青蒿琥酯联合硼替佐米对急性髓系白血病细胞体外凋亡、自噬的影响及其作用机制  

Effects of artesunate combined with bortezomib on apoptosis and autophagy of acute myeloid leukemia cells in vitro and its mechanism

作　　者：胡莲洁[1,3] 姜涛 王甫珏[1] 黄世华 成晓敏[4] 贾永前 Hu Lianjie;Jiang Tao;Wang Fujue;Huang Shihua;Cheng Xiaomin;Jia Yongqian(Department of Hematology, Hematology Laboratory, Western China Hospital, Sichuan University, Chengdu 610041, China;Department of Hematology, The People's Hospital of Sichuan Province, Chengdu 610072, China;Department of Hematology, The Second People's Hospital of Yibin, Yibin 644000, Sichuan Province, China;Department of Hematology, Chengdu Military General Hospital, Chengdu 610083, China)

机构地区：[1]四川大学华西医院血液科、血液病研究室,成都610041 [2]四川省人民医院血液科,成都610072 [3]四川省宜宾市第二人民医院血液科,644000 [4]成都军区总医院血液科,610083

出　　处：《中华血液学杂志》2019年第3期204-208,共5页Chinese Journal of Hematology

摘　　要：目的探讨青蒿琥酯联合硼替佐米对急性髓系白血病(AML)细胞株MV4-11细胞增殖、凋亡及自噬的影响及其作用机制。方法 MTT法检测青蒿琥酯、硼替佐米单用及两药联合对MV4-11细胞的增殖抑制作用,流式细胞术检测细胞凋亡情况,Western blot法检测细胞内Bcl-2家族蛋白(Bcl-2、Mcl-1、Bim、Bax)、cleaved-Caspase-3及自噬相关蛋白LC3B的表达。结果青蒿琥酯呈浓度和时间依赖性抑制MV4-11细胞增殖,作用48 h的IC50为1.44 μg/ml。硼替佐米呈浓度依赖性抑制MV4-11细胞增殖,作用48 h的IC50为8.97 nmol/L。青蒿琥酯(0.75、1.0 μg/ml)与硼替佐米(6、8 nmol/L)联合作用48 h,对MV4-11细胞的增殖抑制率均高于单药组(P值均<0.05),两药相互作用的协同指数(CI)均<1。1.5 μg/ml青蒿琥酯作用MV4-11细胞48 h的凋亡率为(15.27±2.18)%,8 nmol/L硼替佐米作用MV4-11细胞48 h的凋亡率为(19.85±3.23)%,两药联合作用后凋亡率增高至(81.67±5.96)%,显著高于单药组(P值均<0.05)。两药联合作用MV4-11细胞24 h后细胞内cleaved-Caspase-3、促凋亡蛋白Bim及自噬相关蛋白LC3B表达上调,抗凋亡蛋白Bcl-2表达下调。结论青蒿琥酯联合硼替佐米具有协同抑制MV4-11细胞增殖和诱导凋亡及促进自噬的效应,作用机制可能与Bcl-2家族蛋白表达的改变有关。Objective To investigate the effects of artesunate combined with bortezomib on the proliferation, apoptosis and autophagy of human acute myeloid leukemia cell lines MV4-11, and its mechanisms. Methods MTT method was used to determine the anti-proliferation effect of different concentrations of artesunate, bortezomib and their combination on MV4-11 cells. The cell apoptosis were analyzed by flow cytometry. The expression of cleaved-Caspase-3, Bcl-2 family protein (Bcl-2, Mcl-1, Bim, Bax) and autophagy-related protein LC3B were assayed by Western blot. Results Artesunate displayed a proliferation inhibition effect on MV4-11 with dose- and time-dependent manner, the IC50 of artesunate on MV4-11 after 48 hours was 1.44 μg/ml. Bortezomib displayed a proliferation inhibition effect on MV4-11 with dose-dependent manner, the IC50 of bortezomib on MV4-11 after 48 hours was 8.97 nmol/L. The combination of artesunate (0.75, 1.0 μg/ml) and Bortezomib (6, 8 nmol/L) showed higher inhibition on MV4-11 than artesunate or bortezomib alone in the same concentration gradient after 48 hours (P<0.05). The cooperation index of the two drugs were all less than 1. The 48 h apoptotic rate of artesunate (1.5 μg/ml) on MV4-11 was (15.27±2.18)%,(19.85±3.23)% of bortezomib (8 nmol/L),(81.67±5.96)% of combination of the two drugs, significantly higher than the single group (P<0.05). When combination of the two drugs on MV4-11 after 24 hours, the levels of pro-apoptotic protein Bim and the cleaved activation of Caspase-3 and autophagy-related protein LC3B were up-regulated and the anti-apoptotic protein Bcl-2 expressions was down-regulated. Conclusion Combination of artesunate with bortezomib shows a significant synergistic effects on proliferation, apoptosis and autophagy of MV4-11 cell lines, which may be associated with Bcl-2 family proteins expression.

关 键 词：白血病 髓样 急性 青蒿琥酯 硼替佐米 MV4-11细胞 BCL-2家族蛋白 

分 类 号：R733.71[医药卫生—肿瘤]