新生霉素衍生物FM-Nov17抑制慢粒白血病细胞增殖与其诱导DNA损伤的关系  

A Novobiocin Derivate FM-Nov17 Anti-proliferation of CML Cells by Inducing DNA Damage

作　　者：吴丽贤[1] 黄立森 陈显凌[2] 柯方[3] 郑鸣[4] 许建华[1] 

机构地区：[1]福建医科大学药学院药理系,福建省天然药物药理学重点实验室,福州350108 [2]福建医科大学附属协和医院血液科,福建省血液病研究所,福州350001 [3]福建医科大学药学院药物化学系,福州350108 [4]福建医科大学基础医学院人体解剖与胚胎学系,福州350108

出　　处：《福建医科大学学报》2014年第1期1-6,共6页Journal of Fujian Medical University

基　　金：国家自然科学基金(30901824;81173096);福建省自然科学基金杰出青年项目(2011J06013);福建省高校跨世纪优秀人才项目(JA11101)

摘　　要：目的研究新生霉素衍生物FM-Nov17抑制K562和K562/G01细胞增殖的作用与其诱导DNA损伤的关系。方法 3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)及羟基荧光素二醋酸盐琥珀酰亚胺脂(CFSE)染色法检测FM-Nov17对K562及K562/G01细胞增殖的影响;流式细胞光度术检测FM-Nov17诱导K562及K562/G01细胞DNA损伤、细胞周期阻滞和细胞凋亡;Western blot探讨FM-Nov17对DNA损伤、细胞周期和细胞凋亡调控通路相关蛋白表达的影响。结果 FM-Nov17在体外可明显抑制K562和K562/G01细胞增殖,半数抑制率(IC50)分别为(58.28±0.31)和(62.36±0.14)μmol/L,并减少K562及K562/G01细胞的增殖分裂代数;FM-Nov17能诱导细胞DNA损伤和阻滞细胞于G2/M期,并增加细胞凋亡率,呈剂量依赖关系;FM-Nov17能增加γ-H2AX、ATM、P53的磷酸化水平以及Parp和Caspase 3的切割,减少CDC25A和CDC25C的表达。结论FM-Nov17通过诱导DNA的损伤,阻滞细胞于G2/M期,激活线粒体凋亡通路,诱导细胞的凋亡,抑制K562和K562/G01细胞的增殖。Objective To investigate the cytotoxicity effect of FM-Nov17 on K562 and K562/G01 cells in vitro. Methods MTT and CFSE were applied to measure the proliferation inhibition ratio of K562 and K562/G01 cells; Flow Cytometry （FCM） was used to test the level of DNA damage, cell cycle progression, mitochondria membrane potential （MMP） and apoptosis ratio; the level of proteins expression was tested by Western blot. Results FM-Nov17 could significantly inhibited the proliferation of K562 and K562/G01 cells with IC50 was （58.28±0.31）and （62.36±0.14）μmol/L, respectively. FM- Nov17 could induce DNA damage obviously and activate ATM-p53-7 H2AX pathway and checkpoint-related kinases Chkl/Chk2, subsequently led to cell cycle S phase arrest. Furthermore, FM-Nov17 induced apoptotic cell death through mitochondrial membrane potential decrease and activating caspase-3 and Parp cleavage. Conclusion FM-Nov17 has proliferation inhibition potential on K562 and K562/G01 cells through inducing DNA damage and causing cell cycle arresting, resulting in increasing apoptosis ratio in CML cells.

关 键 词：新生霉素 慢性粒细胞白血病 BCR—ABL 凋亡 细胞周期 DNA损伤 

分 类 号：R733.72[医药卫生—肿瘤]