中华神经外科杂志
中華神經外科雜誌
중화신경외과잡지
Chinese Journal of Neurosurgery
2013年
5期
478-483
,共6页
郭国财%刘佳%薄勇力%兰小磊%姚维成
郭國財%劉佳%薄勇力%蘭小磊%姚維成
곽국재%류가%박용력%란소뢰%요유성
神经胶质母细胞瘤%齐墩果酸%MAPK/ERK信号通路%细胞凋亡%U87 MG细胞
神經膠質母細胞瘤%齊墩果痠%MAPK/ERK信號通路%細胞凋亡%U87 MG細胞
신경효질모세포류%제돈과산%MAPK/ERK신호통로%세포조망%U87 MG세포
Glioblastoma%Oleanolic acid%MAPK/ERK signaling pathway%Apoptosis%U87 MG cell
目的 探讨齐墩果酸(0A)对神经胶质母细胞瘤U87MG细胞增殖及细胞凋亡的影响.方法 以U87MG细胞为研究对象,使用25、50、80、100、150和200μg/ml的OA处理24、48和72h后,采用MTT实验检测OA对体外培养U87MG细胞增殖抑制作用,计算其抑制率；采用划痕法检测OA对U87MG细胞迁移能力的影响；用流式细胞仪分析OA对U87MG细胞周期与凋亡的影响；通过Western blot检测OA对MAPK/ERK信号传导通路活性的影响.结果 50μg/ml以上浓度的OA处理48h后,U87MG细胞的形态发生不同程度的变化,大量细胞脱落.150μg/ml和200μg/ml的OA处理U87MG细胞72h后,生长抑制率可达100％,与对照组相比差异有统计学意义(P＜0.01).25μg/ml的OA处理48 h后,迁移至划痕区的细胞数量相比于对照组明显减少.经50μg/mlOA作用48h后,处于Go/G1期的细胞从64.23％上升至75.03％,说明发生Go/G1期阻滞.同一浓度下,细胞凋亡率达到18.77％.U87MG细胞中MEK和ERK蛋白的磷酸化水平受到明显的抑制.结论 OA可显著抑制U87 MG细胞的生长增殖,呈一定的量效关系；OA可明显抑制U87MG细胞的迁移,并能抑制MAPK/ERK信号传导通路的激活.OA通过抑制MAPK/ERK信号传导通路的激活,可以抑制神经胶质母细胞瘤细胞的增殖生长.
目的 探討齊墩果痠(0A)對神經膠質母細胞瘤U87MG細胞增殖及細胞凋亡的影響.方法 以U87MG細胞為研究對象,使用25、50、80、100、150和200μg/ml的OA處理24、48和72h後,採用MTT實驗檢測OA對體外培養U87MG細胞增殖抑製作用,計算其抑製率；採用劃痕法檢測OA對U87MG細胞遷移能力的影響；用流式細胞儀分析OA對U87MG細胞週期與凋亡的影響；通過Western blot檢測OA對MAPK/ERK信號傳導通路活性的影響.結果 50μg/ml以上濃度的OA處理48h後,U87MG細胞的形態髮生不同程度的變化,大量細胞脫落.150μg/ml和200μg/ml的OA處理U87MG細胞72h後,生長抑製率可達100％,與對照組相比差異有統計學意義(P＜0.01).25μg/ml的OA處理48 h後,遷移至劃痕區的細胞數量相比于對照組明顯減少.經50μg/mlOA作用48h後,處于Go/G1期的細胞從64.23％上升至75.03％,說明髮生Go/G1期阻滯.同一濃度下,細胞凋亡率達到18.77％.U87MG細胞中MEK和ERK蛋白的燐痠化水平受到明顯的抑製.結論 OA可顯著抑製U87 MG細胞的生長增殖,呈一定的量效關繫；OA可明顯抑製U87MG細胞的遷移,併能抑製MAPK/ERK信號傳導通路的激活.OA通過抑製MAPK/ERK信號傳導通路的激活,可以抑製神經膠質母細胞瘤細胞的增殖生長.
목적 탐토제돈과산(0A)대신경효질모세포류U87MG세포증식급세포조망적영향.방법 이U87MG세포위연구대상,사용25、50、80、100、150화200μg/ml적OA처리24、48화72h후,채용MTT실험검측OA대체외배양U87MG세포증식억제작용,계산기억제솔；채용화흔법검측OA대U87MG세포천이능력적영향；용류식세포의분석OA대U87MG세포주기여조망적영향；통과Western blot검측OA대MAPK/ERK신호전도통로활성적영향.결과 50μg/ml이상농도적OA처리48h후,U87MG세포적형태발생불동정도적변화,대량세포탈락.150μg/ml화200μg/ml적OA처리U87MG세포72h후,생장억제솔가체100％,여대조조상비차이유통계학의의(P＜0.01).25μg/ml적OA처리48 h후,천이지화흔구적세포수량상비우대조조명현감소.경50μg/mlOA작용48h후,처우Go/G1기적세포종64.23％상승지75.03％,설명발생Go/G1기조체.동일농도하,세포조망솔체도18.77％.U87MG세포중MEK화ERK단백적린산화수평수도명현적억제.결론 OA가현저억제U87 MG세포적생장증식,정일정적량효관계；OA가명현억제U87MG세포적천이,병능억제MAPK/ERK신호전도통로적격활.OA통과억제MAPK/ERK신호전도통로적격활,가이억제신경효질모세포류세포적증식생장.
Objective To investigate the role of oleanolic acid on the proliferation and Suppression of the migration of Malignant Glioma through the Deactivating MAPK/ERK Signaling Pathway in human Glioblastoma cell line U87.Methods The human Glioblastoma cells of U87 were treated with 25μg/ml,50μg/ml,80μg/rml,100μg/ml,150μg/ml,and 200μg/ml of OA for 24h、48h、and 72h.Cell viability of the proliferation was assessed with MTＴ assay.The migration capabilities were observed with the scratch test.Flow cytometry was adopted to analyze cell cycle and apoptosis.Western blotting was used to detect the activation of the MAPK/ERK Signaling Pathway.Results The morphology of U87 MG cells changed with extensive cell detachment,after 48h treatment of 50μg/ml and a higher concentration of OA.Its growth inhibitory rates on U87 MG cells were up to 100％ under 72h treatment on 150μg/ml and 200μg/ml,and compared with control group P ＜0.01.The48h treatment of 25μg/ml OA reduced the amount of cells migrating into the wounding area.The percentage of cells undergoing G0/G1phase rose from 64.23％ to 75.03％ after the 48h treatment of 50μg/ml OA,indicating that G0/G1anest took place.At the same concentration,the apoptotic rate of U87 MG cells was 18.77％.The phosphorylations of MEK and ERK were both suppressed in U87 MG cells.Conclusion OA can significantly inhibit the growth of U87 cell proliferation,a certain quantity effect characteristic; it can obviously inhibit U87 cell migration,and inhibit the MAPK/ERK signaling pathways leading to the activation.Oleanolic acid has the effects of inhibition on U87 cell proliferation and migration capabilities,also it can induct cell apoptosis through deactivating the MAPK/ERK signaling pathway.
