CAJ | 학술논문

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。
목적：연구반모산납유생소B6（ Sodium cantharidinate ，SCA）체외유도효질모세포류U87조망작용급기궤제。방법이용 MTT 법검측0．3125μg/mL，0．625μg/mL，1．25μg/mL，2．5μg/mL，5μg/mL SCA재24 h、48 h화72 h대U87적생장억제작용，Hoechst 33258염색，형광현미경관찰SCA작용24 h후U87세포적형태학변화，이용류식세포술분별검측SCA작용12 h，24 h후U87세포조망솔급세포주기조체。 RT-PCR분석SCA작용24소시후U87조망상관기인Bcl-2、Bax화Caspase-3적표체변화。결과 MTT법결과현시SCA대U87세포적생장억제작용수착약물농도적증가이증강，류식세포술검측결과현시SCA가장세포주기조체우G2/M기，병차가유도U87세포조망。 Hoechst 33258염색가견경SCA작용후U87세포출현조망형태：염색질응집、세포핵고축、부분핵쇄렬화조망소체적형성。 RT-PCR결과현시：SCA작용후，Caspase-3、Bax표체교대조조명현증고（P＜0．05），Bcl-2표체명현하강（P＜0．05）， p53표체무현저증고（P＞0．05）。결론체외실험연구표명SCA가억제U87세포증식，병유도기조망。
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 .