中华神经外科杂志
中華神經外科雜誌
중화신경외과잡지
Chinese Journal of Neurosurgery
2012年
3期
291-294
,共4页
边心超%孟繁凯%杨福伟%付双林%金鑫%罗毅男%葛鹏飞
邊心超%孟繁凱%楊福偉%付雙林%金鑫%囉毅男%葛鵬飛
변심초%맹번개%양복위%부쌍림%금흠%라의남%갈붕비
雷公藤红素%C6胶质瘤细胞%细胞凋亡%细胞周期阻滞
雷公籐紅素%C6膠質瘤細胞%細胞凋亡%細胞週期阻滯
뢰공등홍소%C6효질류세포%세포조망%세포주기조체
Celastrol%C6 glioma cell%Apoptosis%Cell cycle arrest
目的 探讨雷公藤红素对体外C6胶质瘤细胞凋亡及细胞周期阻滞的影响及机制.方法 雷公藤红素处理体外培养的C6胶质瘤细胞,MTT法检测细胞增殖,Hochest33342染色荧光显微镜观察以及锇铀铅染色透射电镜观察胶质瘤细胞形态变化,流式细胞术分析细胞凋亡率及细胞周期改变.蛋白印迹分析检测凋亡相关蛋白及细胞周期调控蛋白的表达变化.结果 雷公藤红素对C6胶质瘤细胞的增殖抑制作用呈浓度和时间依赖性；2.56 μmol/L雷公藤红素处理C6胶质瘤细胞24h后,荧光显微镜、透射电镜下均可见凋亡形态学改变.流式细胞术检查显示,凋亡细胞比例由1.32％升高到19.34％；并且,处于G0/G1期的细胞比例由76.42％降低到54.52％,G2/M期细胞比例由9.29％升高到30.50％.蛋白印迹分析显示,雷公藤红素降低了Bcl -2及XIAP蛋白表达,促进了Bax及Caspase -3的表达以及PARP的剪切；雷公藤红素在诱导细胞周期调控蛋白p21、p27及cyclin B1蛋白表达增加的同时抑制了cdk2蛋白的表达.结论 雷公藤红素能够通过调节凋亡相关蛋白及细胞周期相关因子的表达影响C6胶质瘤细胞的凋亡及细胞周期阻滞.
目的 探討雷公籐紅素對體外C6膠質瘤細胞凋亡及細胞週期阻滯的影響及機製.方法 雷公籐紅素處理體外培養的C6膠質瘤細胞,MTT法檢測細胞增殖,Hochest33342染色熒光顯微鏡觀察以及鋨鈾鉛染色透射電鏡觀察膠質瘤細胞形態變化,流式細胞術分析細胞凋亡率及細胞週期改變.蛋白印跡分析檢測凋亡相關蛋白及細胞週期調控蛋白的錶達變化.結果 雷公籐紅素對C6膠質瘤細胞的增殖抑製作用呈濃度和時間依賴性；2.56 μmol/L雷公籐紅素處理C6膠質瘤細胞24h後,熒光顯微鏡、透射電鏡下均可見凋亡形態學改變.流式細胞術檢查顯示,凋亡細胞比例由1.32％升高到19.34％；併且,處于G0/G1期的細胞比例由76.42％降低到54.52％,G2/M期細胞比例由9.29％升高到30.50％.蛋白印跡分析顯示,雷公籐紅素降低瞭Bcl -2及XIAP蛋白錶達,促進瞭Bax及Caspase -3的錶達以及PARP的剪切；雷公籐紅素在誘導細胞週期調控蛋白p21、p27及cyclin B1蛋白錶達增加的同時抑製瞭cdk2蛋白的錶達.結論 雷公籐紅素能夠通過調節凋亡相關蛋白及細胞週期相關因子的錶達影響C6膠質瘤細胞的凋亡及細胞週期阻滯.
목적 탐토뢰공등홍소대체외C6효질류세포조망급세포주기조체적영향급궤제.방법 뢰공등홍소처리체외배양적C6효질류세포,MTT법검측세포증식,Hochest33342염색형광현미경관찰이급철유연염색투사전경관찰효질류세포형태변화,류식세포술분석세포조망솔급세포주기개변.단백인적분석검측조망상관단백급세포주기조공단백적표체변화.결과 뢰공등홍소대C6효질류세포적증식억제작용정농도화시간의뢰성；2.56 μmol/L뢰공등홍소처리C6효질류세포24h후,형광현미경、투사전경하균가견조망형태학개변.류식세포술검사현시,조망세포비례유1.32％승고도19.34％；병차,처우G0/G1기적세포비례유76.42％강저도54.52％,G2/M기세포비례유9.29％승고도30.50％.단백인적분석현시,뢰공등홍소강저료Bcl -2급XIAP단백표체,촉진료Bax급Caspase -3적표체이급PARP적전절；뢰공등홍소재유도세포주기조공단백p21、p27급cyclin B1단백표체증가적동시억제료cdk2단백적표체.결론 뢰공등홍소능구통과조절조망상관단백급세포주기상관인자적표체영향C6효질류세포적조망급세포주기조체.
Objective To investigate the effects of celastrol on the apoptosis and cell cycle arrest of C6 glioma cells and its mechanism.Methods C6 glioma cells were cultured in vitro.MTT was used to assay cellular proliferation.Fluorescence microscope and transmission electron microscope were used to evaluate the morphological alterations of C5 glioma cells after being incubated with 2.56 μmol/L celastrol.Flow cytometry was used to analyze the alterations of apoptosis rate and cell cycle.Results The inhibitory effects of celastrol on C6 glioma cells were dependent on celastrol concentration and incubation time.Typical morphological alterations of apoptosis were observed under photo-microscope,fluorescence microscope and transmission electron microscope respectively.Flow cytometry showed that the apoptosis rate increased from 1.32％ in control group to 19.34％ in celastrol group,and cellular quantity decreased from 76.42％ to 54.52％ at G0/G1 phase but increased from 9.29％ to 30.50％ at G2/M.Western blotting analysis showed that the expression of apoptosis -related proteins Bc1-2 and XIAP were down-regulated,but Bax,Caspase-3 and cleaved PARP were up -regulated.Moreover,the expression of cell cycle regulator p21,p2 and cyclin B1 were increased,but cdk2 was decreased.Conclusion Celastrol could induce C6 glioma cell apoptosis and cell cycle arrest via influencing the expression of apoptosis related proteins and cell cycle regulators.
