Induction of mitotic arrest and apoptosis by evodiamine in human leukemic T-lymphocytes
Induction of mitotic arrest and apoptosis by evodiamine in human leukemic T-lymphocytes
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DOI：
10.1016/j.lfs.2003.11.025
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发表时间：
2004-05-21
期刊：
影响因子：
6.1
通讯作者：
Teng, CM
中科院分区：
文献类型：
--
作者：
Huang, YC;Guh, JH;Teng, CM
白血病是一组异质性疾病，其特征在于造血系统异常血细胞的不受控制的增殖。吴茱萸碱是从吴茱萸果实中提取的一种特征性生物碱，对多种肿瘤细胞的增殖和迁移有抑制作用。然而，目前还没有报道阐明这种潜在的天然化合物的作用靶点和抗癌机制。在这项研究中，我们已经确定了吴茱萸碱在人急性白血病CCRF-CEM细胞中的抗增殖和凋亡机制。根据MTT分析，吴茱萸碱以浓度依赖性方式抑制细胞活力，IC 50为0.57 +/- 0.05 μ M。流式细胞仪分析显示，吴茱萸碱诱导的细胞凋亡过程中，细胞周期阻滞于G2/M期。利用瑞氏-姬姆萨染色，我们观察到吴茱萸碱引起细胞有丝分裂停滞。它还深刻地引起这些细胞中聚合微管蛋白水平和丝氨酸70上Bcl-2磷酸化的增加。这些数据表明，微管细胞骨架似乎是响应吴茱萸碱的细胞靶点之一。此外，与吴茱萸碱的CCRF-CEM细胞的治疗与促凋亡蛋白Bax的水平增加，caspase-3的激活，和聚（ADP-核糖）聚合酶，内源性caspase-3底物的蛋白水解裂解。两者合计，我们表明，吴茱萸碱导致有丝分裂停滞和随后的细胞凋亡的CCRF-CEM细胞通过提高聚合微管蛋白水平。此外，一些生物学事件，包括Bcl-2磷酸化，Bax的上调和caspase-3活性的增加可以解释吴茱萸碱诱导的细胞凋亡。(C)2004爱思唯尔公司All rights reserved.
Leukemias are a heterogeneous group of diseases characterized by uncontrolled proliferation of abnormal blood cells of hematopoietic system. Evodiamine, a characteristic alkaloid extracted from Evodia fruits, has been reported to exhibit inhibitory effect on cell proliferation and migration in several types of cancer cells. However, there is no report elucidating the action target and anti-cancer mechanism of this potential natural compound. In this study, we have defined the anti-proliferative and apoptotic mechanisms of evodiamine in human acute leukemia CCRF-CEM cells. According to the MTT assay, the cell viability was inhibited by evodiamine in a concentration-dependent manner with an IC50 Of 0.57 +/- 0.05 muM. Flow cytometry analysis showed that the apoptotic cell death proceeded by evodiamine was accompanied with a cell cycle arrest at the G2/M phase. Using Wright-Giemsa staining, we observed that evodiamine caused the cells to arrest in mitosis. It also profoundly caused an increase in polymerized tubulin levels and Bcl-2 phosphorylation on serine 70 in these cells. These data imply that the microtubular cytoskeleton appears to be one of the cellular targets in response to evodiamine. Moreover, treatment of CCRF-CEM cells with evodiamine was associated with increased levels of pro-apoptotic protein Bax, activation of caspase-3, and proteolytic cleavage of poly (ADP-ribose) polymerase, an endogenous caspase-3 substrate. Taken together, we demonstrate that evodiamine causes the mitotic arrest and a consequent apoptosis in CCRF-CEM cells through the enhancement of polymerized tubulin levels. Furthermore, several biological events including the Bcl-2 phosphorylation, Bax up-regulation and increase of caspase-3 activity could explain evodiamine-induced cell apoptosis. (C) 2004 Elsevier Inc. All rights reserved.