title: [Expression of cytosolic PrP and analysis of its cytotoxic activities].
abstract: 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.
authors.affiliation: College of Animal Husbandry and Veterinary Medicine, Shenyang Agricultural University, Shenyang 110161, China. wangxin82-82@163.com
authors.forename: Xin
authors.initials: X
authors.lastname: Wang
authors.forename: Chen-fang
authors.initials: CF
authors.lastname: Dong
authors.forename: Qi
authors.initials: Q
authors.lastname: Shi
authors.forename: Song
authors.initials: S
authors.lastname: Shi
authors.forename: Gui-rong
authors.initials: GR
authors.lastname: Wang
authors.forename: Yan-jun
authors.initials: YJ
authors.lastname: Lei
authors.forename: Run
authors.initials: R
authors.lastname: An
authors.forename: Kun
authors.initials: K
authors.lastname: Xu
authors.forename: Hui-ying
authors.initials: HY
authors.lastname: Jiang
authors.forename: Jun
authors.initials: J
authors.lastname: Han
authors.forename: Yun-jun
authors.initials: YJ
authors.lastname: Zhao
authors.forename: Xiao-ping
authors.initials: XP
authors.lastname: Dong
mesh_terms: 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
chemical_list: D011328:Prions; D019286:Endopeptidase K