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[目的]探讨苯并(a)芘[B(a)P]作用下人支气管上皮细胞16HBEDNA损伤与着色性干皮病A、C蛋白(XPA,XPC)表达的关系。[方法]用B(a)P(0、1、2、4、8、16、32、64μmol/L)浓度染毒细胞24h,用四甲基偶氮唑盐微量酶反应比色法(MTT)检测B(a)P对细胞活性的影响。用彗星试验检测细胞DNA损伤并以Olive尾矩值(OTM值)评价DNA损伤程度。并用Western-blot检测XPA和XPC蛋白的表达水平。[结果]B(a)P浓度>4μmol/L的各组细胞活性均显著降低(P<0.05)。各染毒组细胞DNA损伤程度则显著增高(P<0.05)。不同浓度B(a)P染毒细胞中XPA、XPC蛋白的表达量有差异。在1μmol/L B(a)P染毒组,可检出XPA蛋白表达水平增高,但随着染毒浓度增加其表达量逐渐降低(P<0.05)。与对照组相比各染毒组细胞XPC的表达均显著降低(P<0.05)。回归分析显示XPA与OTM值之间决定系数为0.572,XPC与OTM值之间为0.852。[结论]B(a)P引起的XPA和XPC蛋白表达水平的变化与DNA损伤之间存在高度相关。
[Objective] To investigate the relationship between the injury of human bronchial epithelial cells 16HBEDNA and the expression of protein A and protein C (XPA, XPC) in xeroderma pigmentosum by benzo [a] pyrene [B (a) P]. [Methods] The cells were exposed to B (a) P (0,1,2,4,8,16,32,64μmol / L) for 24 hours. MTT assay ) To examine the effect of B (a) P on cell viability. The DNA damage was detected by comet assay and DNA damage was evaluated by Olive tail moment (OTM). Western-blot was used to detect the expression of XPA and XPC protein. [Results] The cell viability of B (a) P group> 4μmol / L was significantly decreased (P <0.05). The DNA damage in all exposure groups was significantly increased (P <0.05). The expression of XPA and XPC in different concentration of B (a) P exposed cells were different. In the 1μmol / L B (a) P group, the expression level of XPA protein was increased, but the expression level of XPA protein decreased gradually with the increase of the concentration (P <0.05). Compared with the control group, the expression of XPC in all exposure groups was significantly decreased (P <0.05). Regression analysis showed that the coefficient of determination between XPA and OTM was 0.572 and that between XPC and OTM was 0.852. [Conclusion] The changes of XPA and XPC protein expression induced by B (a) P are highly correlated with DNA damage.