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 IC(50) 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 IC(50) 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.
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IC(50)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IC(50)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。.
Keywords: Artesunate; Bcl-2 family proteins; Bortezomib; Leukemia, myeloid, acute; MV4-11 cell lines.