FoxM1抑制剂硫链丝菌素对髓母细胞瘤Daoy细胞增殖与凋亡的影响  

Effect of FoxM1 inhibitor thiostrepton on proliferation and apoptosis of Daoy cells in medulloblastoma

作　　者：郑宜鹏[1] 陈昆[1] 胡远军[1] 林佳平[1] 

机构地区：[1]中山大学附属第一医院神经外科,广州510080

出　　处：《中华神经医学杂志》2013年第5期454-458,共5页Chinese Journal of Neuromedicine

基　　金：广东省科技计划项目（20118080701076）

摘　　要：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、72h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48h后细胞FxoM1mRNA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、72h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0μmol／L硫链丝菌素组比较，1、1．5、2μmol／L硫链丝菌素组细胞FoxM1mRNA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。Objective To investigate the effect of FoxM 1 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.

关 键 词：硫链丝菌素 FOXM1 髓母细胞瘤 细胞增殖 

分 类 号：R734.2[医药卫生—肿瘤]