去甲斑蝥素对人乳腺癌细胞系的凋亡诱导作用及bcl-2基因的表达(英文)

Norcantharidin induces apoptosis in human breast cancer cell line

目的：探讨去甲斑蝥素抗肿瘤作用的分子机制。方法：用 10μ g/ml去甲斑蝥素处理体外培养的人乳腺癌细胞系 MCF-7。处理后，在不同时间点，采用普通光镜、电子显微镜观察去甲斑蝥素对乳腺癌细胞的诱导凋亡现象。利用流式细胞仪分析凋亡细胞百分比，用蛋白印迹杂交方法对凋亡抑制基因 bcl-2的表达情况进行检测。结果：经 10μ g/ml去甲斑蝥素处理 12 h后，可观察到 MCF-7细胞变形、出泡，从培养瓶底脱离。细胞染色和电子显微镜可观察到染色质浓聚、边集，且随着药物作用时间的延长，凋亡细胞百分比逐渐增加。与对照组相比，凋亡抑制基因 bcl-2的表达降低。结论 :诱导肿瘤细胞凋亡可能是去甲斑蝥素抗肿瘤作用的分子机制之一。

Objective: The current study was designed to identify molecular mechanism of antitumor effect produced by NCTD(norcantharidin) in human breast cancer cell line. Methods: MCF-7 cells were grown in standard conditions. At the indicated time points after NCTD treatment, evaluation of apoptotic morphology by light microscopy, analysis of chromatin condensation by electron microscopy, determination of the percentage of apoptotic cells by flow cytometry and Western blot analysis for detecting of bcl-2 gene expression were conducted. Results: Differences in cellular morphology were observed between NCTD-treated and control MCF-7 cells. Cell shrinkage, cell budding and chromatin condensation were observed in the cells treated with 10μ g/ml NCTD for 12 hours. Percentage of apoptotical cells was increased while the expression of bcl-2 products was decreased with the increasing of incubation time. Conclusion: The results indicate that one of the major antitumor mechanisms of NCTD may be through inducing cell apoptosis which related to the regulation of bcl-2 gene expression.