Involvement of Fas ligand-independent caspase8 activation in AsィイD22ィエD2OィイD23ィエD2-induced apoptosis

Fas 配体独立的 caspase8 激活参与 AsiD22D2OD23D2 诱导的细胞凋亡

• 批准号: 10670940
• 负责人: NAOE Tomoki
• 金额: $ 1.6万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10670940/
• 关键词: leukemia; apoptosis; AsィイD22ィエD2OィイD23ィエD2; caspase 8; 白血病; As_< 2> O_< 3>; caspase-8; 急性前骨髄性白血病; 亜ヒ素酸; レドックス

Arsenic trioxide (AsィイD22ィエD2OィイD23ィエD2)-treatment is effective in acute promyelocytic leukemia (APL) patients with t(15;17). Clinically achievable concentrations of AsィイD22ィエD2OィイD23ィエD2 induces apoptosis of NB4, and APl cell line, and some leukemia /cancer cell lines inn vitro. Here, to study the mechanism of AsィイD22ィエD2OィイD23ィエD2-induced apoptosis, we established as AsィイD22ィエD2OィイD23ィエD2-loe sensitive subline, NB4/As. This cell line had a higher concentration of intracellular glutathione (GSH) than NB4 (96 vs. 32 nmol/mg). Reduction of the GSH level by buthionine sulfoxide (BSO) completely restored the sensitivity to AsィイD22ィエD2OィイD23ィエD2. PML-modification and degradation of PMl-RARα were similarly observed on treatment with AsィイD22ィエD2OィイD23ィエD2 in NB4 and NB4 / As, Suggesting their contribution to apoptosis is small. AsィイD22ィエD2OィイD23ィエD2 induced the activation of caspase 3 as well as a loss of mitocondrial transmembrane potential (ΔΨm) in NB4. IN NB4 / As, they were not observed but restored by pretreatment with BSO. Caspase 8 and Bid were activated by the treatment with AsィイD22ィエD2OィイD23ィエD2 in NB4 but not in NB4 / As. In NB4, an inhibitor of caspase 8 blocked not only the activation of caspase 3 and Bid but also the loss of ΔΨm. In both the cell lines, Fas expressed little and agonistic anti-Fas antibody (CH-11) failed to cause apoptosis. These findings suggest that the treatment with AsィイD22ィエD2OィイD23ィエD2 activate caspase 8 in a Fas-ligand independent manner, and that the AsィイD22ィエD2OィイD23ィエD2-resistance in NB4 / As is induced upstream of BSO, AsィイD22ィエD2OィイD23ィエD2might be applied to therapy of leukemias and cancers which are insensitive to low concentrations of AsィイD22ィエD2OィイD23ィエD2.

三氧化二砷（As D2）治疗急性早幼粒细胞白血病（APL） t患者有效（15;17）。临床上可实现的浓度是ィイD22摊位ィエD2Oィイc15ィエD2诱导NB4细胞凋亡,APl细胞系,一些白血病/癌症细胞系客栈体外。为了研究砷化砷诱导凋亡的机制，我们建立了砷化砷敏感子NB4/As。该细胞系细胞内谷胱甘肽（GSH）浓度高于NB4（96对32 nmol/mg）。丁硫氨酸亚砜（BSO）降低GSH水平完全恢复了对As的敏感性。在NB4和NB4 / As中同样观察到pml修饰和PMl-RARα降解，提示PMl-RARα对细胞凋亡的贡献不大。在NB4中，As D2诱导了caspase 3的激活以及线粒体跨膜电位的丧失（ΔΨm）。在NB4 / As中，未观察到，但经BSO预处理后恢复。Caspase 8和Bid在NB4中被As γ γ γ γ γ激活，而在NB4 / As中未被激活。在NB4中，caspase 8的抑制剂不仅阻断了caspase 3和Bid的激活，还阻断了ΔΨm的丢失。在这两种细胞系中，Fas表达很少，且激动性抗Fas抗体（CH-11）未能引起凋亡。这些发现表明,治疗ィイD22摊位ィエD2Oィイc15ィエD2激活半胱天冬酶8 Fas-ligand独立的方式,并且随着ィイD22摊位ィエD2Oィイc15ィエD2-resistance NB4 /诱导BSO的上游,是ィイD22摊位ィエD2Oィイc15ィエD2might适用于治疗白血病和癌症对低浓度的ィイD22摊位ィエD2Oィイc15ィエD2。

项目成果

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