G_1-Dependent Prion Protein Expression in Human Glioblastoma Cell Line T98G

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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 G_1 phase.
社団法人日本薬学会の論文
2002-06-01