摘要
目的研究薯蓣皂苷元(Diosgenin,Dio)诱导人胃低分化粘液腺癌(MGC-803)细胞死亡的机理。方法四唑蓝(MTT)比色法检测Dio对肿瘤细胞生长抑制作用;流式细胞仪检测Dio对MGC-803细胞膜Annexin V表达的影响;APO-BRDU试剂盒检测Dio对MGC-803细胞DNA的影响;酶联免疫分析仪测定Dio对MGC-803细胞caspase-3活力和caspase-3抑制剂Ac-DEVD-CHO对Dio诱导细胞死亡的影响。结果一定浓度范围内Dio对MGC-803细胞增殖有剂量、时间依赖性抑制作用;经Dio处理后Annexin V-FITC+/PI-的凋亡细胞逐渐增多;随Dio剂量增大,DNA断裂碎片增多,凋亡细胞也逐渐增多。Dio作用细胞24 h,明显增强细胞caspase-3的活性。caspase-3抑制剂Ac-DEVD-CHO能部分抑制MGC-803细胞的凋亡。结论 Dio通过caspase-3途径诱导人胃低分化粘液腺癌细胞凋亡。
Objective AIM To study the antitumor mechanism of Dio for human gastric adenocarcinoma MGC-803 cells.Methods MGC-803 cells viability and the effect of caspase-3 inhibitor（DEVD-CHO） on Dio-induced apoptosis were measured by MTT assay.Annexin V expression in MGC-803 cell membrane was assessed by a flow cytometer.The effect of Dio on DNA in MGC-803 cells was measured using APO-BRDU Kit.Caspase-3 activity was detected by a enzyme link immune detector.Results Dio inhibited MGC-803 cells growth in dose-and time-dependent manners.MGC-803 cells treated with Dio showed typical characteristics of apoptosis：Annexin V-FITC＋/PI-and DNA fragmentation.Caspase-3 in MGC-803 cells was activated by Dio.Caspase-3 inhibitor,Ac-DEVD-CHO partially inhibited Dio-induced apoptosis.Conclusion Dio-induced apoptosis is partially dependent on caspase-3 pathway in MGC-803 cells.
出处
《中国实验诊断学》
北大核心
2011年第11期1813-1815,共3页
Chinese Journal of Laboratory Diagnosis
基金
国家自然科学基金项目(31100250)
吉林省人力资源与社会保障厅博士后基金课题
