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目的探讨核转录因子-κB(NF-κB)及其抑制因子(IκB)在肿瘤坏死因子-α(TNF-α)诱导人脑胶质瘤细胞株U 251细胞凋亡过程中生物活性的变化。方法应用流式细胞仪检测TNF-α对U-251细胞生长的抑制和诱导凋亡作用。用免疫组化方法检测肿瘤细胞p65的表达,用W estern b lot检测IκB蛋白的变化。结果TNF-α可诱导U 251细胞凋亡,激活细胞中p65并诱导IκB降解;抗氧化剂二硫碳吡咯烷醇(PDTC)可抑制TNF-α诱导的IκB的降解及NF-κB的激活,增强TNF-α诱导U 251细胞的凋亡。结论TNF-α在诱导U 251细胞凋亡的过程中可激活NF-κB。抑制NF-κB活性,可增强TNF-α诱导细胞凋亡的作用。
Objective To investigate the changes of biological activity of nuclear factor-κB (NF-κB) and its inhibitory factor (IκB) in the process of apoptosis induced by tumor necrosis factor-α (TNF-α) in U 251 human glioma cell line. Methods The inhibitory effect of TNF-α on the growth of U-251 cells and the induction of apoptosis were detected by flow cytometry. The expression of p65 in tumor cells was detected by immunohistochemistry and the changes of IκB protein were detected by Western blot. Results TNF-α induced the apoptosis of U 251 cells and activated the expression of IκB in cells. Antioxidant dithiopyrrolidine (PDTC) inhibited the degradation of IκB and the activation of NF-κB induced by TNF-α TNF-α induces apoptosis in U 251 cells. Conclusion TNF-α can activate NF-κB during U 251 cell apoptosis. Inhibition of NF-κB activity, can enhance TNF-α-induced apoptosis.