank control group. The cell DNA damage was detected by single cell gel electrophoresis (SCGE). The relative expression of WRN protein was detected by Western blotting. Results (1) HQ could induce cell DNA damage, and the damage effect increased with the increase of the concentration of HQ. The DNA damage increased at a dose-dependent manner (P <0.05) at 48h compared with that at 24h. (2) The results showed that there was no significant difference in WRN protein expression between HQ groups (P> 0.05) and HN group (P> 0.05). Compared with the blank control group, low dose and middle dose groups, The amount showed a decreasing trend (P <0.05). Conclusion Down-regulation of WRN by HQ induces DNA damage repair in U937 cells.