Objective To study the inhibition mechanism of elemene on proliferation of rat C6 glioblastoma cells in vitro. Methods Effect of elemene on the proliferation of C6 glioblastoma cells, the level of Hsp90 combined to Raf-1 and expression of Hsp90 ,Raf-1 and ERK protein were tested by cell-counting,co-immunoprecipitation assay and Western blotting analysis at different concentrations or time points,respectively. Results The proliferation of C6 glioblastorna cells was inhibited by elemene in a dose/time-dependent manner. The level of Hspg0 combined to Raf-1 and the expression of phosphorylated Raf-1 and ERK were down-regulated,but the level of total Hspg0 in C6 cells had no change. Conclusion Elemene can inhibit the proliferation of C6 glioblastoma cells in vitro, disrupt the molecular chaperone function of Hspg0,decrease the recruitment of HspgO in the Hsp90/Raf-1 complex,block the activation of Raf-1,lead to down-regulation of ERK pathway, and these may be the mechanism of anti-proliferation effect of elemene.%目的 探讨莪术提取物榄香烯抑制胶质瘤细胞体外增殖的机制.方法 采用细胞计数、免疫共沉淀、Western印迹等方法,分别检测不同浓度、不同作用时间榄香烯对C6胶质瘤细胞的增殖影响、对C6细胞中与Raf-1结合形成分子复合体的Hsp90水平的影响、对C6细胞Hsp90、Raf-1、ERK蛋白质表达的影响.结果 榄香烯对大鼠C6胶质瘤细胞具有明显的抑制增殖作用,该增殖抑制效应呈剂量、时间依赖性.榄香烯可明显下调C6细胞中与Raf-1结合的Hsp90水平及C6细胞的磷酸化Raf-1、ERK表达,但不影响总Hsp90的表达水平.结论 榄香烯能有效抑制胶质瘤细胞的体外增殖,破坏Hsp90的分子伴侣功能,阻碍Hsp90/Raf-1复合体的形成,使Raf-1不能被活化,最终下调胶质瘤细胞赖以生存的Raf/MEK/ERK通路,这可能是榄香烯抑制C6细胞增殖的机制.
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