阿糖胞苷对人急性髓系白血病细胞活力、凋亡的作用及机制

Effect and mechanism of cytarabine on viability and apoptosis in acute myeloid leukemia cells

目的探讨阿糖胞苷(Ara-C)对人急性髓系白血病(AML)细胞活力、凋亡的作用及机制。方法选取对数生长期人AML细胞株MV4-11,分别用二甲基亚砜(DMSO)及0.5、1.0、1.5μmol/L Ara-C作用24、48和72 h,采用细胞计数试剂盒8(CCK8)和流式细胞仪检测各组细胞不同时间的光密度(OD)值和细胞凋亡情况;取对数生长期MV4-11细胞分为空白组和1.0μmol/L Ara-C组,1.0μmol/L Ara-C组细胞分别处理3、6、12、24及48 h,采用蛋白印迹法检测各组MV4-11细胞中Caspase 3、c-Myc及高迁移率族蛋白1(HMGB1)的蛋白表达;利用cBioportal在线分析工具和R包(survminer)分析c-Myc表达水平与AML患者生存期的相关性。结果与DMSO组相比,0.5、1.0及1.5μmol/L Ara-C组MV4-11细胞不同时间的细胞活力均降低、凋亡细胞比例升高,且对Ara-C均有浓度依赖性(P<0.0001或P<0.01);与DMSO组相比,1.0μmol/L Ara-C组MV4-11细胞经不同时间处理后Caspase 3表达水平均降低,且有时间依赖(P<0.05);与DMSO组相比,1.0μmol/L Ara-C组MV4-11细胞经不同时间后,细胞的HMGB1和c-Myc蛋白表达均降低,差异均有统计学意义(P<0.05);c-Myc高表达与AML患者较差生存期呈正相关((P<0.05)。结论Ara-C可抑制人AML细胞活力和诱导细胞凋亡,其机制可能与HMGB1/c-Myc的下调有关。

Objective To investigate the effects and mechanism of cytarabine(Ara-C)on cell viability,apoptosis in acute myeloid leukemia(AML)cells.Methods The logarithmic growth phase AML cell MV4-11 was selected and treated with DMSO and Ara-C(0.5,1.0,and 1.5μmol/L)respectively for 24,48,and 72 h.Optical density(OD)value and apoptosis of each group were detected by Cell Counting Kit-8(CCK8)and flow cytometry.The logarithmic growth phase human AML cell MV4-11 was divided into DMSO group,1.0μmol/L Ara-C group,the latter was treated for 3,6,12,24,and 48 h respectively.The protein expression of Caspase 3,c-Myc,and high mobility group box(HMGB1)in MV4-11 of each group were detected by Western blot.The cBioportal database and R packages(survminer)were used to analyses the correlation between c-Myc expression and survival of acute myeloid leukemia patients.Result The cell viability of MV4-11 cells were reduced significantly in the 0.5,1.0,and 1.5μmol/L Ara-C groups compared with the DMSO group,while proportion of apoptosis MV4-11 cells increased and showed concentration-dependency on Ara-C(P<0.0001 or P<0.01).Compared with the DMSO group,the Caspase 3 expression in MV4-11 cells were decreased in1.0μmol/L Ara-C group and showed time-dependency(P<0.05).Compared with DMSO group,HMGB1 and c-Myc protein expression all decreased in MV4-11 cells in 1.0μmol/L Ara-C group,differences were statistically significant(P<0.05).c-Myc expression was positively correlated with undesired overall survival time in AML patients(P<0.05).Conclusion Ara-C can inhibit cell viability and induce apoptosis of AML.Such mechanism may be related with downregulating of HMGB1/c-Myc.