中华神经医学杂志
中華神經醫學雜誌
중화신경의학잡지
CHINESE JOURNAL OF NEUROMEDICINE
2013年
5期
454-458
,共5页
郑宜鹏%陈昆%胡远军%林佳平
鄭宜鵬%陳昆%鬍遠軍%林佳平
정의붕%진곤%호원군%림가평
硫链丝菌素%FoxM1%髓母细胞瘤%细胞增殖
硫鏈絲菌素%FoxM1%髓母細胞瘤%細胞增殖
류련사균소%FoxM1%수모세포류%세포증식
Thiostrepton%FoxM1%Medulloblastoma%Cell proliferation
目的 探讨FoxM1抑制剂硫链丝菌素对髓母细胞瘤Daoy细胞株增殖及凋亡的影响. 方法 体外常规培养髓母细胞瘤Daoy细胞株,CCK-8法检测0、1、1.5、2、3、5μmol/L硫链丝菌素作用24、48、72 h后细胞存活率的变化；实时定量聚合酶链反应(RT-PCR)、免疫印迹法分别检测0、1、1.5、2μmol/L硫链丝菌素作用48 h后细胞FxoMl mRNA和蛋白的表达,流式细胞仪检测细胞周期和凋亡,免疫印迹法检测凋亡相关蛋白Bcl-2、Bax的表达. 结果 CCK-8检测显示不同浓度硫链丝菌素作用后Doay细胞存活率下降,且呈浓度和时间依赖性,差异有统计学意义(P＜0.05).24、48、72 h的IC50分别为(2.1±0.15)、(1.69±0.11)、(1.39±0.1)μmol/L; RT-PCR和免疫印迹实验显示,与0 μmol/L硫链丝菌素组比较,1、1.5、2μmol/L硫链丝菌素组细胞FoxMl mRNA及蛋白水平下降,G2/M期细胞细胞比例和凋亡率升高,凋亡蛋白bax表达增加,抗凋亡蛋白bcl-2表达减少,且均呈浓度依赖性,差异有统计学意义(P＜0.05). 结论 FoxM1抑制剂硫链丝菌素可明显抑制Daoy细胞增殖,可能与阻滞细胞周期于G2/M期、改变bcl-2/bax表达诱发凋亡有关.
目的 探討FoxM1抑製劑硫鏈絲菌素對髓母細胞瘤Daoy細胞株增殖及凋亡的影響. 方法 體外常規培養髓母細胞瘤Daoy細胞株,CCK-8法檢測0、1、1.5、2、3、5μmol/L硫鏈絲菌素作用24、48、72 h後細胞存活率的變化；實時定量聚閤酶鏈反應(RT-PCR)、免疫印跡法分彆檢測0、1、1.5、2μmol/L硫鏈絲菌素作用48 h後細胞FxoMl mRNA和蛋白的錶達,流式細胞儀檢測細胞週期和凋亡,免疫印跡法檢測凋亡相關蛋白Bcl-2、Bax的錶達. 結果 CCK-8檢測顯示不同濃度硫鏈絲菌素作用後Doay細胞存活率下降,且呈濃度和時間依賴性,差異有統計學意義(P＜0.05).24、48、72 h的IC50分彆為(2.1±0.15)、(1.69±0.11)、(1.39±0.1)μmol/L; RT-PCR和免疫印跡實驗顯示,與0 μmol/L硫鏈絲菌素組比較,1、1.5、2μmol/L硫鏈絲菌素組細胞FoxMl mRNA及蛋白水平下降,G2/M期細胞細胞比例和凋亡率升高,凋亡蛋白bax錶達增加,抗凋亡蛋白bcl-2錶達減少,且均呈濃度依賴性,差異有統計學意義(P＜0.05). 結論 FoxM1抑製劑硫鏈絲菌素可明顯抑製Daoy細胞增殖,可能與阻滯細胞週期于G2/M期、改變bcl-2/bax錶達誘髮凋亡有關.
목적 탐토FoxM1억제제류련사균소대수모세포류Daoy세포주증식급조망적영향. 방법 체외상규배양수모세포류Daoy세포주,CCK-8법검측0、1、1.5、2、3、5μmol/L류련사균소작용24、48、72 h후세포존활솔적변화；실시정량취합매련반응(RT-PCR)、면역인적법분별검측0、1、1.5、2μmol/L류련사균소작용48 h후세포FxoMl mRNA화단백적표체,류식세포의검측세포주기화조망,면역인적법검측조망상관단백Bcl-2、Bax적표체. 결과 CCK-8검측현시불동농도류련사균소작용후Doay세포존활솔하강,차정농도화시간의뢰성,차이유통계학의의(P＜0.05).24、48、72 h적IC50분별위(2.1±0.15)、(1.69±0.11)、(1.39±0.1)μmol/L; RT-PCR화면역인적실험현시,여0 μmol/L류련사균소조비교,1、1.5、2μmol/L류련사균소조세포FoxMl mRNA급단백수평하강,G2/M기세포세포비례화조망솔승고,조망단백bax표체증가,항조망단백bcl-2표체감소,차균정농도의뢰성,차이유통계학의의(P＜0.05). 결론 FoxM1억제제류련사균소가명현억제Daoy세포증식,가능여조체세포주기우G2/M기、개변bcl-2/bax표체유발조망유관.
Objective To investigate the effect of FoxM1 inhibitor thiostrepton on proliferation and apoptosis of Daoy cell line in medulloblastoma.Methods Daoy cells from medulloblastoma were routinely cultured in vitro,and they were treated with thiostrepton at various concentrations (0,1,1.5,2,3 and 5 μmol/L); the mRNA and protein levels of FoxM1 were detected by real-time PCR and Western blotting 48 h after treatment; the proliferation of Daoy cells was investigated using CCK-8 assay 24,48 and 72 h after treatment; flow cytometry was used to observe the changes of cycle and apoptosis of Daoy cells after thiostrepton treatment; the effects of thiostrepton on apoptosis-relating proteins of Bcl-2 and Bax were studied using Western blotting.Results Thiostrepton could obviously inhibit Daoy cell proliferation in time-and dose-dependent manners,with significant difference between each two concentrations (P＜0.05),and IC50 at 24,48 and 72 h after treatment was (2.1±0.15),(1.69±0.11) and (1.39±0.1) μmol/L.As thiostrepton concentration increased,FoxM1 mRNA and protein expressions gradually decreased in Daoy cells; increasing thiostrepton concentration induced Daoy cell cycle being gradually blocked at G2/M phase,decreased Bcl-2 expression,up-regulated Bax expression and significantly increased apoptosis rate,with significant differences between each two concentrations (P＜0.05).Conclusion FoxM1 inhibitor Thiostrepton can obviously inhibit Daoy cell proliferation,which might be related to cells being arrested at G2/M stage and apoptosis being induced by Bcl-2/Bax expression proportion changes.
