Apoptotic effect of As₂S₂on K562 cells and its mechanism  

As2S2诱导K562细胞凋亡及其机制（英文）

作　　者：李俊娥[1] 邬维礼[1] 王振义[1] 孙关林[1] 

机构地区：[1]上海第二医科大学附属瑞金医院上海血液学研究所

出　　处：《Acta Pharmacologica Sinica》2002年第11期33-38,共页中国药理学报（英文版）

基　　金：Supported in part by Grants from the National Natural Science and Technology Foundation of 9th Quinquennial Program of China(969060117), National Natural Science Foundation of China(No 39870773), and Clyde Wu Foundation of Shanghai Institute of Hematolog

摘　　要：AIM: To investigate the apoptotic effect of As2S2 on K562 cells and its mechanism. METHODS: The effect of As2S2on proliferation of K562 cells was determined by counting the number of cells. Apoptosis was assessed by flow cytometry, DNA fragmentation analysis, and morphology observation. Expression of protein was determined by Western blot. RT-PCR was used to evaluate changes in gene expression. RESULTS: As2S2 greatly inhibited the proliferation and induced apoptosis of K562 cells in a concentration- and time-dependent manner at the concentra-tion range of 1-5 μmol/L during 24-72 h. Viable cells were decreased to approximately 71 % of control at the concentration of 5 μmol/L after 48-h incubation, 31.4% after 72-h incubation, and 45.4 % at 3 μmol/L after 72-h incubation. At 3μmol/L for 72 h, 5 μmol/L for 48 h, and 5 μmol/L for 72 h, the apoptosis rate were 34.4 %, 21.8 %, and 46 % of the treated-cells, respectively. As2S2 decreased the Bcr-Abl fusion protein and protein tyrosine kinase (PTK) activity of c-abl and Bcr-Abl, but it did not change the transcription of bcr-abl assayed. As2S2 also induced apoptosis in fresh mononuclear cells derived from chronic myelogenous leukemia (CML) patients. CML Ph+ leukemia cells were more sensitive to the apoptotic effect of As2S2 than Ph mononuclear cells (P【0.05). CONCLUSION: As2S2 inhibited the proliferation and induced apoptosis in K562 and fresh CML mononuclear cells. The decline of the Bcr-Abl protein and its PTK activity may play an important role in the apoptotic effect of As2S2. As2S2 may be a useful agent for the treatment of CML.目的:探索As₂S₂对K562细胞的作用及其机制.方法:As₂S₂对K562细胞的生长抑制作用用细胞计数法;细胞凋亡的检测用流式细胞分析、基因组DNA电泳、细胞形态学观察等方法;Western-blot方法用于蛋白表达的检测;基因表达的变化用半定量RT-PCR方法.结果:As₂S₂浓度在1-5μmol/L作用24-72 h即可抑制K562细胞生长,大于3μmol/L时可诱导K562细胞凋亡.As₂S₂能降低K562细胞中Bcr-Abl蛋白水平及 c-abl和 Bcr-Abl PTK活性,但不调变bcr-abl基因表达水平.As₂S₂也能诱导慢性粒细胞性白血病（CML）患者单个核细胞凋亡,且Ph⁺单个核细胞比Ph^- 单个核细胞对As₂S₂诱导的凋亡更敏感.结论:As₂S₂可通过降低Bcr-Abl蛋白含量而诱导CML细胞凋亡.As₂S₂可能为治疗CML的有效药物.

关 键 词：ARSENICALS chronic myeloid leukemia APOPTOSIS Bcr-Abl fusion proteins 

分 类 号：R73-3[医药卫生—肿瘤]