甘草查尔酮A对胶质瘤的增殖抑制及促凋亡作用机制研究

Licochalcone A inhibits proliferation and induces cell apoptosis in human glioblastoma cell lines

目的探讨甘草查尔酮A(LCA)对人胶质瘤U87、U251细胞的增殖抑制和促凋亡作用及其机制研究。方法通过四甲基偶氮唑蓝(MTT)比色法检测细胞增殖抑制作用,并计算出半数抑制浓度(IC50),按IC50和100μM不同浓度进行分组。采用倒置显微镜观察细胞形态,流式细胞术检测细胞凋亡以及Western Blot从蛋白水平检测凋亡相关分子的改变情况。结果 MTT显示LCA对人胶质瘤U87、U251细胞具有明显的增殖抑制作用,呈浓度及时间依赖性,LCA对U87、U251细胞48 h的IC50值分别为61.54μM和53.02μM,倒置显微镜下观察细胞密度减少,形态发生明显改变。流式细胞术结果显示,LCA可促进胶质瘤U87、U251的细胞凋亡(P<0.01)。Western Blot结果显示,LCA干预后可显著降低胶质瘤U87、U251细胞内Bcl-2的表达(P<0.01),相反,增加Bax的表达(P<0.05),呈浓度依赖性。结论 LCA对胶质瘤U87、U251细胞具有明显的增殖抑制作用,可能通过内源性凋亡途径诱导其凋亡。

AbstractObjectiveThe goal of this study is to investigate the cytotoxic effects of licochalcone A on the human glioblastoma cell proliferation and apoptosis and to identify the underlying molecular mechanism.MethodsThe growthinhibiting effect of licochalcone A（LCA） in the human glioblastoma cells were detected by methyl thiazolyl tetrazolium （MTT） assay and calculated the half maximal inhibitory concentration （IC50）, according to different concentrations grouping IC50 or 100 μM. The cell morphological changes were observed under inverted microscope, the cell apoptosis was examined by the flow cytometry and the protein levels of apoptosisrelated molecules were detected by estern Blot.ResultsThe results of MTT revealed that LCA could significantly inhibit U87 and U251 glioma cells proliferation in a dose and timedependent manner. The inhibitory concentrations of LCA for U87 and U251 glioma cells at 48 h were 61.54 μm and 53.02 μm, respectively. The density and morphology of the U87 and U251 glioma cells were remarkably changed under inverted microscope. Results of flow cytometry showed that LCA could induce apoptosis in U87 and U251 glioma cells （P〈0.01）. Furthermore, LCA significantly reduced the level of Bcell lymphoma2（Bcl2） （P〈0.01）, while increased the levels of Bcl2Associated X（Bax） （P〈0.05） in a dosedependent manner.ConclusionLCA could significantly inhibit the cell proliferation of the human glioblastoma cells, subsequently triggering the mitochondrial apoptotic pathway.