香加皮宝霍甙-Ⅰ诱导人食管癌细胞Eca-109凋亡的实验研究

Effect of Baohuoside-Ⅰon Apoptosis of Human Esophageal Carcinoma Cell Eca-109

背景与目的:研究香加皮宝霍甙-Ⅰ诱导人食管癌细胞Eca_109的凋亡作用及其作用机制。材料与方法:采用MTT法分析不同浓度(12.5、25、50μg/ml)宝霍甙-Ⅰ分别作用Eca_109细胞24h、48h、72h后,对细胞增殖的抑制作用;经不同浓度(12.5、25、50μg/ml)宝霍甙-Ⅰ作用Eca-109细胞48h后,用流式细胞术分析细胞凋亡率及凋亡相关蛋白Survivin的表达;用透射电镜观察凋亡细胞的超微结构变化;用RT-PCR技术检测SurvivinmRNA的表达。结果:不同浓度宝霍甙-Ⅰ均可明显抑制Eca-109细胞的增殖(P均<0.05)且随浓度的增加和作用时间的延长抑制作用增强,作用48h后的半数抑制浓度IC50为24.8μg/ml。不同浓度宝霍甙-Ⅰ作用48h后,均可诱导Eca_109细胞凋亡,50μg/ml时细胞凋亡率达55.26,且导致Eca-109细胞发生凋亡特征性超微结构改变,并使Eca-109细胞SurvivinmRNA和蛋白表达水平均明显降低(P<0.01)。结论:香加皮宝霍甙-Ⅰ可抑制Eca-109细胞增殖,诱导细胞凋亡,该作用可能与下调细胞Survivin表达有关。

BACKGROUND AND AIM： Effect of baohuoside-Ⅰ from Cortex Periplocae on apoptosis of human esophageal carcinoma cell Eca-109 and its mechanism were studied. MATERIALS AND METHODS： After treatment with baohuoside-Ⅰ at different concentrations （12.5, 25, 50 μg/ml）for 24 h, 48 h, 72 h, the inhibitory effect on proliferation of Eca-109 cells was analyzed by MTT method.After treatment with baohuoside-Ⅰ under different concentrations（12.5,25,50 μg/ml） for 48 h, cell apoptotic ratio and expression of Eca-109 cells Survivin protein of were measured with flow eytometry （FCM）; the uhrastructure was examined by transmission electron microscope; the expression of Survivin mRNA was detected by RT-PCR. RESULTS： Baohuoside-Ⅰ significantly inhibited proliferation of Eca-109 cells, and in concentration- dependent manner（P all〈0.05）, the IC50 was 24.8 μg/ml. After treatment with different concentrations of Baohuoside-Ⅰ for 48 h, cell apoptosis of Eca-109 cells was induced significantly （the apoptotic ratio is 55.26% treated with 50 μg/ml of baohuoside-Ⅰ and showing characteristic uhrastructural changes of apoptosis. Expression levels of Survivin mRNA and protein were decreased significantly（P 〈 0.01）. CONCLUSION： Baohuoside-Ⅰ of Cortex Periplecae could inhibit the proliferation and induce apoptosis of Eca-109 cell. This effect was associated with down-regulation of Survivin mRNA expression.