Glycosylation modification of human prion protein provokes apoptosis in HeLa cells in vitro.
We investigate the correlation between the glycosylation modified prion proteins and apoptosis. The wild-type PRNP gene and four PRNP gene glycosylated mutants were transiently expressed in HeLa cells. The effect of apoptosis induced by PrP mutants was confirmed by MTT assay, Hochest staining, Annexin-V staining and PI staining. ROS test detected ROS generation within the cells. The mitochondrial membrane potential was analyzed by the flow cytometry. The expression levels of Bcl-xL, Bax, cleaved Caspase-9 proteins were analyzed by Western Blot. The results indicated that the expressed non-glycosylated PrP in HeLa cells obviously induced apoptosis, inhibited the growth of cells and reduced the mitochondrial membrane potential, and more ROS generation and low levels of the apoptosis-related proteins Bcl-xL, the activated the cleaved Caspase-9 proteins were found. The apoptosis induced by non-glycosylated PrP demonstrates that its underlying mechanism correlates with the mitochondria-mediated signal transduction pathway.
Yang
Yang
Y
National Laboratory of Medical Molecular Biology, Department of Biochemistry and Molecular Biology, Institute of Basic Medical Science, Chinese Academy of Medical Sciences, Beijing 100005, People's Republic of China. caiminxu@yahoo.com.cn
Chen
Lan
L
Pan
Hua-Zhen
HZ
Kou
Yi
Y
Xu
Cai-Min
CM
D017209:Apoptosis / Q000502:physiology; D053453:Caspase 9 / Q000378:metabolism; D049109:Cell Proliferation; D002470:Cell Survival; D006031:Glycosylation; D006367:HeLa Cells; D006801:Humans; D050505:Mutant Proteins / Q000378:metabolism / Q000502:physiology; D011328:Prions / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*; D011499:Protein Processing, Post-Translational / Q000502:physiology; D017382:Reactive Oxygen Species / Q000378:metabolism; D014018:Tissue Distribution; D014162:Transfection; D051020:bcl-X Protein / Q000378:metabolism
C494811:BCL2L1 protein, human; D050505:Mutant Proteins; D011328:Prions; D017382:Reactive Oxygen Species; D051020:bcl-X Protein; D053453:Caspase 9