Liu Jing-Jing, Fei Ai-Mei, Nie Rui-Min, Wang Jin, Li Ying, Wang Zhen-Yi, Mi Jian-Qing
State Key Laboratory of Medical Genomics, Shanghai Institute of Hematology, Department of Hematology, Ruijin Hospital Affiliated to Shanghai Jiaotong University School of Medicine, Shanghai 200025, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2011 Jun;19(3):607-11.
The aim of this study was to investigate the apoptosis-inducing effect of artemisinin derivative SM1044 on Kasumi-1 cells and its possible mechanism. Kasumi-1 cells were treated with different concentrations of SM1044, the cell viability was evaluated by MTT assay. Cell apoptosis and cell cycle progression were assessed by using flow cytometry with Annexin-V/PI double staining and flow cytometry with PI staining respectively. The expression of apoptosis-related proteins caspase 3, PARP and the fusion protein AML1-ETO were detected by Western blot. The results indicated that SM1044 inhibited cell growth of Kasumi-1 cells in time- and dose-dependent manners. After exposure of Kasumi-1 cells to 1 µmol/L SM1044 for 24 hours, the cell viability was decreased to 50%. IC(50) of SM1044 to Kasumi-1 cells at 48 hours was 0.17 ± 0.067 µmol/L. SM1044 induced cell apoptosis in a caspase-dependent manner, and the apoptotic rate of Kasumi-1 cells increased as SM1044 concentration increased. Flow cytometry with PI staining revealed that SM1044 induced cell cycle arrest, and the proportion of cells in G(0)/G(1) phase increased from 58.33 ± 4.46% to 71.75 ± 2.24% after exposure to 5 µmol/L SM1044 for 24 hours. Western blot showed that SM1044 increased the expression of apoptosis-related proteins cPARP and cleaved caspase 3 and also degraded the AML1-ETO fusion protein. It is concluded that SM1044 can inhibit the proliferation of Kasumi-1 cells, induce cell apoptosis which may be related to the increased level of cleaved PARP and cleaved caspase 3. SM1044 can also induce cell arrest in G(0)/G(1) phase. As the fusion protein AML1-ETO degrades obviously, it can be the potential target of SM1044 in Kasumi-1 cells.
本研究旨在探讨青蒿素衍生物SM1044对Kasumi-1细胞的凋亡诱导作用及其可能机制。用不同浓度的SM1044处理Kasumi-1细胞，采用MTT法评估细胞活力。分别采用Annexin-V/PI双染流式细胞术和PI染色流式细胞术评估细胞凋亡和细胞周期进程。通过蛋白质免疫印迹法检测凋亡相关蛋白caspase 3、PARP以及融合蛋白AML1-ETO的表达。结果表明，SM1044以时间和剂量依赖性方式抑制Kasumi-1细胞的生长。Kasumi-1细胞暴露于1 μmol/L SM1044 24小时后，细胞活力降至50%。SM1044作用48小时对Kasumi-1细胞的半数抑制浓度（IC50）为0.17±0.067 μmol/L。SM1044以caspase依赖性方式诱导细胞凋亡，且随着SM1044浓度增加，Kasumi-1细胞的凋亡率升高。PI染色流式细胞术显示，SM1044诱导细胞周期阻滞，暴露于5 μmol/L SM1044 24小时后，G(0)/G(1)期细胞比例从58.33±4.46%增加到71.75±2.24%。蛋白质免疫印迹法显示，SM1044增加了凋亡相关蛋白cPARP和裂解的caspase 3的表达，同时降解了AML1-ETO融合蛋白。结论是，SM1044可抑制Kasumi-1细胞增殖，诱导细胞凋亡，这可能与裂解的PARP和裂解的caspase 3水平升高有关。SM1044还可诱导细胞阻滞于G(0)/G(1)期。由于融合蛋白AML1-ETO明显降解，它可能是SM1044在Kasumi-1细胞中的潜在靶点。
