G1-Dependent Prion Protein Expression in Human Glioblastoma Cell Line T98G

• Published in: Biological & pharmaceutical bulletin Vol. 25; no. 6; pp. 728 - 733
• Main Authors: Kikuchi, Yutaka, Kakeya, Tomoshi, Yamazaki, Takeshi, Takekida, Kaori, Nakamura, Naoto, Matsuda, Haruo, Takatori, Kosuke, Tanimura, Akio, Tanamoto, Ken-ichi, Sawada, Jun-ichi
• Format: Journal Article
• Language: English
• Published: Tokyo The Pharmaceutical Society of Japan 01.06.2002; Maruzen; Japan Science and Technology Agency
• Subjects: Biological and medical sciences; Brain Neoplasms - metabolism; cell cycle; cell density; cellular prion protein; Culture Media, Conditioned; Culture Media, Serum-Free; Degenerative and inherited degenerative diseases of the nervous system. Leukodystrophies. Prion diseases; DNA - biosynthesis; DNA Primers; Glioblastoma - metabolism; Humans; Immunoblotting; Medical sciences; Neurology; Neuropharmacology; Neuroprotective agent; Nuclear Proteins - metabolism; Pharmacology. Drug treatments; Prion Proteins; Prions - metabolism; PrPC Proteins - biosynthesis; Reverse Transcriptase Polymerase Chain Reaction; RNA - biosynthesis; RNA - isolation & purification; T98G cell; Time Factors; Tumor Cells, Cultured; Human; Glial cell; G1 Phase; Prion disease; Neuroglia; Established cell line; Neuroprotective agent; Mechanism of action; Prion protein; In vitro; Biological activity; Tumor cell
• ISSN: 0918-6158; 1347-5215
• DOI: 10.1248/bpb.25.728

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.