新型化合物XN4抑制急性粒细胞白血病细胞增殖与其诱导氧化性的DNA损伤的关系  

XN4 inhibits proliferation of AML cells by inducing oxidative DNA damage

作　　者：吴丽贤[1,2] 黄立森 陈显凌[3,4] 柯方[5] 郑鸣[6] 许建华[1,2] 

机构地区：[1]福建医科大学药学院药理系,福建福州350004 [2]福建省天然药物药理学重点实验室,福建福州350004 [3]福建医科大学附属协和医院血液科,福建福州350001 [4]福建省血液病研究所,福建福州350001 [5]福建医科大学药学院药化系,福建福州350108 [6]福建医科大学人体解剖与组织胚胎学系,福建福州350108

出　　处：《中国药理学通报》2014年第4期478-483,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 30901824;81173096);福建省自然科学基金杰出青年项目(No 2011J06013);福建省高校跨世纪优秀人才项目(No JA11101)

摘　　要：目的研究XN4在体外抑制急性粒细胞白血病细胞(AML)增殖的作用与诱导DNA损伤的关系。方法 MTT法检测XN4对AML细胞增殖抑制作用;流式细胞术检测AML细胞反应性氧自由基(ROS)、DNA损伤、细胞周期和细胞凋亡;Western blot探讨XN4对相关蛋白表达的影响。结果 XN4明显抑制AML细胞增殖,半数抑制率(IC50)分别为(2.79±0.15)μmol·L-1和(2.76±0.20)μmol·L-1;XN4增加细胞ROS水平和r-H2AX的表达,诱导细胞阻滞在S期并增加细胞凋亡率;XN4能增加H2AX、ATM的磷酸化以及Parp和Caspase-3的切割,而减少CDK2和Cyclin E1的表达。结论 XN4通过激活ROS,诱导DNA的损伤和细胞周期S期阻滞,抑制DNA损伤修复和诱导细胞的凋亡,抑制HL-60及KG1α细胞的增殖,抗氧化剂NAC(N-乙酰半胱氨酸)能减少ROS的产生逆转XN4的作用。Aim To investigate the cytotoxicity of XN4 against AML cells, and the underlying mechanisms by which XN4 might induce DNA damage and apoptotic cell death through reactive oxygen species （ ROS ） . Methods The proliferation inhibition ratio of AML cells was measured by MTT. The level of extracellular ROS, DNA damage, cell cycle process and apoptosis were tested by flow cytometry （ FCM ） . Western blot was applied to test the expression of proteins. Results XN4 significantly inhibited the proliferation of HL-60 and KG1α with IC50 （ 2. 79 ＆#177; 0. 15 ） μmol ＆#183; L-1 and （2. 76 ＆#177; 0. 20） μmol＆#183;L-1 respectively. XN4 signifi-cantly increased the generation of intracellular ROS, followed by inducing DNA damage and activating the ATM-γ-H2AX signaling, which led to increases of cells in the S phases of the cell cycle. Subsequently, XN4 induced apoptotic cell death through activation of caspase-3 and Parp. Moreover, the above effects were all reversed by the ROS scavenger N-acetylcysteine （ NAC ） . Conclusion XN4-induced DNA damage and cell apoptosis in AML cells are mediated via ROS generation.

关 键 词：急性粒细胞白血病 DNA损伤 细胞周期阻滞 细胞凋亡 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]