Song Zhi-Qi, Yang Li-Feng, Wang Yun-Sheng, Zhu Ting, Zhou Xiang-Mei, Yin Xiao-Min, Yao Hong-Qiang, Zhao De-Ming
State Key Laboratories for Agrobiotechnology, Key Lab of Animal Epidemiology and Zoonosis, Ministry of Agriculture, National Animal Transmissible Spongiform Encephalopathy Laboratory, College of Veterinary Medicine, China Agricultural University, Beijing, China.
CNS Neurosci Ther. 2014 Aug;20(8):737-47. doi: 10.1111/cns.12243. Epub 2014 Mar 15.
Prion diseases are a group of infectious neurodegenerative diseases characterized by neuronal death and degeneration. Human leukocyte antigen-B-associated transcript 3 (BAT3) is an important apoptosis regulator. We therefore investigated the interactions between BAT3 and prion protein and the potential role of BAT3 in PrP106-126-induced apoptosis.
BAT3 and prion protein were overexpressed in Hela, Neuro2A, or primary neuronal cells by transfection with BAT3-HA or PRNP-EGFP expression plasmids and their relationship studied by immunofluorescence and Western blotting. The effect of BAT3 on PrP106-126-induced cytotoxicity and apoptosis was detected by the CCK-8 assay and terminal-deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) assay. The expression of cytochrome c and Bcl-2 was examined by Western blotting.
BAT3 interacted with prion protein and enhanced PrP expression. After PrP106-126 peptide treated, BAT3 was transported from the nucleus to cytoplasm, increased cell viability, and protected neurons from PrP106-126-induced apoptosis through stabilizing the level of Bcl-2 protein and inhibiting the release of cytochrome c to cytoplasm.
Our present data showed a novel molecular mechanism of PrP106-126-induced apoptotic process regulation through the overexpression of BAT3, which may be important for the basic regulatory mechanism of neuron survival in prion diseases and associated neurodegenerative diseases in vivo.
朊病毒病是一组以神经元死亡和变性为特征的传染性神经退行性疾病。人类白细胞抗原B相关转录本3（BAT3）是一种重要的凋亡调节因子。因此，我们研究了BAT3与朊病毒蛋白之间的相互作用以及BAT3在PrP106 - 126诱导的凋亡中的潜在作用。
通过用BAT3 - HA或PRNP - EGFP表达质粒转染，在Hela细胞、Neuro2A细胞或原代神经元细胞中过表达BAT3和朊病毒蛋白，并通过免疫荧光和蛋白质印迹法研究它们之间的关系。通过CCK - 8法和末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）法检测BAT3对PrP106 - 126诱导的细胞毒性和凋亡的影响。通过蛋白质印迹法检测细胞色素c和Bcl - 2的表达。
BAT3与朊病毒蛋白相互作用并增强PrP表达。在PrP106 - 126肽处理后，BAT3从细胞核转运至细胞质，提高细胞活力，并通过稳定Bcl - 2蛋白水平和抑制细胞色素c释放到细胞质中来保护神经元免受PrP106 - 126诱导的凋亡。
我们目前的数据显示了通过BAT3过表达调节PrP106 - 126诱导的凋亡过程的新分子机制，这可能对朊病毒病及体内相关神经退行性疾病中神经元存活的基本调节机制具有重要意义。
