摘要
目的:研究大黄素(Emodin)对人胃癌细胞MKN45的作用及探讨其诱导MKN45凋亡的分子机制。方法:用MTT法观察大黄素对MKN45的生长抑制作用;荧光染色法及流式细胞仪分析诱导凋亡作用;蛋白质印迹法检测p53和p21蛋白水平的变化;细胞免疫化学法检测Fas的表达。结果:与对照组相比,大黄素能明显抑制MKN45细胞的生长且呈浓度、时间依赖性,其IC50(48h)为(47.5±0.3)μmol/L;大黄素处理胃癌细胞后,吖啶橙(AO)染色观察示,细胞核内可见浓染致密的颗粒荧光、新月型改变、核固缩或片段化及凋亡小体的形成。流式细胞仪分析显示,大黄素能浓度依赖性诱导MKN45细胞凋亡和G2/M期阻滞。蛋白质印迹法检测显示,随着药物浓度的不断增加,p53和p21WAF1蛋白水平表现出明显增强的趋势;细胞免疫化学方法显示,大黄素处理后的细胞Fas/APO-1表达较对照组明显增多。结论:大黄素对人胃癌细胞MKN45有明显的生长抑制作用,且该作用与p53依赖性诱导凋亡以及Fas表达水平的上调有关。
OBJECTIVE： To study the apoptosis induced by emodin in human gastric carcinoma cell line MKN45, and clarify its molecular mechanism. METHODS： The growth inbitory effect of emodin on human gastric carcinoma cell line MKN45 was observed by using MTT assay, and cell apoptosis was analyzed by fluorescent microscopy and flow cytometry. The protein expressions of p53 and p21 affected by emodin were determined by Western blot, and Fas by immunocytochemistry. RESULTS： Emodin significantly inhibited cell growth of human gastric carcinoma cell line MKN45 with IC50 value（48 h） of （47.5 ± 0. 3）μmol/L. After incubated with emodin, typical morphological changes could be observed by using fluorescent microscope, including nuclear chromatin condensation, fragmentation, cell shrinkage, and formation of apoptosis bodies; apoptosis which correlated with concentration of emodin and G2/M phase blocking could be detected by flow cytometry; the expression levels of p53, p21 and Fas had changed remarkably. CONCLUSION： The apoptosis in human gastric carcinoma cell line MKN45 could be induced by emodin in this study, the molecular mechanism may be p53 dependent, and Fas may also play important roles in the apoptosis process.
出处
《中华肿瘤防治杂志》
CAS
2011年第12期917-920,共4页
Chinese Journal of Cancer Prevention and Treatment
基金
甘肃省科技支撑计划项目(0804NKCA095)
甘肃省卫生行业科研计划管理项目(GWGL2010-9)
关键词
大黄素
胃肿瘤
凋亡
P53
emodin
stomach neoplasms
apoptosis
p53
