中文摘要：

目的：利用细胞核亚细胞器结构的一些特异性蛋白质SC35、核孔复合体（nuclearporecomplex，NPC）、RNA聚合酶Ⅱ（PolⅡ）及Y12等标记亚细胞器，探讨转录抑制剂5，6-dichloro-1-β-D．ribofuranosylbenzimidazole（DRB）与仅．Amanitin对人非小细胞肺癌A549细胞中Nr￡2的表达与定位的影响。方法：①50mg/LDRB和2．5mg／L α-Amanitin分别作用于A549细胞1h和6h后，Westernblotting检测Nrf2及其相关蛋白的表达变化。②50mg/L DRB和2．5mg／L α-Amanitin分别处理A549细胞1h，利用免疫荧光细胞技术检测Nrf2、SC35、NPC、RNA PolⅡ、Y12形态结构的变化，并研究Nrf2定位的变化。结果：①与对照组相比，DRB和α-Amanitin作用1h后，Nrf2及其调控的下游基因HO-1、AKR1C和NQ01的蛋白质表达变化不明显，但6h作用后，这些蛋白质的表达明显降低。②激光共聚焦显微镜观察显示，细胞经DRB和α-Amanitin处理1h后，SC35的荧光强度明显增强，Pol11的荧光强度减弱，而Nrf2的定位无明显改变；NPC、Y12的表达无明显变化，且与Nrt2无共定位现象。结论：DRB和α-Amanitin能抑制Nrt2及其HO-1、AKR1C和NQ01的表达，但是对Nrf2的定位没有显著影响。

英文摘要：

Objective： To investigate the effects of transcriptional inhibitors 5,6-dichloro-1-b-D- ribofuranosylbenzimidazole （DRB） and α-Amanitin on the localization of Nrf2 in the nucleus. Methods： A549 cells were treated with DRB （50 mg/L） or α-Amanitin （2.5 mg/L） for 1 h and 6 h in serum-free medium,respectively. The expressions of Nrf2, HO-1, NQO1 and AKR1C were detected by Western blotting analysis. The localization of Nrf2 was determined by laser scanning confocal microscopy after cells were treated with either DRB or α-Amanitin for 1 h. Results： The expressions of Nrf2 and Nrf2-ARE gene batteries HO-1, AKR1C and NQO1 were decreased after 6 h treated with either DRB or α-Amanitin. The expression of SC35 was up-regulated but RNA Pol 11 was down-regulated; Y12 and NPC did not significantly change. The localization of Nrf2 in the cell nucleus did not change significantly. Conclusion： DRB and α-Amanitin can down-regulate the expression of Nrf2 and its targeting proteins HO-1 ,AKR1C and NQO1, but may have no effect on the localization of Nrf2.