摘要
AIM: To investigate the role of the mitochondrial pathway in JTE-522-induced apoptosis and to investigate the relationship between cytochrome C release, caspase activity and loss of mitochondrial membrane potential (Deltapsim). METHODS: Cell culture, cell counting, ELISA assay, TUNEL, flow cytometry, Western blot and fluorometric assay were employed to investigate the effect of JTE-522 on cell proliferation and apoptosis in AGS cells and related molecular mechanism. RESULTS: JTE-522 inhibited the growth of AGS cells and induced the apoptosis. Caspases 8 and 9 were activated during apoptosis as judged by the appearance of cleavage products from procaspase and the caspase activities to cleave specific fluorogenic substrates. To elucidate whether the activation of caspases 8 and 9 was required for the apoptosis induction, we examined the effect of caspase-specific inhibitors on apoptosis. The results showed that caspase inhibitors significantly inhibited the apoptosis induced by JTE-522. In addition, the membrane translocation of Bax and cytosolic release of cytochrome C accompanying with the decrease of the uptake of Rhodamin 123, were detected at an early stage of apoptosis. Furthermore, Bax translocation, cytochrome C release, and caspase 9 activation were blocked by Z-VAD.fmk and Z-IETD-CHO. CONCLUSION: The present data indicate a crucial association between activation of caspases 8, 9, cytochrome C release, membrane translocation of Bax, loss of Deltapsim and JTE-522-induced apoptosis in AGS cells.
瞄准：在 JTE-522-induced apoptosis 调查 mitochondrial 小径的角色并且调查在细胞色素 C 版本， caspase 活动和 mitochondrial 膜潜力(Deltapsim ) 的损失之间的关系。方法：房间文化，数的房间， ELISA 试金， TUNEL，流动 cytometry，西方的污点和 fluorometric 试金被采用在 AGS 房间和相关分子的机制在房间增长和 apoptosis 上调查 JTE-522 的效果。结果：JTE-522 禁止了 AGS 细胞的生长并且导致了 apoptosis。Caspases 8 并且 9 在从 procaspase 和 caspase 活动由劈开产品的外观判定了劈开特定的 fluorogenic 底层的 apoptosis 期间被激活。阐明是否 caspases 的激活 8 和 9 为 apoptosis 正式就职被要求，我们在 apoptosis 上检验了 caspase 特定的禁止者的效果。结果证明 caspase 禁止者显著地禁止了 JTE-522 导致的 apoptosis。另外，与 Rhodamin 123 的举起的减少伴随的细胞色素 C 的 Bax 和 cytosolic 版本的膜 translocation，在 apoptosis 的一个早阶段被检测。而且， Bax translocation，细胞色素 C 版本，和 caspase 9 激活被 Z-VAD.fmk 和 Z-IETD-CHO 堵住。结论：现在的数据显示在 caspases 的激活之间的一个关键协会 8， 9，细胞色素 C 版本， Bax 的膜 translocation， Deltapsim 的损失和在 AGS 房间的 JTE-522-induced apoptosis。
作者
Hong-Liang Li Xiao-Hong Li Jun-Hua Lü Xian-Da Ren,Department of Pharmacology,Jinan University Pharmacy College,Guangzhou 510632,Guangdong Province,China Dan-Dan Chen,Department of Cardiology,First Affiliated Hospital,Zhongshan University,Guangzhou 510089,Guangdong Province,China Hai-Wei Zhang,Department of Pathology,Jinan University Medical College,Guangzhou 510632,Guangdong Province,China Cun-Chuan Wang,Department of laparoscopic surgery,First Affiliated Hospital,Jinan University Medical College,Guangzhou 510632,Guangdong Province,China
基金
National Natural Science Foundation of China,No.39770300,30070873
the Overseas Chinese Affairs Office of the State Council Foundation,No.98-33
