论文部分内容阅读
目的 研究非离子型的diazeniumdiolate类一氧化氮供体引起肝癌细胞凋亡的分子机制。方法 利用免疫印迹、免疫沉淀、凝胶阻滞实验研究一氧化氮供体处理Hep3B肝癌细胞后,丝裂原激活的蛋白激酶、AP-1的激活以及和Hep3B肝癌细胞凋亡的关系。结果 一氧化氮可引起细胞外信号调节蛋白激酶、c-jun N末端激酶和p38激酶的激活,特别是细胞外信号调节的蛋白激酶的持续激活,其中细胞外信号调节的蛋白激酶和c-jun N末端激酶的特异的阻断剂U0126和JNK抑制剂Ⅱ可阻断AP-1的激活和Hep3B细胞的凋亡,而p38激酶的阻断剂SB203580不能阻断AP-1的激活和Hep3B肝癌细胞的凋亡。结论 一氧化氮通过激活细胞外信号调节蛋白激酶、c-jun N末端激酶,进而激活AP-1而引起Hep3B肝癌细胞的凋亡。
Objective To investigate the molecular mechanism of non-ionic nitric oxide (diazeniumdiolate) -induced hepatocarcinoma cell apoptosis. Methods Western blotting, immunoprecipitation and gel blocking assay were used to investigate the relationship between activation of mitogen-activated protein kinase (AP-1) and apoptosis of Hep3B hepatocarcinoma cells after treatment with nitric oxide donor. Results Nitric oxide induced the activation of extracellular signal-regulated protein kinase, c-jun N-terminal kinase and p38 kinase, especially the activation of extracellular signal-regulated protein kinase, including extracellular signal-regulated protein kinase and c-jun U0126, a specific inhibitor of N-terminal kinase, and JNK inhibitor II blocked the activation of AP-1 and the apoptosis of Hep3B cells. However, SB203580, a blocker of p38 kinase, failed to block AP-1 activation and Hep3B hepatoma cells Apoptosis. Conclusion Nitric oxide can induce apoptosis of Hep3B hepatoma cells by activating extracellular signal-regulated protein kinase, c-jun N-terminal kinase, and then activating AP-1.