PARP抑制剂5-AIQ对小鼠结肠癌CT26细胞PARP/NF-κB复合物和NF-κB活性的影响

Effect of PARP inhibitor 5-AIQ on PARP/NF-κB complex and NF-κB activity in murine colon carcinoma CT26 cells

目的探讨PARP抑制剂5-AIQ对小鼠结肠癌CT26细胞核内PARP抑制后,对PARP/NF-κBP65复合物形成和NF-κB活性的影响。方法Western blot检测结肠癌CT26细胞核内PARP与NF-κBP65的表达,免疫共沉淀法分析CT26细胞核内PARP/NF-κBP65复合物形成,电泳迁移率改变分析法检测CT26细胞核转录因子NF-κB的结合活性。结果5-AIQ处理组与5-AIQ未处理组比较,前者小鼠结肠癌CT26细胞核内PARP与NF-κBP65的表达水平均明显减弱(P<0.05),PARP/NF-κBP65复合物形成明显减少(P<0.05),NF-κB的活性显著降低(P<0.05)。结论5-AIQ可通过抑制PARP的表达,减少PARP/NF-κBP65复合物的形成,进而使NF-κB活性降低。

Objective To investigate the effect of PARP inhibitor 5-AIQ on PARP/NF-κB P65 complex and the NF-κB activity in murine colon carcinoma CT26 cells. Methods The mufine colon carcinoma CT26 cells were treated with 5-AIQ in vitro. The expressions of PARP and NF-κB P65 were analyzed by Western Blot. The PARP/ NF-κB P65 complex was analyzed by co-immunoprecipitation. The NF-κB binding activity was detected by electro- phoretic mobility shift assay （EMSA）. Results Compared with 5-AIQ-untreated group, the expressions of PARP and NF-κB P65 were markedly decreased in 5-AIQ-treated colon carcinoma CT26 cell groups （P 〈0. 05）. The PARP/NF-κB P65 complex was decreased in 5-AIQ-treated colon carcinoma CT26 cell group （P 〈0.05）. The NF-κB DNA binding activity was also reduced in 5-AIQ-treated colon carcinoma CT26 cell groups （P 〈 0. 05 ）. Conclusion 5-AIQ can inhibit PARP expression in murine colon carcinoma CT26 cells, so as formation of PARP/ NF-κB P65 complex and NF-κB activity.