Klacanova Katarina, Pilchova Ivana, Klikova Katarina, Racay Peter
Department of Medical Biochemistry and Division Neuroscience of Biomedical Center, Jessenius Faculty of Medicine in Martin, Comenius University in Bratislava, Mala Hora 4D, Martin, SK-03601, Slovak Republic.
J Mol Neurosci. 2016 Apr;58(4):497-506. doi: 10.1007/s12031-015-0685-4. Epub 2015 Nov 19.
Both translation arrest and proteasome stress associated with accumulation of ubiquitin-conjugated protein aggregates were considered as a cause of delayed neuronal death after transient global brain ischemia; however, exact mechanisms as well as possible relationships are not fully understood. The aim of this study was to compare the effect of chemical ischemia and proteasome stress on cellular stress responses and viability of neuroblastoma SH-SY5Y and glioblastoma T98G cells. Chemical ischemia was induced by transient treatment of the cells with sodium azide in combination with 2-deoxyglucose. Proteasome stress was induced by treatment of the cells with bortezomib. Treatment of SH-SY5Y cells with sodium azide/2-deoxyglucose for 15 min was associated with cell death observed 24 h after treatment, while glioblastoma T98G cells were resistant to the same treatment. Treatment of both SH-SY5Y and T98G cells with bortezomib was associated with cell death, accumulation of ubiquitin-conjugated proteins, and increased expression of Hsp70. These typical cellular responses to proteasome stress, observed also after transient global brain ischemia, were not observed after chemical ischemia. Finally, chemical ischemia, but not proteasome stress, was in SH-SY5Y cells associated with increased phosphorylation of eIF2α, another typical cellular response triggered after transient global brain ischemia. Our results showed that short chemical ischemia of SH-SY5Y cells is not sufficient to induce both proteasome stress associated with accumulation of ubiquitin-conjugated proteins and stress response at the level of heat shock proteins despite induction of cell death and eIF2α phosphorylation.
翻译阻滞和与泛素结合蛋白聚集体积累相关的蛋白酶体应激均被视为短暂性全脑缺血后神经元延迟死亡的原因;然而,确切机制以及可能的关系尚未完全明确。本研究的目的是比较化学性缺血和蛋白酶体应激对神经母细胞瘤SH-SY5Y细胞和胶质母细胞瘤T98G细胞的细胞应激反应及活力的影响。化学性缺血通过用叠氮化钠联合2-脱氧葡萄糖短暂处理细胞来诱导。蛋白酶体应激通过用硼替佐米处理细胞来诱导。用叠氮化钠/2-脱氧葡萄糖处理SH-SY5Y细胞15分钟与处理后24小时观察到的细胞死亡相关,而胶质母细胞瘤T98G细胞对相同处理具有抗性。用硼替佐米处理SH-SY5Y细胞和T98G细胞均与细胞死亡、泛素结合蛋白的积累以及Hsp70表达增加相关。这些在短暂性全脑缺血后也观察到的对蛋白酶体应激的典型细胞反应,在化学性缺血后未观察到。最后,化学性缺血而非蛋白酶体应激在SH-SY5Y细胞中与eIF2α磷酸化增加相关,eIF2α磷酸化是短暂性全脑缺血后触发的另一种典型细胞反应。我们的结果表明,SH-SY5Y细胞的短期化学性缺血不足以诱导与泛素结合蛋白积累相关的蛋白酶体应激以及热休克蛋白水平的应激反应,尽管诱导了细胞死亡和eIF2α磷酸化。