摘要
目的探讨多聚聚合酶-1(PARP-1)在氢醌(hydroquinone,HQ)诱导的DNA损伤应答中的作用及其相关的调控机制。方法以磷酸盐缓冲液(PBS)溶解HQ,以10μmol/L HQ染毒TK6细胞为处理组,分别在染毒0.5、1、2、3、4、5和6 h时收集细胞,以0 h为对照组,或用HQ处理PARP-1沉默细胞,运用western bloting技术检测TK6细胞中磷酸化组蛋白H2AX(γ-H2AX)、聚腺苷二磷酸核糖(PAR)以及DNA损伤应答(DDR)相关蛋白PARP-1和抗凋亡转录因子(AATF)的表达情况。结果 HQ处理TK6细胞后,γ-H2AX的含量在3 h时达到高峰[(14.83±1.14)倍](P<0.05),此后逐渐下降;AATF的含量一直上升,6 h含量最高[(15.62±1.14)倍](P<0.05)。PARP-1的含量先下降后上升,3 h其含量最低[(0.66±0.04)倍](P<0.05)。PAR的含量在0.5 h达高峰[(1.78±0.13)倍](P<0.05),而后逐渐下降。PARP-1沉默细胞中,HQ处理使得PARP-1、PAR和AATF的含量分别降了77%(P<0.05)、64%(P<0.05)和68%(P<0.05),γ-H2AX的含量上升了1.65倍(P<0.05)。结论 PARP-1通过聚多聚腺苷二磷酸(ADP)核糖基化作用增强AATF稳定性参与由氢醌诱导的DNA损伤修复。
[Objective]To investigate the action and related regulatory mechanisms of poly ADP-ribose polymerase-1（PARP-1）on hydroquinone（HQ）-induced DNA damage response.[Methods]Hydroquinone was dissolved with PBS buffer,10 μmol/L HQ was given to the TK6 cells and incubated for 0.5,1,2,3,4,5 and 6 hours respectively,and the cells without treatment was the control group. In the other way,PARP-1silent cells were treated by HQ to establish the model. The expression of γ-H2 AX,PAR DDR-related protein PARP-1 and AATF were detected by western blotting. [Results]After TK6 cells were treated by HQ,the expression of γ-H2 AX reached the peak at 3 hours [（14.83 ±1.14） fold]（P 〈0.05）,while decreased gradually after that. The expression of AATF has been rising and reached the peak at 6 hours [（15.62±1.14） fold]（P〈0.05）. The expression of PARP-1decreased and then increased,and hit the bottom at 3 hours[（0.66 ±0.04）fold]（P〈0.05）. The quantity of PAR reached the peak at 0.5 hour [（1.78±0.13）fold]（P〈0.05）,while decreased gradually after that. In PARP-1silent TK6 cells with treatment of HQ,the expression of PARP-1,PAR and AATF respectively decreased by 77%（P〈0.05）,64%（P〈0.05） and 68%（P〈0.05）,while the expression of γ-H2 AX increased by 1.65 fold（P〈0.05）. [Conclusion]PARP-1 involves in HQ-induced DNA damage repair by via enhancing AATF stability through the action of poly（ADP-ribosyl）ation.
出处
《职业与健康》
CAS
2015年第14期1906-1909,共4页
Occupation and Health
基金
国家自然科学基金资助(项目编号:81202231)
广东省科技计划项目(项目编号:2013B021800066)
