人神经母细胞瘤SH-SY5Y细胞差异分泌氧化应激报告生物标志物的蛋白质组学鉴定

T. Toda, Megumi Nakamura, H. Morisawa, M. Hirota
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引用次数: 2

摘要

自由基理论预测,氧化应激加速了衰老的速度,增加了老年人退行性疾病的发病。多巴胺能神经元由于在多巴胺代谢途径中产生活性氧而特别容易受到与年龄相关的神经元疾病的影响。帕金森病患者黑质发生的生化变化提示氧化应激诱导的细胞损伤可能参与了神经退行性变。在我们之前的研究中,我们发现伸长因子-2的去磷酸化和核纤层蛋白A/C的磷酸化可能是神经元细胞对氧化应激的特异性反应。(Nakamura等人)BBA, 1763(9), 977-989, 2006)这些蛋白的去磷酸化和磷酸化是分析应激反应分子机制的重要生物标志物,然而,如果在患者的脑脊液或血清中检测不到,这种磷酸化蛋白质组分析被认为不适合用于神经变性的临床研究。因此,我们使用对人SH-SY5Y神经母细胞瘤细胞施加氧化应激的培养系统,对分泌蛋白的蛋白质组进行2D-DIGE分析。根据分泌组分析结果,我们确定了泛素激活酶E1、泛素结合酶E2 N、泛素c端水解酶l1、14-3-3蛋白异构体、Rab GDP解离抑制剂β、Rho GDP解离抑制剂1、过氧化物还毒素2、谷胱甘肽s转移酶P、α烯醇化酶、LDH B链作为氧化应激报告的生物标志物。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Proteomic identification of oxidative-stress-reporting biomarkers differentially secreted from human neuroblastoma SH-SY5Y cells
The free-radical theory predicts that the oxidative stress accelerates the rate of aging and increases the onset of degenerative disorders in the elderly. Dopaminergic neurons are especially vulnerable to age-related neuronal disorders due to reactive oxygen species generated in the pathway of dopamine metabolism. Biochemical changes occurring in substantia nigra of Parkinson's disease patients suggest that the oxidative-stress-induced cell damages may be involved in the neurodegeneration. In our previous researches, we found that the dephosphorylation of elongation factor-2 and phosphorylation of nuclear lamin A/C might be neuronal cell specific response to oxidative stress. (Nakamura et al. BBA, 1763(9), 977-989, 2006) The dephosphorylation and phosphorylation of those proteins are significant biomarkers for analyzing the molecular mechanisms of the stress response, however, such a phosphoproteome analysis is thought to be inappropriate for clinical investigation of neurodegeneration if it was not detectable in cerebrospinal fluid or serum of patients. Thus, we proceeded to the 2D-DIGE analysis of secretome, proteome of secreted proteins, using the culture system in which oxidative stress was applied to human SH-SY5Y neuroblastoma cells. As the result of our secretome analysis, we identified ubiquitin-activating enzyme E1, ubiquitin-conjugating enzyme E2 N, ubiquitin C-terminal hydrorase-L1, 14-3-3 protein isoforms, Rab GDP dissociation inhibitor β, Rho GDP-dissociation inhibitor 1, peroxiredoxin-2, glutathione S-transferase P, α enolase, LDH B chain as oxidative-stress-reporting biomarker candidates.
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