吴茱萸碱作用mTOR信号通路引起胃癌SGC-7901细胞凋亡的研究  

Effect of evodiamine in inducing apoptosis of gastric cancer SGC-7901 cells through mTOR signal pathway

作　　者：刘鑫[1] 杨丽[2] 毕瑩[2] 王良宏[2] 黄海[2] 

机构地区：[1]贵州省人民医院检验科,贵州贵阳550004 [2]贵州医科大学医学检验学院临床生化教研室,贵州贵阳550004

出　　处：《中国中药杂志》2015年第16期3262-3266,共5页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81260658;81460364)

摘　　要：吴茱萸碱(evodiamine)是中药吴茱萸最主要的具有抗肿瘤活性的生物碱之一,研究初步探讨吴茱萸碱作用哺乳动物雷帕霉素靶蛋白(m TOR)信号通路诱导胃癌SGC-7901细胞凋亡的作用机制,为其抗肿瘤机制的探索及胃癌临床治疗奠定基础。采用MTT法观察吴茱萸碱单独作用对SGC-7901细胞及对人外周血单个核细胞(PBMCs)的细胞毒效应。Real-time PCR,Western blot分别检验吴茱萸碱单独或联合胱天蛋白酶抑制剂(z-VAD-fmk)作用后,m TOR,p70S6K和4EBP1基因表达以及m TOR和p-m TOR蛋白表达。结果表明,吴茱萸碱抑制SGC-7901细胞增殖存在时间剂量依赖性,且对人PBMCs无细胞毒作用。吴茱萸碱及联合z-VAD-fmk分别作用后,细胞变圆,漂浮于培养基中;与对照组比较,2种处理方法均能抑制m TOR,p70S6K和4EBP1基因表达,存在显著性差异,且与吴茱萸碱单独作用比较,联合z-VAD-fmk作用对基因表达抑制率加强;Western blot结果显示吴茱萸碱可以抑制m TOR和p-m TOR蛋白的表达无论是否有z-VAD-fmk的参与,且z-VAD-fmk阻断胱天蛋白酶作用后,抑制率增强。综上说明吴茱萸碱可能通过m TOR信号通路促进胃癌SGC-7901细胞凋亡。Evodiamine is one of the most important antitumor alkaloid from evodiamine. This study focused on the mechanism of evodiamine in inducing apoptosis of gastric cancer SGC-7901 cells through mammalian target of rapamycin （mTOR） signal pathway, in order to explore its antitumor mechanism and lay a foundation for clinical treatment of gastric cancer. The sole cytotoxic effect of evodia- mine on SGC-7901 cells and human peripheral blood mononuclear cells （PBMCs） was observed by MTr assay. After the cells were re- spectively intervened with single evodiamine or evodiamine combined with z-VAD-fmk, the gene expressions of mTOR, p70S6K and 4EBP1 were analyzed by real-time PCR, and the protein expressions of mTOR and p-mTOR were detected by western blot. The result showed that evodiamine inhibited the apoptosis of SGC-7901cells in a time-dependent manner, with no cytotoxic effect on human PB- MCs. After the respective intervention with single evodiamine or evodiamine combined with z-VAD-fmk, the cells became round and floated in medium. Compared with the control group, both treatment methods can inhibit mTOR, 4E-BP1 and pTOS6K gene expres- sions, with significant differences. Compared with single evodiamine, evodiamine combined with z-VAD-fmk showed a higher inhibitory rate in gene expression. According to the Western Blot result, evodiamine can inhibit the protein expressions of mTOR and p-mTOR re- gardless of the combination with z-VAD-fmk, with a higher inhibitory rate after z-VAI）-fmk blocked caspase. In conclusion, evodiaminemay promote the apoptosis of SGC-7901cells through mTOR signal pathway.

关 键 词：吴茱萸碱 mTOR信号途径 胃癌SGC-7901细胞 细胞凋亡 

分 类 号：R285[医药卫生—中药学]