Overexpression of glia maturation factor in C6 cells promotes differentiation and activates superoxide dismutase
Overexpression of glia maturation factor in C6 cells promotes differentiation and activates superoxide dismutase
复制标题
DOI：
10.1023/a:1020715126326
复制
发表时间：
1998-11-01
影响因子：
4.4
通讯作者：
Oberley, LW
中科院分区：
文献类型：
--
作者：
Lim, R;Zaheer, A;Oberley, LW
为了评价胶质细胞成熟因子（GMF）的细胞内功能，我们过表达GMF在C6大鼠胶质瘤细胞中使用两种方法：稳定转染使用pcDNA3质粒，和瞬时转染使用复制缺陷型人腺病毒。使用这两种方法，与未转染或仅载体对照相比，用GMF转染并过表达蛋白质的C6细胞在培养物中表现出较低的饱和密度。转染的细胞还表现出形态分化，如细胞突起的生长所示。当接种到裸鼠体内时，转染细胞的致瘤性低于对照组，并表达成熟的星形胶质细胞标志物胶质细胞酸性蛋白。在组织培养中，转染的细胞显示CuZn依赖性超氧化物歧化酶（CuZnSOD）活性增加3.5倍。Western印迹分析显示CuZnSOD蛋白增加了3.5倍，表明该酶的诱导。鉴于最近的研究发现，活性氧（ROS）和抗氧化酶是错综复杂地参与关键的生理过程，如增殖，分化和凋亡，该研究提出了可能性，CuZnSOD可能是一个介导的转基因食品的功能。
In order to evaluate the intracellular function of glia maturation factor (GMF), we overexpressed GMF in C6 rat glioma cells using two methods: stable transfection using the pcDNA3 plasmid, and transient transfection using replication-defective human adenovirus. With both methods, C6 cells transfected with GMF and overexpressing the protein exhibit a lower saturation density in culture compared to non-transfected or vector alone controls. Transfected cells also exhibit morphological differentiation as shown by the outgrowth of cell processes. When inoculated into nude mice, transfected cells are less tumorigenic than controls, and express the mature astrocytic marker glial fibrillary acidic protein. In tissue culture, transfected cells show a 3.5-fold increase in CuZn-dependent superoxide dismutase (CuZnSOD) activity. Western blot analysis reveals a 3.5-fold increase in CuZnSOD protein, suggesting an induction of the enzyme. In view of recent findings that reactive oxygen species (ROS) and the antioxidant enzymes are intricately involved in key physiologic processes such as proliferation, differentiation and apoptosis, the study raises the possibility that CuZnSOD may be a mediator of GMF function.