cMyc抑制剂10058-F4诱导白血病THP1细胞凋亡和DNA损伤的机制  

Mechanism of cMyc inhibitor 10058-F4 inducing apoptosis and DNA damage in leukemia THP1 cells

作　　者：魏宇靖[1,2] 潘婕 柯波[1,2] 符环[1,2] 万才水 丁伟荣[1,2] 程洪波 WEI Yujing;PAN Jie;KE Bo;FU Huan;WAN Caishui;DING Weirong;CHENG Hongbo(不详;Department of Hematology,Jiangxi Provincial People′s Hospital,Nanchang 330006,China;Jiangxi Key Laboratory of Hematological Oncology and Cell Biology,Jiangxi Provincial People′s Hospital,Nanchang 330006,China)

机构地区：[1]江西省人民医院血液内科,南昌330006 [2]江西省血液肿瘤细胞生物学重点实验室,南昌330006

出　　处：《实用医学杂志》2022年第3期318-323,329,共7页The Journal of Practical Medicine

基　　金：江西省卫生计生委科技计划(编号:20203020)。

摘　　要：目的研究cMyc抑制剂10058-F4对白血病THP1细胞的抑制作用及分子机制。方法体外培养人白血病THP1细胞,加入不同浓度(1、5、10、50、100μmol/L)的cMyc抑制剂10058-F4处理THP1细胞,CCK-8分析药物对细胞生长抑制作用;Calcein/PI和JC-1荧光染色分别检测细胞凋亡和线粒体膜电位的变化;流式细胞仪检测细胞周期;Western blot分析凋亡相关蛋白(Cleaved-Caspase-3、Bax)和DNA损伤应答相关蛋白(pATM和γH2AX)表达。结果不同浓度10058-F4处理THP1细胞后明显抑制细胞的增殖,并呈浓度依赖和时间依赖性;10058-F4呈浓度依赖地诱导THP1细胞凋亡,并且降低线粒体膜电位水平,差异具有统计学意义(P<0.05);10058-F4可诱导THP1细胞发生G2/M期阻滞,上调促凋亡蛋白(Cleaved-Caspase-3和Bax)以及DNA损伤应答相关蛋白(pATM和γH2AX)的表达水平,差异具有统计学意义(P<0.05)。结论cMyc抑制剂10058-F4对白血病THP1细胞具有凋亡诱导作用,可能与其诱导细胞DNA损伤并激活相关信号途径有关。Objective To investigate the effect and molecular mechanism of cMyc inhibitor 10058-F4 on leukemia THP1 cells.Methods Human leukemic THP1 cells were cultured in vitro and treated with different concentrations(1,5,10,50,100μmol/L)of cMyc inhibitor 10058-F4.The effect of cell growth inhibition was measured by CCK-8 assays.The changes of apoptosis and mitochondrial membrane potential were measured by Calcein/PI and JC-1 fluorescence staining,respectively.The cell cycle was determined by flow cytometry.The expression levels of apoptosis-related proteins(Cleaved-Caspase-3 and Bax)and DNA damage response-related proteins(pATM andγH2 AX)were analyzed by Western blot.Results Different concentrations of 10058-F4 significantly inhibited THP1 cell proliferation in concentration-dependent and time-dependent manners.10058-F4 induced apoptosis and reduced mitochondrial membrane potential levels in a concentration-dependent way,and the difference was statistically significant(P<0.05).Cell cycle assays showed that 10058-F4 could induce phase G2/M arrests in THP1 cells.Western blot experiments showed that the expression of the apoptotic(Cleaved-Caspase-3 and Bax)and DNA damage related proteins(pATM andγH2 AX)were significantly increased in THP1 cells treated with 10058-F4,and the difference was statistically significant(P<0.05).Conclusions The cMyc inhibitor10058-F4 could induce the apoptosis of leukemic THP1 cells by inducing cellular DNA damage and activating relevant signal pathways.

关 键 词：白血病 细胞凋亡 cMyc抑制剂10058-F4 DNA损伤 

分 类 号：R557.2[医药卫生—血液循环系统疾病]