Overexpression of human peroxiredoxin 5 in subcellular compartments of Chinese hamster ovary cells: Effects on cytotoxicity and DNA damage caused by peroxides
Overexpression of human peroxiredoxin 5 in subcellular compartments of Chinese hamster ovary cells: Effects on cytotoxicity and DNA damage caused by peroxides
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DOI：
10.1016/j.freeradbiomed.2003.10.019
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发表时间：
2004-01-01
影响因子：
7.4
通讯作者：
Knoops, B
中科院分区：
文献类型：
--
作者：
Banmeyer, I;Marchand, C;Knoops, B
Peroxiredoxin 5 是一种在组织中广泛表达的哺乳动物硫氧还蛋白过氧化物酶。过氧化还原蛋白 5 可在细胞内定位于线粒体、过氧化物酶体、细胞质，以及较小程度的细胞核。与其他五种哺乳动物过氧化还原蛋白相比，这种非常广泛的亚细胞分布促使我们根据其亚细胞定位进一步研究过氧化还原蛋白5在哺乳动物细胞中的抗氧化保护功能。通过稳定转染建立在细胞质、线粒体或细胞核中过表达人过氧化还原蛋白5的中国仓鼠卵巢细胞。将过氧化还原蛋白 5 过表达的细胞暴露于外源添加的过氧化氢或叔丁基氢过氧化物的低或急性氧化应激下 1 小时。通过克隆形成和乳酸脱氢酶测定评估过氧化还原蛋白 5 赋予的细胞保护作用。在细胞质、线粒体或核区室中过氧化还原蛋白 5 过表达可显着减少细胞死亡，在线粒体中过氧化还原蛋白 5 过表达可提供更有效的保护，证实该细胞器是过氧化物的主要靶标。此外，我们还通过彗星试验评估了过氧化氢或叔丁基过氧化氢诱导的核 DNA 损伤。细胞核中过氧化还原蛋白 5 的过度表达显着减少了两种过氧化物诱导的 DNA 损伤。总之，本研究表明，哺乳动物细胞中过氧化还原蛋白 5 的多个亚细胞靶向可能与非病理条件下的抗氧化保护机制有关，而且与病理生理情况下发生的过氧化物引起的急性氧化应激有关。 (C) 2003 Elsevier Inc. 保留所有权利。
Peroxiredoxin 5 is a mammalian thioredoxin peroxidase ubiquitously expressed in tissues. Peroxiredoxin 5 can be intracellularly localized to mitochondria, peroxisomes, the cytosol, and, to a lesser extent, the nucleus. This remarkably wide subcellular distribution compared with the five other mammalian peroxiredoxins prompted us to further investigate the antioxidant protective function of peroxiredoxin 5 in mammalian cells according to its subcellular localization. Chinese hamster ovary cells overexpressing human peroxiredoxin 5 in the cytosol, in mitochondria, or in the nucleus were established by stable transfection. Cells overexpressing peroxiredoxin 5 were exposed for 1 h to low or acute oxidative stress with exogenously added hydrogen peroxide or tert-butylhydroperoxide. Cell protection conferred by peroxiredoxin 5 was evaluated by clonogenicity and lactate dehydrogenase assays. Overexpressing peroxiredoxin 5 in either the cytosolic, mitochondrial, or nuclear compartment significantly reduced cell death, with more effective protection with overexpression of peroxiredoxin 5 in mitochondria, confirming that this organelle is a major target of peroxides. Moreover, we evaluated, with the comet assay, nuclear DNA damage induced by hydrogen peroxide or tert-butylhydroperoxide. Overexpression of peroxiredoxin 5 in the nucleus significantly decreased DNA damage induced by both peroxides. In conclusion, the present study suggests that multiple subcellular targeting of peroxiredoxin 5 in mammalian cells can be implicated in antioxidant protective mechanisms under nonpathological conditions but also during acute oxidative stress caused by peroxides occurring in pathophysio logical situations. (C) 2003 Elsevier Inc. All rights reserved.