Apoptotic effects of ginsenoside Rh₂ on human malignant melanoma A375-S2 cells  

人参皂苷Rh2对人黑色素瘤A375-S2细胞的促凋亡作用（英文）

作　　者：费晓方[1,2] 王本祥 田代真一[4] 李铁津 马吉胜 池岛乔[1,5] 

机构地区：[1]长春中医学院附属医院中日医学研究所 [2]吉林大学生命科学学院 [3]吉林天然药物研究所 [4]昭和药科大学病态科学教研室 [5]吉林大学化学系

出　　处：《Acta Pharmacologica Sinica》2002年第4期29-36,共页中国药理学报（英文版）

基　　金：Project supported by the National Natural Science Foundation of China, № 39770847; Japan China Medical Association (1998) and Human Science Foundation (1999).

摘　　要：AIM: To study the mechanism of ginsenoside-Rh2 (G-Rh2)-induced growth inhibition of A375-S2 cells. METHODS: A375-S2 cell viability and the effect of caspase inhibitors on G-Rh2-induced apoptosis were measured by crystal violet assay. Changes in cellular morphology were observed by phase-contrast microscopy. Apoptosis-specific nucleosomal DNA fragmentation was assayed by agarose gel electrophoresis. Cell cycle distribution was measured by flow cytometry. RESULTS: G-Rh2 inhibited the A375-S2 cell growth in concentration-and time-dependent manners. Caspase family inhibitor, z-Val-Ala-Asp-fluoromethylketone ( z-VAD-fmk ), caspase-3 inhibitor, z-Asp-Glu-Val-Asp-fluoromethyl-ketone (z-DEVD-fmk), and caspase-8 inhibitor, z-De-Glu-Asp-fluoromethylketone ( z-IETD-fmk ), partially inhibited G-Rh2-induced apoptosis. But caspase-1 inhibitor, Ac-Tyr-Val-Ala-Asp-chloromethyl-ketone (Ac-YVAD-cmk), did not antagonize G-Rh2 induced-cell death. CONCLUSION: G-Rh2 suppresses the growth of A375-S2 cells in vitro by inducing apoptosis. G-Rh2 induced apoptosis is partially dependent on caspase-8 and caspase-3 pathway in A375-S2 cells. Other apoptotic pathways might be also related to the induction ofapoptosis by G-Rh2.目的:研究人参皂苷G-Rh₂诱导人黑色素肿瘤细胞A375-S2凋亡的分子生物学机制.方法:用结晶紫染色的方法测定细胞的死亡率.用倒置显微镜观察细胞形态学的变化.用琼脂糖凝胶电泳检测核酸片断.用流式细胞仪检测细胞凋亡和细胞周期.结果:G-Rh₂抑制A375-S2细胞增殖并在20μmol/L可以诱导A375-S2细胞产生凋亡.Caspase抑制剂z-VAD-frnk（caspase家族）,z-DEVD-fmk（caspase-3）或z-IETD-fmk（caspase-8）能部分抑制细胞凋亡.但是Ac-YVAD-cmk（caspase-1）不能抑制A375-S2细胞凋亡.结论:G-Rh₂在体外抑制A375-S2细胞的增殖,通过细胞形态学和核酸片断分析,G-Rh₂能够诱导A375-S2细胞产生凋亡.这种作用是通过细胞内caspase一类半胱氨酸蛋白酶进行信号传导的.G-Rh₂对A375-S2细胞的周期没有影响.

关 键 词：GINSENG SAPONINS apoptosis CASPASES melanoma 

分 类 号：R73-36[医药卫生—肿瘤]