转染hSOD1-G93A对NSC-34细胞内铁代谢相关蛋白表达的影响  

Effect of transfection of hSOD1-G93A on expressions of iron-related proteins in NSC-34 cells

作　　者：王寿刚[1] 陆玉成[2] 黄丽梅[3] 尤翠平[2] 王龙[2] 王富敏[5] 苏全平[1] 于继徐[4] 车峰远[4] 

机构地区：[1]泰山医学院,泰安271016 [2]临沂市人民医院中心实验室,276003 [3]临沂市人民医院急诊科,276003 [4]临沂市人民医院神经内科,276003 [5]山东省立医院睡眠医学中心,济南250021

出　　处：《中华神经医学杂志》2015年第9期905-909,共5页Chinese Journal of Neuromedicine

基　　金：山东省自然科学基金（ZR2010HM041）;山东省博士后创新项目专项资金（201102004）

摘　　要：目的探讨转染hSODl．G93A对NSC-34细胞内铁代谢相关蛋白表达的影响。方法常规体外培养未转染、转染pcDNA3．1（-）、hSODl-pcDNA3．1（-）和hSODl-G93A—pcDNA3．1（-）质粒的NSC-34细胞系；分为正常组、空转组、野生组和突变组。利用免疫组化染色法观察各组细胞的形态变化；通过检测丙二醛（MDA）含量观察各组细胞氧化应激水平；用逆转录聚合酶链式反应rRT．PCR）和Westernblotting分别检测各组细胞内铁代谢相关蛋白的基因和蛋白表达水平。结果突变组的细胞胞体变圆，突起减少、变短，细胞内MDA含量明显高于其他3组细胞，差异有统计学意义（P〈0．05）；各组细胞内的转铁蛋白受体（TfR）、二价金属离子转运体1（DMTI）、铁蛋白等基因在基因水平上表达差异无统计学意3L（P〈0．05）；各组细胞内的TfR、DMTl、铁蛋白等在蛋白水平上表达差异无统计学意义（胗O．05），但是突变组细胞铁蛋白表达较正常组细胞明显升高，差异有统计学意义（P，0．051。结论转染hSOD1-G93A对NSC-34细胞（突变组）内TfR、DMTl表达无明显影响．但可以使细胞内铁蛋白在蛋白表达水平明显升高。Objective To investigate the effect of transfection of hSOD 1-G93A on expression of iron-related proteins in NSC-34 cells. Methods The NSC-34 cell lines from four groups （without transfection or stably transfected with pcDNA3.1 （-）, hSOD 1 -pcDNA3.1 （-） and hSOD 1 -G93A-pcDNA3.1 （-） plasmids） were cultured at routine ways. Morphological changes of the transfected cells were observed by immuno- histochemical method. The cell oxidative damage degrees were observed by detecting malondialdehyde content in cells. Real time-PCR was used to determine the mRNA expression levels of iron- related genes; Western blotting was used to determine the changes of protein levels of iron- related genes. Results The soma became round and the number of neurite decreased in the NSC-34 cells transfected with hSOD1-G93A-pcDNA3, i （-） plasmid, the neurites were shorter and oxidative stress was increased as compared with those in the other three cell lines （P〈0.05）. The mRNA expressions of transferrin receptor （TfR）, divalent metal ion 1 （DMT1）, and ferritin were not significantly different among the four groups （P〉0.05）; the protein expressions of TfR and DMT1 were not significantly different among the four groups （P〉0.05）, while that of ferritin in the NSC-34 cells transfected with hSOD1-G93A-pcDNA3.1 （-） plasmid was significantly higher than that in cells without transfection （P〉 0.05）. Conclusion The expressions of TfR and DMT1 are not influenced by hSOD1-G93A but the expression of ferritin in the mutant cells are increased by the influence ofhSOD1-G93A.

关 键 词：肌萎缩侧索硬化 NSC-34细胞 转铁蛋白受体 二价金属离子转运体1 

分 类 号：R744.8[医药卫生—神经病学与精神病学]