Document Detail

G1-dependent prion protein expression in human glioblastoma cell line T98G.
MedLine Citation:
PMID:  12081137     Owner:  NLM     Status:  MEDLINE    
Human glioblastoma cell line T98G produced a cellular form of prion protein (PrP(C)), and we confirmed expression of PrP mRNA by RT-PCR. Immunoblot analysis of whole cell lysate revealed one major (35 kDa) and two faint bands (31, 25 kDa) that reacted with monoclonal anti-human PrP antibody 3F4. Cells treated with tunicamycin produced only a 25 kDa band, representing a deglycosylated form of PrP. Similarly, peptide: N-glycosidase F treatment of whole cell lysate altered the Asn-linked form to the deglycosylated form. When T98G cells were cultured for a longer period, the amount of PrP(C) per cell increased on Day 4 to 16 in a time-dependent manner. When the cells were cultured at high cell-density, the cells on Day 4 produced the same amount of PrP(C) as those on Day 16 of the usual culture. Moreover, in a serum-free medium, cells cultured at a low cell-density produced the same amount of PrP(C) as those cultured at the high cell-density. These results demonstrate that PrP(C) production in T98G cells was dependent on the phase of the cell cycle, probably the G1 phase.
Yutaka Kikuchi; Tomoshi Kakeya; Takeshi Yamazaki; Kaori Takekida; Naoto Nakamura; Haruo Matsuda; Kosuke Takatori; Akio Tanimura; Ken-ichi Tanamoto; Jun-ichi Sawada
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biological & pharmaceutical bulletin     Volume:  25     ISSN:  0918-6158     ISO Abbreviation:  Biol. Pharm. Bull.     Publication Date:  2002 Jun 
Date Detail:
Created Date:  2002-06-25     Completed Date:  2002-12-31     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  9311984     Medline TA:  Biol Pharm Bull     Country:  Japan    
Other Details:
Languages:  eng     Pagination:  728-33     Citation Subset:  IM    
Division of Microbiology, National Institute of Health Sciences, Tokyo, Japan.
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MeSH Terms
Brain Neoplasms / metabolism*
Culture Media, Conditioned
Culture Media, Serum-Free
DNA / biosynthesis
DNA Primers
Glioblastoma / metabolism*
Nuclear Proteins / metabolism*
PrPC Proteins / biosynthesis*
Prions / metabolism*
RNA / biosynthesis,  isolation & purification
Reverse Transcriptase Polymerase Chain Reaction
Time Factors
Tumor Cells, Cultured
Reg. No./Substance:
0/Culture Media, Conditioned; 0/Culture Media, Serum-Free; 0/DNA Primers; 0/Nuclear Proteins; 0/PRNP protein, human; 0/PrPC Proteins; 0/Prions; 63231-63-0/RNA; 9007-49-2/DNA

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