Apoptosis, necrosis and cell proliferation-inhibition by cyclosporine A in U937 cells (a human monocytic cell line)
Apoptosis, necrosis and cell proliferation-inhibition by cyclosporine A in U937 cells (a human monocytic cell line)
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DOI：
10.1016/j.phrs.2005.12.007
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发表时间：
2006-03-01
影响因子：
9.3
通讯作者：
Tsushida, T
中科院分区：
文献类型：
--
作者：
Roy, MK;Takenaka, M;Tsushida, T
免疫抑制药物环孢素A(CsA)已用于器官移植和自身免疫性疾病的治疗。然而。该药物会对肾脏、肝脏和神经系统造成不良影响，其特征是受影响区域的细胞丢失。细胞凋亡已被证明在CsA诱导的细胞毒性中起作用。由于线粒体膜的通透性是大多数脊椎动物细胞凋亡的共同标准，在这里，我们评估了CsA是否导致导致细胞细胞毒性的途径中的线粒体功能丧失。我们发现CsA在U937细胞系中引起浓度和时间依赖性的细胞活力丧失。10 mU M CsA处理细胞后，细胞周期G0/G1期阻滞，S和G2/M期细胞数同时减少。在与细胞活力丧失相关的机制研究中，10亩环孢素A处理细胞24小时，导致DNA断裂和Annexin-V阳性细胞增加。CsA还可提高半胱氨酸蛋白酶caspase-3的活性，降低线粒体膜电位，诱导细胞色素c释放到胞浆中。此外，CsA处理增加了亚G0/G1峰的细胞数量，这表明DNA减少，尽管当细胞用广泛的caspase抑制剂预处理时，这种增加没有观察到。在这项研究中，我们还发现，当细胞孵育较长时间时，较高剂量的CsA会诱导LDH释放。综上所述，这些数据表明CsA诱导的细胞死亡模式是剂量和时间依赖的。较低剂量的CsA短期孵育会阻止细胞生长；这种停滞与细胞凋亡的发生重叠，然后在较长时间使用较高剂量的CsA治疗后出现坏死。(C)2006爱思唯尔有限公司。保留所有权利。
The immunosuppressive drug cyclosporine A (CsA) has been used in both organ transplantation and the treatment of autoimmune disorders. However. the drug causes adverse effects in the kidney, liver and nervous system, characterized by cellular loss in the affected area. Apoptosis has been shown to play a role in CsA-induced cytotoxicity. Because permeabilization of the mitochondrial membrane is a common criterion in most apoptotic settings in vertebrate cells, here we evaluated whether CsA causes loss of mitochondrial function in the pathway leading to cellular cytotoxicity. We found that CsA caused a concentration- and time-dependent loss of cell viability in the U937 cell line. Treatment of cells at a dose of 10 mu M CsA resulted in G0/G1 arrest with a concurrent decrease in the number of cells in the S and G2/M phases of the cell cycle. In mechanistic studies related to the loss of viability, treating cells with 10 mu M CsA for 24 h resulted in both DNA fragmentation and an increase of annexin-V-positive cells. CsA treatment also increased activity of the cysteine protease caspase-3, decreased the mitochondrial membrane potential and induced the release of cytochrome c into the cytosol. Furthermore, CsA treatment increased the number of cells in the sub-G0/G1 peak, indicative of a reduction in DNA, although this increase was not observed when cells were pre-treated with a broad caspase inhibitor. In the study, we also found that a higher dose of CsA induces LDH release when the cells were incubated for a longer period. Taken together, these data suggest that the mode of cell death induced by CsA is dose- and time-dependent. Short-term incubation with lower doses of CsA arrests cell growth; this arrest overlaps with the occurrence of apoptosis and then with necrosis after longer treatment periods with higher doses of CsA. (c) 2006 Elsevier Ltd. All rights reserved.