Department of Neurology, College of Medicine, Hanyang University, #17 Haengdang-dong, Seongdong-gu, Seoul 139-791, Korea.
Mol Cell Biochem. 2010 Jul;340(1-2):133-41. doi: 10.1007/s11010-010-0410-0. Epub 2010 Feb 20.
Human angiogenin (ANG) has been highlighted as an angiogenic factor which supports primary and metastatic tumor growth. Recent genetic studies have shown that ANG is presented as a susceptibility gene for amyotrophic lateral sclerosis (ALS) and ALS-frontotemporal dementia (ALS-FTD). They found several missense mutations, including K40I, which present the weakest functional activity in ANG variants. In this study, we investigate whether human wild type ANG (wANG) and its variant K40I (mANG) maintain their divergent functional capacities in neuronal cells. To evaluate this, SH-SY5Y neuroblastoma cells were transfected with wANG and mANG DNA and identified both wild and mutant ANG are localized to nuclei and have no effects on proliferation. We have shown that human wANG prevented cell death under H(2)O(2)-induced oxidative stress in both SH-SY5Y and NSC-34 cells, tested by MTT assay. These effects were more enhanced in motor neuron cell NSC-34. wANG also played a role in cell migration, while mANG decreased these functional activities. Immunoblot analysis revealed that the intracellular signaling of ERK1/2 (at Thr183/Tyr185) was increased following transfection of the wANG gene, and significantly decreased by mANG in neuronal cells. These findings suggest that human ANG plays a critical role in cell protection and migration following alterations in ERK1/2 signaling in SH-SY5Y cells. This may provide the possible relationship between mutations in hANG and other neurodegenerative diseases as well as ALS.
人血管生成素（ANG）已被强调为支持原发性和转移性肿瘤生长的血管生成因子。最近的遗传研究表明，ANG 是肌萎缩侧索硬化症（ALS）和 ALS-额颞叶痴呆（ALS-FTD）的易感基因。他们发现了几个错义突变，包括 K40I，其在 ANG 变体中表现出最弱的功能活性。在这项研究中，我们研究了人野生型 ANG（wANG）及其变体 K40I（mANG）是否在神经元细胞中保持其不同的功能能力。为了评估这一点，用 wANG 和 mANG DNA 转染 SH-SY5Y 神经母细胞瘤细胞，并确定野生和突变 ANG 都定位于细胞核，对增殖没有影响。我们已经表明，人 wANG 通过 MTT 测定在 SH-SY5Y 和 NSC-34 细胞中预防 H2O2 诱导的氧化应激引起的细胞死亡。在运动神经元细胞 NSC-34 中，这些效果得到了增强。wANG 还在细胞迁移中起作用，而 mANG 降低了这些功能活性。免疫印迹分析显示，wANG 基因转染后 ERK1/2（在 Thr183/Tyr185）的细胞内信号增加，而在神经元细胞中 mANG 则显著降低。这些发现表明，人 ANG 在 SH-SY5Y 细胞中 ERK1/2 信号改变后，在细胞保护和迁移中发挥关键作用。这可能为 hANG 突变与其他神经退行性疾病以及 ALS 之间的可能关系提供依据。
