Department of Pathology, Chongqing Medical University, Chongqing 400016, People's Republic of China.
Pathol Oncol Res. 2009 Sep;15(3):323-8. doi: 10.1007/s12253-008-9116-y.
To investigate effect of poly (ADP-ribose) polymerase inhibition on the proliferation of CT26 cells in vitro and the mechanism of this effect. CT26 cells were treated with a range of concentrations of 5-Aminoisoquinolin-1-one (PARP inhibitor) in vitro. MTT assays and flow cytometry were used to determine the proliferation and cell cycle distribution, respectively, of the cells. The expression of PARP-1 was investigated by Western blot. The binding of Nuclear Factor-kappaB to DNA was detected by electrophoretic mobility shift assay. The proliferation of CT26 cells was significantly inhibited by 5-AIQ induced PARP inhibition in a dose-dependent manner. Proliferation was inhibited by 17.5% at 100 microM 5-AIQ, by 27.6% at 500 microM 5-AIQ and by 39.9% at 1000 microM (P < 0.05). After treatment with 5-AIQ, the proportion of cells in G(0)/G(l) phases increased and the proportion of cells in S phase decreased. The expression of PARP-1 was attenuated in 5-AIQ-treated CT26 cells (P < 0.05) and the binding of NF-kappaB to DNA binding was similarly diminished (P < 0.05). These results suggest that PARP inhibition reduced the proliferation of CT26 cells in vitro and influences the cell cycle. This inhibition is mediated by inhibiting PARP-1, which then diminishes the activity of NF-kappaB.
为了研究聚(ADP-核糖)聚合酶抑制对 CT26 细胞体外增殖的影响及其作用机制。采用不同浓度的 5-氨基异喹啉-1-酮(PARP 抑制剂)处理 CT26 细胞。MTT 法和流式细胞术分别用于测定细胞的增殖和细胞周期分布。采用 Western blot 法检测 PARP-1 的表达。采用电泳迁移率变动分析检测核因子-κB 与 DNA 的结合。5-AIQ 诱导的 PARP 抑制可显著抑制 CT26 细胞的增殖,呈剂量依赖性。100 μM 5-AIQ 抑制增殖 17.5%,500 μM 5-AIQ 抑制增殖 27.6%,1000 μM 5-AIQ 抑制增殖 39.9%(P < 0.05)。用 5-AIQ 处理后,G0/G1 期细胞比例增加,S 期细胞比例减少。5-AIQ 处理的 CT26 细胞中 PARP-1 的表达减弱(P < 0.05),NF-κB 与 DNA 的结合也明显减弱(P < 0.05)。这些结果表明,PARP 抑制可减少 CT26 细胞的体外增殖,并影响细胞周期。这种抑制是通过抑制 PARP-1 介导的,从而降低 NF-κB 的活性。