榄香烯对大鼠胶质瘤C6细胞Bcl-2家族基因及蛋白表达的影响  

Influence of elemene on the expression of Bcl-2 family genes in rat C6 glioma cells

作　　者：徐英辉[1] 董斌[1] 罗其中[2] 周洪语[2] 贾宜昌[3] 杨友峰[3] 王以政[3] 

机构地区：[1]大连医科大学附属第一医院神经外科,116011 [2]上海第二医科大学附属仁济医院神经外科 [3]中国科学院上海神经科学研究所

出　　处：《中华医学杂志》2005年第24期1700-1703,共4页National Medical Journal of China

基　　金：国家自然科学基金资助项目(30070769)

摘　　要：目的探讨莪术提取物-榄香烯诱导大鼠胶质瘤C6细胞系凋亡机制及其对Bcl2家族基因及蛋白质表达的影响。方法应用RTPCR、Westernblot方法检测榄香烯在0、20、40、60、80μg/ml浓度及作用时间为12、24、36、48、72h时处理的大鼠胶质瘤C6细胞系,利用流式细胞仪观察细胞的凋亡、细胞增殖的变化和Bcl2家族基因及蛋白质表达的变化。结果经榄香烯20、40、60、80μg/ml处理大鼠胶质瘤C6细胞48h后,单视野细胞计数分别为(536±9)个、(375±10)个、(246±9)个、(112±10)个,与浓度为0μg/ml时单视野细胞计数(625±12)个相比体外增殖抑制效应比差异有统计学意义(F=1292.416,P<0.05)。经不同浓度榄香烯处理后C6细胞凋亡率分别增加到(27±2)%、(29±4)%、(32±3)%、(35±5)%,同时有亚二倍体凋亡峰,即Ap峰出现。榄香烯可明显下调Bcl2/Bclx/l基因及蛋白质表达,且该作用呈浓度、时间依赖性,而对Bax基因及蛋白质无明显影响。结论下调Bcl2/Bclx/l基因及蛋白质表达可能是榄香烯诱导凋亡、抑制C6细胞增殖的主要机制。Objective To observe the effects of elemene on the induction of apoptosis in rat C6 glioma cells and its influence on expression of Bcl-2 family genes. Methods Rat C6 glioma cells were cultured. Elemene of the concentrations of 0, 20, 40, 60, and 80μg/ml were added for 12, 24, 36, 48, and 72 hours respectively. RT-PCR was used to detect the mRNA expression of Bcl-2/Bcl-x/1 genes. Western blotting was used to detect the protein expression of Bcl-2/Bcl-x/1 genes. The apoptosis of the cells was examined by flow cytometry. Results The cell counts of the 20, 40, 60, and 80 μg/ml elemene groups were 536±9, 375±10, 246±9, and 112±10/visual field respectively, all significantly lower than that of the 0 μg/ml elemene group (all F=1292.416,P<0.05) and the apoptotic rates of the 20, 40, 60, and 80 μg/ml elemene groups were (27±2)%,( 29±4)%,( 32±3)%, and (35±5)% respectively with an Ap peak. The protein expression of Bcl-2/Bcl-x/l genes was decreased in the elemene groups dose and time-dependently. The expression of Bax protein was decreased in the elemene groups too, however, not dose and time-dependently. Conclusion Apoptosis caused by elemene may be associated with the down-regulation of Bcl-2/Bcl-x/l genes.

关 键 词：BCL-2家族 榄香烯 胶质瘤 蛋白表达 Bcl-x/l基因 蛋白质表达 Western C6细胞系 增殖抑制效应 细胞增殖 细胞计数 莪术提取物 流式细胞仪 细胞凋亡率 Bcl-2 时间依赖性 Bax基因 凋亡机制 诱导大鼠 blot 作用时间 不同浓度 

分 类 号：R73-36[医药卫生—肿瘤]