中文摘要：

NF—κB已被证明在肿瘤和炎症过程中起到至关重要的作用。因此，建立抑制NF-κB信号通路的药物筛选模型至关重要。利用荧光素酶报告基因技术与蛋白印迹技术分别探索TNFα处理浓度及时间对NF-κB报告基因表达和NF—κB抑制亚单位It〈Bα降解的影响，进而构建NF—κB信号通路抑制剂药物筛选模型。实验结果表明，0．01nmol／LTNFα作用24h即能刺激HEK293T细胞中NF—κB报告基因较高水平的表达，且其表达量与TNFα的剂量和处理时间呈正相关性；0．01nmol／LTNFα作用5min即能使Panc-28细胞中IκBα明显降解，20min～30min几乎降解完全，之后IκBα含量又开始增加。NF-κB阳性抑制剂藤黄酸验证表明NF-κB萤光素酶报告基因检测筛选体系和NF—κB抑制亚单位降解筛选体系两种体系稳定可行。结果证明，两种模型可以用于NF—κB信号通路抑制剂药物的筛选。

英文摘要：

NF-κB is known as a major transcription factor that plays an essential role in the development of inflammation and cancer. Therefore, it＇ s a promising strategy to set up a drug screening model by inhibiting NF-κB signaling pathway. The effects of the concentrations and processing time of TNFα were explored on the NF-κB-luc reporter gene expression and the degradation of IκBα via the luciferase assay and western blot assay. The results showed, the expression level of NF-κB-luc reporter gene, which directly related to the concentrations and processing time of TNFα, was highly enough by the stimulation of 0.01 nmol/L TNFα for 24 h in HEK293T cells. IκBα degraded notably at the density of 0.01 nmol/L after 5 min treatment with TNFα on the Panc-28 cancer cells, and almost completely degraded after 20 min - 30min. Both the NF-κB luciferase reporter gene screening model and NF-κB inhibitory subunit screening model are confirmed by the known NF-κB positive inhibitor gambogic acid. Taken together, these two models can be used to select related potential drugs which inhibit NF-κB signaling pathway.