Hyperbaric oxygen enhances the expression of prion protein and heat shock protein 70 in a mouse neuroblastoma cell line.
1. Cellular prion protein, PrP(C), is a ubiquitous glycoprotein strongly expressed in neurons with an as yet unknown biological function. In previous studies, we demonstrated that PrP(C) could be regulated by heat shock stress, implying that it might be a stress-responsive protein. Hyperbaric oxygen (HBO) administration is a well-defined model for the study of oxidative stress. 2. This study investigated the effect of HBO on PrP(C) and Hsp 70 expression in mouse neuroblastoma cell lines (N18), assessing the expression of PrP(C) and Hsp 70 using RT-PCR and Western blotting. HBO administration resulted in a time- and dose-dependent increase in PrP(C) and Hsp70 expression in N18 cells at both mRNA and protein levels, with a concomitant upregulation of c-Jun N-terminal kinase (JNK). 3. Under HBO treatment, luciferase reporter constructs of the rat PrP(C) promoter, containing the heat shock element (HSE) also present in Hsp70, expressed higher luciferase activity (3- to 10-fold) than those constructs without HSE. 4. In summary, these data suggest that PrP(C) and Hsp 70 may be regulated by HBO, through the activation of JNK. Thus, the activated heat shock transcriptional factor 1, phosphorylated by JNK interacted with HSE in the promoter of PrP(C) resulted in increased gene expression. These findings are vital for future therapeutic approaches in transmissible spongiform encephalopathies and the understanding of the function of the PrP(C).
Shyu
Woei-Cherng
WC
Neuro-Medical Scientific Center, Tzu-Chi Buddhist General Hospital, Tzu-Chi University, Hualien, Taiwan, Republic of China. shyu9423@sinamail.com
Lin
Shinn-Zong
SZ
Saeki
Keiichi
K
Kubosaki
Astsutaka
A
Matsumoto
Yoshitsugu
Y
Onodera
Takashi
T
Chiang
Ming-Fu
MF
Thajeb
Peterus
P
Li
Hung
H
D000818:Animals; D001921:Brain / Q000378:metabolism / Q000503:physiopathology; D045744:Cell Line, Tumor; D004268:DNA-Binding Proteins / Q000187:drug effects / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics; D018840:HSP70 Heat-Shock Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D000076249:Heat Shock Transcription Factors; D006931:Hyperbaric Oxygenation; D048031:JNK Mitogen-Activated Protein Kinases; D008156:Luciferases / Q000235:genetics; D051379:Mice; D020928:Mitogen-Activated Protein Kinases / Q000187:drug effects / Q000378:metabolism; D009447:Neuroblastoma; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D018384:Oxidative Stress / Q000187:drug effects / Q000502:physiology*; D010100:Oxygen / Q000494:pharmacology*; D010766:Phosphorylation; D018621:PrPC Proteins / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000187:drug effects / Q000235:genetics; D012333:RNA, Messenger / Q000187:drug effects / Q000378:metabolism; D013997:Time Factors; D014157:Transcription Factors; D015854:Up-Regulation / Q000187:drug effects / Q000502:physiology
D004268:DNA-Binding Proteins; D018840:HSP70 Heat-Shock Proteins; D000076249:Heat Shock Transcription Factors; D018621:PrPC Proteins; D012333:RNA, Messenger; D014157:Transcription Factors; D008156:Luciferases; D048031:JNK Mitogen-Activated Protein Kinases; D020928:Mitogen-Activated Protein Kinases; D010100:Oxygen