[Expression of cytosolic PrP and analysis of its cytotoxic activities].
In order to study the physicochemical characteristics of cytosolic PrP (CytoPrP) and evaluate its possible influence on cell viability, a recombinant plasmid expressing human CytoPrP eukaryoticly was constructed and transfected into human neuroblastoma cell line SH-SY5Y transiently. Proteinase-resistant activities of CytoPrP were evaluated by a proteinase K (PK) digestion and cytotoxic effects of CytoPrP were tested by MTT assay and Trypan Blue cell-counting. The presence of CytoPrP in cytoplasm after transfection was controlled by the presence of protease inhibitor. Compared with wild-type PrP, CytoPrP possessed relatively stronger PK-resistant activities. Obvious cytotoxic effects were observed in the cells after inducement of CytoPrP in cytoplasm by protease inhibitor, showing a dose-dependent manner. The results provide useful scientific evidences for further studies of potential role of CytoPrP in pathological mechanism of prion disease.
Wang
Xin
X
College of Animal Husbandry and Veterinary Medicine, Shenyang Agricultural University, Shenyang 110161, China. wangxin82-82@163.com
Dong
Chen-fang
CF
Shi
Qi
Q
Shi
Song
S
Wang
Gui-rong
GR
Lei
Yan-jun
YJ
An
Run
R
Xu
Kun
K
Jiang
Hui-ying
HY
Han
Jun
J
Zhao
Yun-jun
YJ
Dong
Xiao-ping
XP
D045744:Cell Line, Tumor; D002470:Cell Survival; D003600:Cytosol / Q000737:chemistry; D019286:Endopeptidase K / Q000494:pharmacology; D006801:Humans; D011328:Prions / Q000235:genetics / Q000502:physiology*; D014162:Transfection
D011328:Prions; D019286:Endopeptidase K