双氢青蒿素与三氧化二砷对急性髓系白血病细胞凋亡的影响

Effect of Dihydroartemisinin and Arsenic Trioxide on Apoptosis of Acute Myeloid Leukemia Cells

目的:观察双氢青蒿素(DHA)联合三氧化二砷(ATO)对急性髓系白血病(AML)FLT3-ITD突变MOLM13细胞活力和凋亡的影响及其机制。方法:采用DHA和ATO单药或联合作用于MOLM13细胞,CCK-8法检测细胞活力,细胞平板克隆形成实验检测细胞增殖能力,流式细胞术检测细胞凋亡及活性氧(ROS)产生,Western blot检测凋亡相关蛋白的表达水平。结果:与对照组相比,DHA和ATO单药或联合处理均可抑制细胞生长,激活ROS产生,从而诱导细胞凋亡,而DHA和ATO之间具有协同作用。Western blot结果显示,DHA能促进ATO通过降低Mcl-1、FLT3-ITD表达,显著上调c-PARP表达水平,激活细胞凋亡。结论:DHA联合ATO通过抑制Mcl-1表达、诱导FLT3-ITD蛋白降解诱导FLT3-ITD AML细胞株MOLM13发生凋亡。

Objective: To investigate the effect of dihydroartemisinin(DHA) combined with arsenic trioxide(ATO) on the viability and apoptosis of acute myeloid leukemia(AML) FLT3-ITD mutant cell line MOLM13 and its mechanism. Methods: MOLM13 cells were treated with DHA or ATO alone or in combination. The viability of MOLM13 cells was detected by CCK-8 assay, cell proliferation was observed by colony formation assay, cell apoptosis and reactive oxygen species(ROS) level were measured by flow cytometry, and the expression levels of proteins related to apoptosis were detected by Western blot. Results: Compared with the control group, treatment with DHA and ATO alone or in combination could inhibit cell proliferation, activate ROS formation, and finally induce cell apoptosis. DHA in combination with ATO produced a synergistic effect. Western blot analysis showed that DHA combined with ATO could significantly upregulate the level of c-PARP and activate apoptosis via inhibition of Mcl-1 and FLT3-ITD.Conclusion: DHA combined with ATO induces the apoptosis of FLT3-ITD AML cell line MOLM13 by inhibiting Mcl-1pathway and activating FLT3-ITD protein degradation.