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目的探讨XPD基因在氢醌(HQ)致U937细胞DNA损伤修复中的作用。方法选择5、10、20、40、80μmol/L HQ分别对U937细胞染毒12、24、48 h,根据各浓度下U937细胞活性,选择10、20、40μmol/L HQ染毒24、48 h进行后续实验。取10、20、40μmol/L HQ染毒24、48 h U937细胞,分别设为HQ低、中、高浓度组,另取未染毒U937细胞作为空白对照组,采用单细胞凝胶电泳检测各组细胞尾矩(TM)、Oliv尾矩(OTM),采用Western blotting法检测各组XPD蛋白相对表达量。结果与空白对照组比较,HQ低、中、高浓度组染毒24、48 h TM和OTM均明显增大(P均<0.05);与同组染毒24 h比较,HQ低、中、高浓度组染毒48 h TM和OTM均明显增大(P均<0.05),染毒48 h HQ低、中、高浓度组TM和OTM依次升高(P均<0.05)。与空白对照组比较,HQ低、中、高浓度组染毒24 h XPD蛋白相对表达量均明显升高(P均<0.05),且HQ高浓度组明显高于HQ低浓度组(P均<0.05);染毒48 h,HQ高浓度组XPD蛋白相对表达量明显高于空白对照组及HQ低、中浓度组(P均<0.05),但空白对照组及HQ低、中浓度组两两比较差异均无统计学意义(P均>0.05);HQ各浓度组染毒48 h XPD蛋白相对表达量与染毒24 h比较差异均无统计学意义(P均>0.05)。结论 XPD基因表达上调可促进HQ致U937细胞DNA损伤修复。
Objective To investigate the role of XPD gene in the DNA damage repair of U937 cells induced by hydroquinone (HQ). Methods U937 cells were treated with 5, 10, 20, 40, 80μmol / L HQ for 12, 24 and 48 hours, respectively. According to the activity of U937 cells at various concentrations, 10, 20 and 40μmol / Follow-up experiment. U937 cells were treated with 10, 20 and 40μmol / L HQ for 24 and 48 h, respectively. The U937 cells were divided into HQ low, medium and high concentration groups. U937 cells were used as blank control group and single cell gel electrophoresis Group tail moment (TM) and Oliv tail moment (OTM) were measured. The relative expression of XPD protein in each group was detected by Western blotting. Results Compared with the blank control group, the levels of HQ in low, middle and high concentrations of 24, 48 h TM and OTM were significantly increased (all P <0.05). Compared with the control group, HQ low, middle and high The concentrations of TM and OTM increased significantly at 48 h after exposure (all P <0.05). The levels of TM and OTM in HQ low, medium and high concentrations group increased significantly after 48 h exposure (all P <0.05). Compared with the blank control group, the expression of XPD protein in HQ low, middle and high concentration groups were significantly increased 24 h after exposure (all P <0.05), and HQ high concentration group was significantly higher than HQ low concentration group (P < 0.05). After exposure for 48 h, the relative expression of XPD protein in HQ high concentration group was significantly higher than that in blank control group and HQ low and middle concentration groups (P <0.05) (P> 0.05). There was no significant difference in the relative expression of XPD protein between the HQ groups at 48 h and 24 h after exposure (P> 0.05). Conclusion Upregulation of XPD gene may promote the DNA damage repair of U937 cells induced by HQ.