摘要
目的探讨β-榄香烯通过Fas/Fas L信号通路对胶质瘤细胞增殖和凋亡的影响。方法以人胶质母细胞瘤U87细胞系为研究对象,分为空白对照组和β-榄香烯实验组。采用细胞毒性试验分析β-榄香烯对U87胶质瘤细胞的细胞毒性作用;四甲基偶氮唑蓝(MTT)比色法检测β-榄香烯对胶质瘤的增殖能力的影响;流式细胞仪(FCM)及Caspase活性试剂盒检测细胞凋亡;流式细胞术检测胶质瘤细胞周期;实时定量PCR、Western blot法检测细胞内Fas、Fas L的表达水平。结果胶质瘤细胞系的细胞毒性作用随着β-榄香烯浓度增加而增强,IC50为45.73μg/m L;流式细胞仪分析β-榄香烯作用于U87细胞,阻滞周期于G1/S期;β-榄香烯作用于U87胶质瘤后Fas和Fas L的mRNA和蛋白表达水平上升;β-榄香烯作用于胶质瘤细胞有抑制增殖作用,并呈浓度和时间依赖性。结论β-榄香烯可能通过上调Fas/Fas L通路的表达水平,抑制胶质瘤细胞增殖并诱导凋亡。
Objective To explore the effect of β-elemene on glioma cells proliferation and apoptosis by the Fas/FasL signaling pathway. Methods U87 cells was divided into two groups: the blank control group and β-elemne treatment group. Cytotoxicity assay was used to measure the cytotoxicity of β-elemene on the glioma U87 cells. The effect of β-elemene on cell prolifer- ation was detected by MTT assay. Cell apoptosis was detected by Caspase activity assay, and the change of glioma apoptosis and cell cycle were detected by flow cytometry and fluorescence microscope. The influence of β-elemene on the expression level of Fas and FasL were measured by Real-time quantitative PCR and Western blot. Results With the concentration of β-el- emene increasing, the cytotoxic effect of β-elemene was elevated in glioma cell line and the concertration of inhibited cell viability (IC50) was 45.73 μg/mL. β-elemene induced the cell cycle arrest of the U87 cells in the G1/S phase. The expression of Fas and FasL were increased by the treatment of 13-elemene in both mRNA and protein level. Furthermore, proliferation of U87 cells was impeded by 13-elemene in a time- and does-dependent manner. Conclusion β- elemene can effectively inhibit the proliferation of glioma cells, and the apoptosis induction is related with the upregulation of Fas/FasL expression.
出处
《哈尔滨医科大学学报》
CAS
2015年第4期300-304,共5页
Journal of Harbin Medical University
基金
国家自然科学基金资助项目(81372700)