摘要
目的:探讨三萜类中药成分熊果酸(UA)诱导乳腺癌细胞MCF-7的凋亡作用,并通过分析UA作用后MCF-7细胞的caspase-3和多聚ADP核糖多聚糖(PARP)蛋白表达的变化,探讨其诱导MCF-7细胞凋亡的分子机制。方法:采用细胞培养技术,用不同浓度的药物在一定的时间内处理细胞株,采用MTT法、活细胞原位光镜和荧光染色技术、流式细胞技术(FCM)、荧光免疫组织化学技术,图象分析技术,研究UA对caspase-3和PARP蛋白表达的影响,诱导细胞凋亡的作用。结果:UA剂量依赖性的抑制MCF-7细胞增殖,半数生长抑制剂量(IC50)为(22.6±3.0)μmol.L-1,诱导caspase-3和PARP表达增加,使细胞呈现典型的凋亡形态学特征,核质浓集,有凋亡小体。结论:UA诱导MCF-7细胞凋亡,其机制涉及到caspase-3和PARP依赖性凋亡调节信号通路。
Objective： To study the effect of ursolic acid （UA）, apentacyclic triterpene acid, on MCF-7 cell apoptosis, and probable mechanism involved by detecting the expressions of caspase-3 and poly ADP-ribose polymerase（PARP） at protein level. Method： MCF-7 cells were cultured with different concentrations of UA. Growth inhibition of UA on MCF-7 cells was evaluated by MTT assay. Cell cycle and sub-G1 peak were performed by FCM. Morphologic changes of UA-treated cells were observed by light microscope. Apoptotic cells with condensed or fragmented nuclei were visualized by Ho 33258 staining by a fluorescence microscope（EX： U.V. ）. The protein expression of caspase-3 and PARP was analyzed by immunofluorescence cell staining （SABC-Cy3）. Result： 24 hours after UA treatment, inhibition of MCF-7 cell growth was concentration-dependent. The IC50 value for UA was （22.6± 3.0） μmol·L^-1. Cell cycle anaysis by FCM showed that 50μmol·L^-1of UA arrested MCF-7 cell cycle at G0～ G1 phase. Morphological changes of MCF-7 Cells exhibited many of the hallmark features of apeptosis, including ehromatin clumps and aggregation and DNA fragmentation. UA increased caspase-3 protein expression. Conclusion： The results suggest that UA evokes MCF-7 cell apoptosis is correlation with the up-regulation of caspase-3. Our study indicated that UA might be a potential Chinese medical component for breast neoplasm.
出处
《中国中药杂志》
CAS
CSCD
北大核心
2006年第2期141-144,共4页
China Journal of Chinese Materia Medica
