NR AGIQ
AU Kikuchi,Y.; Kakeya,T.; Yamazaki,T.; Takekida,K.; Nakamura,N.; Matsuda,H.; Takatori,K.; Tanimura,A.; Tanamoto,K.; Sawada,J.
TI G1-dependent prion protein expression in human glioblastoma cell line T98G
QU Biological and Pharmaceutical Bulletin 2002 Jun; 25(6): 728-33
PT journal article
AB Human glioblastoma cell line T98G produced a cellular form of prion protein (PrPc), 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 PrPc 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 PrPc 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 PrPc as those cultured at the high cell-density. These results demonstrate that PrPc production in T98G cells was dependent on the phase of the cell cycle, probably the G1 phase.
AD Division of Microbiology, National Institute of Health Sciences, Tokyo, Japan. kikuchi@nihs.go.jp
SP englisch
PO Japan