摘要
目的：明确Bax-Bcl-2异源二聚体与NSAIDs诱导胃癌细胞凋亡的关系。方法：以NSAIDs诱导胃癌细胞凋亡,并通过丫啶橙(AO)染色、共聚焦显微镜、流式细胞术、TUNEL法加以证实。应用Western blot方法检测Bax、Bcl-2蛋白的表达,应用免疫沉淀-蛋白印迹法检测Bax-Bcl-2异源二聚体水平的改变。结果：NSAIDs药物吲哚美辛(indomethacin,indo)800 mmol/L和阿司匹林(aspirin,Asp)8 mmol/L作用24 h后,AGS细胞发生显著的凋亡(Indo 800 mmol/L作用24 h凋亡率(9．34±1．99)%,48 h(38．97±3．36)%,Asp 8 mmol/L 48 h凋亡率(17．60±3．30)%。随着药物作用时间的延长,Bax-Bcl-2异源二聚体水平逐渐增高,在6～48 h内均呈现增强趋势,Bax蛋白表达的增强在6～24 h最为明显,Bcl-2蛋白未检测到。结论：NSAIDs可诱导胃癌细胞AGS凋亡；Bax-Bcl-2异源二聚体可能具有促进细胞凋亡的作用,也可能是NSAIDs调控肿瘤细胞凋亡的一个重要作用点。
Objective：To elucidate the relationship between Bax/Bcl-2 heterodimer and apoptosis of gastric cancer cells induced by non-steroidal anti-inflammatory drugs （NSAIDs）. Methods： Apoptosis was induced by NSAIDs and determined by acri dine orange （AO） staining, laser confocal laser scanning microscopy, flow cytometry, and TdT-mediated dUTP nick end labeling assay. The expression of Bax and Bcl-2 protein was detected by Western blot. The level of Bax-Bcl-2 heterodimer was determined by immuno-precipitation/Western blot （Ip-Wb） analysis. Results： Apoptosis of AGS cells were induced after 24-h treatment with indomethacin 800 mmol/L or aspirin 8 mmol/L. The apoptotic rate induced by indomethacin was （9.34±1.99）% at 24 h and （38. 97±3.36）% at 48 h. The apoptotic rate induced by aspirin was （17. 60±3.30）% at 48 h. The level of Bax-Bcl- 2 heterodimer gradually increased from 6 h to 48 h in a time-dependent manner. The increase in Bax protein was significant during 6-24 h. But the expression of Bcl-2 protein was undetected. Conclusion： NSAIDs could induce apoptosis of AGS cells. Bax/ Bcl-2 heterodimer might promote the apoptosis and serve as an important target for NSAIDs to regulate apoptosis.
出处
《肿瘤》
CAS
CSCD
北大核心
2006年第11期1002-1006,共5页
Tumor
基金
上海市科委自然科学基金(编号:04ZR14065)
