青蒿琥酯对白血病细胞增殖和凋亡的影响

The effects of cell proliferation and apoptosis induced by Artesunate on leukemia cell

目的研究青蒿琥酯对髓系白血病细胞株K562的增殖抑制和凋亡作用,分析其抗肿瘤机制。方法体外培养K562细胞应用不同浓度的青蒿琥酯处理细胞,台盼蓝染色分析细胞活力,WST-1还原法检测细胞增殖,流式细胞术分析细胞凋亡和周期,采用免疫印迹技术分析Bax和Bcl-2表达。结果 K562细胞活力随着药物浓度的增加而下降,而FTY720对细胞增殖的抑制作用随药物浓度增加而增加。药物作用72 h,K562细胞的IC50值为95μmol/L;与对照组比较,50μmol/L和100μmol/L青蒿琥酯处理48 h导致S期细胞减少,G_2M期细胞增加;当浓度达到200μmol/L后,G_2M期细胞减少,但死亡细胞增加到39.65%。双染和流式细胞术分析发现,100μmol/L青蒿琥酯作用24 h后,早期凋亡细胞百分率增加;与对照组比较青蒿琥酯导致细胞内Bax表达增加。结论青蒿琥酯可通过诱导细胞凋亡和抑制细胞增殖而发挥抗肿瘤作用,其线粒体途径可能参与细胞凋亡过程。

Objective To study the effect of artesunate on inhibition of proliferation and apoptosis of myeloid leukemia cell line K562,and analyze its mechanism of antitumor.Methods In vitro,K562 cells were cultivated and applicated by different concentrations of artesunate.The cell vitality was analyzed by trypan blue staining.Proliferation was analyzed by WST-1 reduction method.Apoptosis and cycle of cells were analyzed by flow cytometry,Bax and Bcl-2 expressions were tested by Western blotting.Results K562 cell vitality decreased with the increase of drug concentration,and inhibition of cell proliferation was increased with the increase of FTY720 drug concentration.After 72 h of drug effect,K562 IC50 was 95 μmol/L.Compared with those of control group,50 and 100 μmol/L artesunate for 48 h caused a decline of K562 cells in the S phase,and slightly increased in the G_2M phase.Artesunate(200 μmol/L) induced reduction of K562 cells obviously in G_2M phase,and the death rate was 39.65%.At 24 h,early apoptotic cells rate was increased in the concentration of100 μmol/L artesunate by double staining analysis and flow cytometry.Compared with that of control group,Bax expression was higher by artesunate.Conclusions Artesunate plays a role on anti-tumor by inducing cell apoptosis and inhibiting cell proliferation.The mitochondrial pathway may be involved in the process of cell apoptosis.