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目的:研究肿瘤特异性凋亡基因(apoptingene)在诱导人淋巴瘤细胞U937凋亡时,cJun氨基末端激酶(JNK)信号通路的作用。方法:用含有apoptin基因的真核表达载体,瞬时转染体外培养的人淋巴瘤细胞U937。采用流式细胞仪、Westernblot、台盼蓝染色及RTPCR等研究其在不同时间条件下诱导U937细胞凋亡的作用。结果:Apopin基因瞬时转染U937细胞48h,可出现明显的凋亡;而且凋亡细胞的数量与apoptin基因的转染时间有关。诱导后24h,U937细胞中出现磷酸化的JNK蛋白的表达,诱导后48h达高峰;而非磷酸化的JNK蛋白表达无明显变化。结论:Apoptin基因具有诱导U937细胞凋亡的作用。JNK信号通路的激活,可能在apoptin基因诱导肿瘤细胞凋亡过程中发挥着重要作用。
Objective: To investigate the role of apoptingene in inducing apoptosis of human lymphoma cell line U937 by cJun amino terminal kinase (JNK) signaling pathway. Methods: The human lymphoma cells U937 cultured in vitro were transiently transfected with the eukaryotic expression vector containing apoptin gene. Flow cytometry, Western blot, trypan blue staining and RTPCR were used to study the effects of different concentrations of U937 on apoptosis of U937 cells. Results: U937 cells were transiently transfected with Apopin gene for 48h, and significant apoptosis occurred. The number of apoptotic cells was related to the transfection time of apoptin gene. The expression of phosphorylated JNK protein appeared in U937 cells 24 h after induction, and peaked at 48 h after induction. However, the expression of non-phosphorylated JNK protein did not change significantly. Conclusion: Apoptin gene can induce the apoptosis of U937 cells. Activation of JNK signaling pathway may play an important role in apoptin gene-induced tumor cell apoptosis.