高三尖杉酯碱联合三氧化二砷诱导人急性髓系白血病U937细胞株凋亡的实验研究

Combination of Homoharringtonine with Arsenic Trioxide Induces Apoptosis of Human Acute Myeloid Leukemia Cell Line U937

目的:本探讨高三尖杉酯碱(HHT)联合三氧化二砷(As_2O_3)对人急性髓系白血病细胞株U937细胞的增殖抑制和凋亡诱导作用及相关机制。方法:用四甲基偶氮唑(MTT)比色法检测不同浓度HHT及As_2O_3单用及联用对U937细胞增殖的影响;应用Annexin-V-FITC/PI双染流式细胞术检测二者单用或联用对U937细胞的凋亡诱导作用,Western blot方法检测U937细胞内P-Akt^(Ser473)、P-Akt^(Thr308)、BCL-XL、BID、MCL-1与P-MCL-1等蛋白的表达。结果:HHT和As_2O_3均可明显抑制U937细胞的增殖,诱导其凋亡,两药联合可明显增加U937细胞早期凋亡率,且两药联合作用后U937细胞P-Akt^(Ser473)、P-Akt^(Thr308)、MCL-1、P-MCL-1与BCL-XL蛋白表达明显下调,而BID蛋白表达无明显变化。结论:HHT与As_2O_3联合可通过抑制PI3K/Akt信号通路及下游MCL-1蛋白协同抑制U937细胞。

Objective：To investigate the effects and mechanisms of the combination of homoharringtonine（HHT）with arsenic trioxide（As2O3） on human myeloid cell line U937 in vitro.Methods：MTT method was used to determine the antiproliferating effect of different concentrations of HHT,As2O3 and their combination on U937 cells;the flow cytometry with Annexin-V-FITC/PI double staining was used to determine the apoptosis- induced effect of HHT and As2O3 alone or their combination;Western blot method was used to detect the protein expression of P-akt-（Ser473）,P—Akt-（Thr308）,BCL-XL,BID,MCL-1,P-MCL-1 and so on.Results：HHT and As2O3 could significantly inhibit proliferation of U937 cells and induce their apoptosis.The combination of these 2 drugs could significantly enhance the early apoptosis of U937 cells.After combination of these 2 drugs was used,the protein expressions of P-Akt-（Ser473）,P-Akt-（Thr308）,MCL-1,P—MCL-1 and BCL-XL were obviously down-regulated,but the expression of BID protein did not change.Conclusion：The combination treatment of HHT and As2O3 can synergistically inhibit the growth of U937 cells through inhibition of PI3K/Akt signal way and MCL4 protein.