青蒿琥酯诱导人原代白血病细胞凋亡及其相关基因表达谱的初步研究

Artesunate-induced Apoptosis of Human Primary Leukemic Cells and Associated Gene Expression

目的探索青蒿琥酯治疗人类急性白血病的作用及其分子机理。方法收集人类急性白血病患者骨髓细胞,进行原代培养;MTT法检测青蒿琥酯对急性白血病原代细胞增殖的影响;基因芯片观察青蒿琥酯作用前后相关基因表达变化。结果浓度为50,25,12.5μg/mL的青蒿琥酯分别作用于急性白血病原代细胞,48 h后抑制率依次为56.25%、41.49%、30.08%;72 h后抑制率依次为59.8%、45.07%、33.57%,呈剂量、时间依赖关系;青蒿琥酯对AL原代细胞的半数抑制率浓度(IC50)为18.3μg/mL;经青蒿琥酯作用后,AML-M3和ALL-L2原代细胞基因表达谱分别呈现变化趋势,并与药物剂量呈梯度比例关系。结论青蒿琥酯抗AL原代细胞的作用包括多方面,诱导细胞凋亡是其主要作用,可能的机制包括通过Bcl-2途径、线粒体途径、Tnf/TnfR1及Fas/FasL信号转导途径、Jak/Stat通路等诱导AL细胞凋亡;同时青蒿琥酯还有抑制AL原代细胞的增殖及诱导AL原代细胞向成熟分化的作用。

Objective To investigate the therapeutic effect of artesunate for human acute leukemia,and its molecular mechanism.Methods Human acute leukemia bone marrow cells were collected for the primary culture.MTT assay was used to observe the effect of artesunate on acute leukemia cell proliferation.The changes of associated genes expression in acute leukemia cells were examined with gene microarray chips before and after treatment with artesunate.Results After treatment with artesunate at the concentration of 50,25,12.55 μg/mL on the primary acute leukemic cells for 48 hours,the inhibition rate was 56.25 %,41.49 %,30.08 %,respectively;after treatment for 72 hours,the inhibition rates was 59.8 %,45.07 %,33.57 %,respectively.The effect was in dose-and time-dependent manner.Half inhibiting concentration（IC50） was 18.3 μg/mL for artesunate on the primary acute leukemic cells.After treatment with artesunate,the changes of AML-M3 and ALL-L2 gene expression in primary leukemic cells were presented in dose-dependant manner.Conclusion Artesunate exerts multi-target actions on primary acute leukemic cells,and inducing apoptosis is one of the main actions.The possible mechanisms of artesunate in inducing apoptosis of primary acute leukemic cells may be through the BCL-2 pathway,mitochondrial pathway,Tnf/TnfR1 and Fas/FasL signal transduction pathway and Jak/Stat pathways.And the mechanism is also associated with the inhibition of primary leukemic cells proliferation and with the induction of leukemic cells differentiation.