title: Characterization of a new human glioblastoma cell line that expresses mutant p53 and lacks activation of the PDGF pathway.
abstract: We have established and characterized a new glioblastoma cell line, termed GT9, from a biopsy sample of a female adult patient with glioblastoma multiforme. The line has now undergone over 60 passages and has been successfully cultured after cryopreservation. Immunofluorescence analyses with a panel of monoclonal antibodies were positive for glial fibrillary acidic protein and vimentin, and negative for neurofilament, galactocerebroside, and fibronectin, a pattern typical of glial cells. Based on a tetraploid, the composite karyotype of GT9 cells included the loss of chromosome 10, gain of chromosome 7, and the presence of double minute chromosomes, three of the most common karyotypic abnormalities in glioblastoma. Sequence analysis of p53 cDNA revealed a homozygous double mutation at codon 249 (commonly mutated in aflatoxin-associated hepatocellular carcinoma) and codon 250. Moreover, there was a complete absence of wild-type p53. However, unlike the majority of human glioblastomas previously described, the expression of platelet-derived growth factor-B (PDGF-B), a potent mitogenic autocrine factor, was low in GT9 cells. The expression and phosphorylation of c-Jun and Jun-B, downstream mediators of the PDGF pathway, were also low. Thus, deregulation of the PDGF pathway does not appear to be involved in the pathogenesis of the GT9 glioblastoma. Conversely, Jun-D, a negative regulator of cell growth, was also low. In addition, Phosphorylated Egr-1, a recently reported suppressor of PDGF-B/v-sis-transformed cells, was also low, suggesting that the lack of activation of the PDGF pathway was not due to these suppressive mechanisms.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: San Diego Regional Cancer Center, California 92121, USA.
authors.forename: R A
authors.initials: RA
authors.lastname: Gjerset
authors.forename: H
authors.initials: H
authors.lastname: Fakhrai
authors.forename: D L
authors.initials: DL
authors.lastname: Shawler
authors.forename: S
authors.initials: S
authors.lastname: Turla
authors.forename: O
authors.initials: O
authors.lastname: Dorigo
authors.forename: A
authors.initials: A
authors.lastname: Grover-Bardwick
authors.forename: D
authors.initials: D
authors.lastname: Mercola
authors.forename: S F
authors.initials: SF
authors.lastname: Wen
authors.forename: H
authors.initials: H
authors.lastname: Collins
authors.forename: H
authors.initials: H
authors.lastname: Lin
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D002869:Chromosome Aberrations; D002879:Chromosomes, Human, Pair 10; D002897:Chromosomes, Human, Pair 7; D004268:DNA-Binding Proteins / Q000378:metabolism; D051766:Early Growth Response Protein 1; D005260:Female; D005455:Fluorescent Antibody Technique; D016158:Genes, p53; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis; D005909:Glioblastoma / Q000235:genetics* / Q000473:pathology; D006801:Humans; D017874:Immediate-Early Proteins; D008875:Middle Aged; D008969:Molecular Sequence Data; D009154:Mutation; D010982:Platelet-Derived Growth Factor / Q000378:metabolism*; D016755:Proto-Oncogene Proteins c-jun / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000235:genetics*; D014746:Vimentin / Q000032:analysis
chemical_list: D000911:Antibodies, Monoclonal; D004268:DNA-Binding Proteins; C497325:EGR1 protein, human; D051766:Early Growth Response Protein 1; D005904:Glial Fibrillary Acidic Protein; D017874:Immediate-Early Proteins; D010982:Platelet-Derived Growth Factor; D016755:Proto-Oncogene Proteins c-jun; D014157:Transcription Factors; D016159:Tumor Suppressor Protein p53; D014746:Vimentin