2-甲氧基雌二醇诱导骨髓增生异常综合征MUTZ-1细胞凋亡机制的研究(英文)  

Mechanism of 2-Methoxyestradiol-Induced Apoptosis in Myelodysplastic Syndrome MUTZ-1 Cell Line

作　　者：夏国华[1] 陈宝安[1] 邵泽叶[2] 芦慧霞[2] Dohner Konstanze Dohner Hartmut 

机构地区：[1]东南大学临床医学院附属中大医院血液科,南京210009 [2]东南大学附属中大医院检验中心,南京21009 [3]乌尔姆大学医学院内科Ⅲ区,德国d89081

出　　处：《中国实验血液学杂志》2007年第2期296-301,共6页Journal of Experimental Hematology

基　　金：This study was supported by National Natural Science Foundation(国家自然科学基金资助项目) (No00-R-318)

摘　　要：为了研究2-甲氧基雌二醇(methoxyestradiol,2-ME)诱导骨髓增生异常综合征-难治性贫血伴原始细胞增多型(MDS-RAEB)细胞株MUTZ-1细胞凋亡的机制,将不同浓度的2-甲氧基雌二醇分别与MUTZ-1细胞在体外培养,同时设二甲亚砜和空白对照组。采用四甲基偶氮唑蓝(MTT)比色法测定2-ME对MUTZ-1细胞的生长抑制率,瑞氏-姬姆萨染色后观察2-ME引起细胞的形态学改变,流式细胞术分析细胞周期和凋亡率的变化,贝克曼全自动生化分析仪(synchron clinical system LX20)检测培养上清液中乳酸脱氢酶(lactate dehydrogenase,LD)的变化,DNA凝胶电泳验证2-ME诱导的细胞凋亡。结果表明:2-ME对MUTZ-1细胞的增殖具有明显的抑制作用,该细胞凋亡率明显升高,并呈现时间和剂量依赖性,经统计学处理与对照组相比较有显著性差异(P<0.05)。4μmol/L2-ME作用MUTZ-1细胞12小时后,细胞呈现典型的凋亡细胞形态特征;2-ME作用24小时后MUTZ-1细胞出现G2/M期阻滞;培养上清液中LD含量与对照组相比明显升高,差异具有显著性(P<0.05);4μmol/L2-ME作用MUTZ-1细胞48小时后,DNA凝胶电泳可见明显的DNA梯形条带。结论:2-ME对骨髓增生异常综合征细胞株MUTZ-1有较强的抗肿瘤效应,可能与细胞G2/M期阻滞引起的细胞凋亡有关;2-ME是一种有发展潜力的治疗骨髓异常综合征的药物。The study was aimed to investigate the mechanism of proliferation inhibition and apoptosis of MDS-RAEB MUTZ-1 cells induced by 2-methoxyestradiol （2-ME）, the cell proliferation was determined by MTT assay, apoptosis rate was determined with annexinV-FITC/PI double staining and cell cycle was analyzed by flow cytometry （FCM） after MUTZ-1 cells were treated with different concentrations of 2-ME; the changes of morphologic features of MUTZ-1 cells were observed with Wright-Giemsa~ staining; lactate dehydrogenase was determined by Beckman Counter; and agarose gel electrophoresis was used to verify whether 2-ME can induce apoptosis of MUTZ-1 cells. The results showed that 2- ME inhibited the proliferation of MUTZ-1 cells in a dose-and time-dependent manner and caused a sustained arrest at G2/ M phase in MUTZ-1 cells; the typical apoptotic morphological features appeared in MUTZ-1 cells; the production of lactate dehydrogenaSe was up-regulated and the marked DNA ladder pattern of intemucleosomal fragmentation was observed. It is concluded that the mechanism of proliferation inhibition and apeptosis of MUTZ-1 cells induced by 2-ME is probably related with the G2/M cell cycle arrest; 2-ME may be a potentially adjunctive anticancer drug useful to treat myelodysplastic syndrome.

关 键 词：2一甲氧基雌二醇 骨髓增生异常综合征 MUTZ-1细胞株 细胞凋亡 

分 类 号：R733.7[医药卫生—肿瘤]