Overexpression of mutated Cu,Zn-SOD in neuroblastoma cells results in cytoskeletal change
Overexpression of mutated Cu,Zn-SOD in neuroblastoma cells results in cytoskeletal change
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DOI：
10.1152/ajpcell.00014.2004
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发表时间：
2005-02-01
影响因子：
5.5
通讯作者：
Taniguchi, N
中科院分区：
文献类型：
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作者：
Takamiya, R;Takahashi, M;Taniguchi, N
肌萎缩侧索硬化症(ALS)涉及脊髓和运动皮质运动神经元的进行性变性。家族性肌萎缩侧索硬化症(FAL)患者中有15-20%的患者存在编码铜锌超氧化物歧化酶(SOD)的SOD1基因缺陷。为了阐明突变的铜锌超氧化物歧化酶的病理作用，我们研究了突变的铜锌超氧化物歧化酶是否影响细胞周期的问题。将人野生型和突变型(G37R，G93A)铜锌超氧化物歧化酶分别导入小鼠神经母细胞瘤Neuro-2a细胞。铜锌超氧化物歧化酶基因突变后，细胞生长明显减慢，细胞周期发生G(2)/M期阻滞。它们对鬼臼毒素的反应性也较低，这表明细胞骨架被破坏。免疫沉淀、双向凝胶电泳法和Western印迹分析表明，突变的铜锌超氧化物歧化酶与肌动蛋白结合。纯化的肌动蛋白和突变的铜锌超氧化物歧化酶(G93A)的体外孵育实验也得到了类似的结果。这些结果表明，FALS中突变的铜锌超氧化物歧化酶通过与肌动蛋白结合引起细胞骨架改变，从而导致G(2)/M期停滞和生长迟缓。
Amyotrophic lateral sclerosis (ALS) involves the progressive degeneration of motor neurons in the spinal cord and the motor cortex. It has been shown that 15-20% of patients with familial ALS (FALS) have defects in the Sod1 gene, which encodes Cu, Zn-superoxide dismutase ( SOD). To elucidate the pathological role of mutated Cu, Zn-SOD, we examined the issue of whether mutated Cu, Zn-SOD affects the cell cycle. Mouse neuroblastoma Neuro-2a cells were transfected with human wild-type or mutated (G37R, G93A) Cu, Zn-SOD. Mutated, Cu, Zn-SOD-transfected cells exhibited marked retardation in cell growth and G(2)/M arrest. They also displayed lower reactivity to phalloidin, indicating that the cytoskeleton was disrupted. Immunoprecipitation, two-dimensional gel electrophoresis, and Western blot analysis indicated that mutated Cu,Zn-SOD associates with actin. Similar results were obtained by in vitro incubation experiments with purified actin and mutated Cu,Zn-SOD (G93A). These results suggest that mutated Cu,Zn-SOD in FALS causes cytoskeletal changes by associating with actin, which subsequently causes G(2)/M arrest and growth retardation.