[As2S2诱导K562细胞凋亡分子机制的初步研究]

[Preliminary study on the molecular mechanism of K562 cell apoptosis induced by As2S2].

作者信息

Li Jun-e, Sun Guan-lin, Wu Ying-li, Wu Wei-li

机构信息

Shanghai Institute of Hematology, Ruijin Hospital, Shanghai Second Medical University, Shanghai 200025, China.

出版信息

Zhonghua Zhong Liu Za Zhi. 2003 May;25(3):220-4.

PMID:12839680

Abstract

OBJECTIVE

To investigate the apoptotic inducing effect of As(2)S(2) on K562 cells.

METHODS

The apoptotic inducing effect of As(2)S(2) on K562 cells was determined 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

Apoptosis of K562 cells was induced by 48 - 72 h exposure to 5 micromol/L As(2)S(2). Apoptosis was induced in (34.4 +/- 3.3)% treated cells by 72 h exposure to 3 micro mol/L As(2)S(2), in (21.8 +/- 3.6)% treated cells by 48 h exposure to 5 micromol/L As(2)S(2) and in (46.0 +/- 5.2)% treated cells by 72 h exposure to As(2)S(2) at the same concentration. With 5 micromol/L As(2)S(2), the protein level of Bcr-Abl and JAK2 decreased, while bax expression was upregulated and c-myc was downregulated both in protein and mRNA level. The activity of caspase 3 in K562 cells was increased by As(2)S(2). As(2)S(2) also induced apoptosis of fresh mononuclear cells derived from chronic myelogenous leukemia (CML) patients.

CONCLUSION

As(2)S(2) can induce apoptosis of CML cells. The decline of Bcr-Abl may play an important role. The upregulation of bax, increase of the activity of caspase 3, downregulation of c-myc and decrease of JAK2 may also be involved in the mechanism.

摘要

目的

研究As₂S₂对K562细胞的凋亡诱导作用。

方法

采用流式细胞术、DNA片段化分析及形态学观察检测As₂S₂对K562细胞的凋亡诱导作用。通过蛋白质免疫印迹法检测蛋白表达。采用逆转录聚合酶链反应（RT-PCR）评估基因表达变化。

结果

5 μmol/L As₂S₂作用48 - 72小时可诱导K562细胞凋亡。3 μmol/L As₂S₂作用72小时，诱导（34.4±3.3）%的处理细胞凋亡；5 μmol/L As₂S₂作用48小时，诱导（21.8±3.6）%的处理细胞凋亡；相同浓度As₂S₂作用72小时，诱导（46.0±5.2）%的处理细胞凋亡。5 μmol/L As₂S₂作用后，Bcr-Abl和JAK2蛋白水平降低，bax表达上调，c-myc蛋白和mRNA水平均下调。As₂S₂可增加K562细胞中caspase 3的活性。As₂S₂还可诱导慢性粒细胞白血病（CML）患者新鲜单个核细胞凋亡。