斑蝥酸钠维生素 B6诱导胶质母细胞瘤凋亡研究

The study on apoptosis induced by disodium cantharidinate and vitamin B6 in glioblastoma cells U87

目的：研究斑蝥酸钠维生素B6（ Sodium cantharidinate ，SCA）体外诱导胶质母细胞瘤U87凋亡作用及其机制。方法利用 MTT 法检测0．3125μg/mL，0．625μg/mL，1．25μg/mL，2．5μg/mL，5μg/mL SCA在24 h、48 h和72 h对U87的生长抑制作用，Hoechst 33258染色，荧光显微镜观察SCA作用24 h后U87细胞的形态学变化，利用流式细胞术分别检测SCA作用12 h，24 h后U87细胞凋亡率及细胞周期阻滞。 RT-PCR分析SCA作用24小时后U87凋亡相关基因Bcl-2、Bax和Caspase-3的表达变化。结果 MTT法结果显示SCA对U87细胞的生长抑制作用随着药物浓度的增加而增强，流式细胞术检测结果显示SCA可将细胞周期阻滞于G2/M期，并且可诱导U87细胞凋亡。 Hoechst 33258染色可见经SCA作用后U87细胞出现凋亡形态：染色质凝集、细胞核固缩、部分核碎裂和凋亡小体的形成。 RT-PCR结果显示：SCA作用后，Caspase-3、Bax表达较对照组明显增高（P＜0．05），Bcl-2表达明显下降（P＜0．05）， p53表达无显著增高（P＞0．05）。结论体外实验研究表明SCA可抑制U87细胞增殖，并诱导其凋亡。

Objective The purpose of this study is to investigate the apoptosis mechanisms of glioblasto-ma cell line U87 induced by sodium cantharidinate （ SCA） in vitro.Methods Growth inhibition of U87 by 0.625μg/mL,1.25μg/mL,2.5μg/mL,5μg/mL SCA at 24 h,48 h,72 h were analyzed by MTT assay respec-tively.Morphological changes of U 87 nuclear were detected by fluorescence microscope .U87 cell apoptosis and cell cycle arrest were detected after SCA treatment for 24 h and 48 h by flow cytometry.The changes of apoptosis-related genes Bcl -2,Bax,Caspase-3 expression were analyzed after 24 h of SCA treatment by RT -PCR as-say.Results MTT assay showed that growth inhibition of U 87 cell induced by SCA was accompanied with the in-creased drug concentration ,Hoechst33258 staining showed the morphology of apoptotic U 87 cells nucleui ,chromo-some condensation ,nuclear condensation ,some nuclear fragmentation and formation of apoptotic bodies .Flow cy-tometry showed that SCA could induce cell cycle arrest at the G 2/M phase,and could induce apoptosis of U87.RT-PCR results showed that after 24 h of SCA treatment caspase -3,bax expression of U87 was significantly higher than the control group（P〈0.05）,bcl-2 expression was significantly decreased （P〈0.05）,and P53 expression was not significantly increased（P〉0.05）.Conclusion Our results demonstrate that SCA can inhibit U87 pro-liferation and induce apoptosis of U 87 .