An Run, Chu Yong-lie, Tian Chan, Dai Xiao-xia, Chen Jing-hong, Shi Qi, Han Jun, Dong Xiao-ping
School of Medicine, Xi'an Jiao-Tong University, Xi'an 710061, China.
Acta Pharmacol Sin. 2008 Dec;29(12):1451-8. doi: 10.1111/j.1745-7254.2008.00902.x.
To determine whether the antitumor factor nm23 is related with antioxidation.
Full-length human nm23-H1 was cloned into a mammalianexpressing vector and transiently introduced into HeLa cells.
A remarkably low level of reactive oxygen species (ROS) was detected in the cells overexpressing nm23-H1. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and trypan blue assays found that the cells transfected with a nm23- H1-expressing plasmid had higher viability and stronger resistance to oxidative stress. Immunoprecipitation tests revealed that endogenous nm23-H1 formed a protein complex with p53. Furthermore, the intracellular levels of p53 and p53- regulated gene GPX1 were obviously increased in the cells overexpressing nm23- H1. The downregulation of p53 in the cells overexpressing nm23-H1 resulted in a higher cellular ROS level and lower cell viability.
The findings suggest that nm23-H1 may act as a cellular protector against oxidative stress, possibly triggering the p53-related antioxidative pathway.
确定抗肿瘤因子nm23是否与抗氧化作用相关。
将人nm23-H1全长克隆至哺乳动物表达载体中,并瞬时导入HeLa细胞。
在过表达nm23-H1的细胞中检测到极低水平的活性氧(ROS)。噻唑蓝和台盼蓝检测发现,转染了表达nm23-H1质粒的细胞具有更高的活力和更强的抗氧化应激能力。免疫沉淀试验显示,内源性nm23-H1与p53形成了蛋白复合物。此外,在过表达nm23-H1的细胞中,p53及p53调控基因GPX1的细胞内水平明显升高。在过表达nm23-H1的细胞中下调p53会导致细胞内ROS水平升高和细胞活力降低。
这些发现表明,nm23-H1可能作为细胞对抗氧化应激的保护因子,可能触发与p53相关的抗氧化途径。