title: Proliferation and cell death of human glioblastoma cells after carbon-ion beam exposure: morphologic and morphometric analyses.
abstract: Histological analyses of glioblastoma cells after carbon-ion exposure are still limited and ultrastructural characteristics have not been investigated in detail. Here we report the results of morphological and morphometric analyses of a human glioblastoma cell line, CGNH-89, after ionizing radiation to characterize the effect of a carbon-beam on glioblastoma cells. Using CGNH-89 cells exposed to 0-10 Gy of X-ray (140 kVp) or carbon-ions (18.3 MeV/nucleon, LET=108 keV/microm), we performed conventional histology and immunocytochemistry with MIB-1 antibody, transmission electron microscopy, and computer-assisted, nuclear size measurements. CGNH-89 cells with a G to A transition in codon 280 in exon 8 of the TP53 gene had nuclei with pleomorphism, marked nuclear atypia and brisk mitotic activity. After carbon-ion and X-ray exposure, living cells showed decreased cell number, nuclear condensation, increased atypical mitotic figures, and a tendency of cytoplasmic enlargement at the level of light microscopy. The deviation of the nuclear area size increased during 48 h after irradiation, while the small cell fraction increased in 336 h. In glioblastoma cells of the control, 5 Gy carbon-beam, and 10 Gy carbon-beam, and MIB-1 labeling index decreased in 24 h (12%, 11%, 7%, respectively) but increased in 48 h (10%, 20%, 21%, respectively). Ultrastructurally, cellular enlargement seemed to depend on vacuolation, swelling of mitochondria, and increase of cellular organelles, such as the cytoskeleton and secondary lysosome. We could not observe apoptotic bodies in the CGNH-89 cells under any conditions. We conclude that carbon-ion irradiation induced cell death and senescence in a glioblastoma cell line with mutant TP53. Our results indicated that the increase of large cells with enlarged and bizarre nuclei, swollen mitochondria, and secondary lysosome occurred in glioblastoma cells after carbon-beam exposure.
authors.affiliation: Department of Human Pathology, Gunma University Graduate School of Medicine, Maebashi, Gunma, Japan.
authors.forename: Takuma
authors.initials: T
authors.lastname: Oishi
authors.forename: Atsushi
authors.initials: A
authors.lastname: Sasaki
authors.forename: Nobuyuki
authors.initials: N
authors.lastname: Hamada
authors.forename: Shogo
authors.initials: S
authors.lastname: Ishiuchi
authors.forename: Tomoo
authors.initials: T
authors.lastname: Funayama
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Sakashita
authors.forename: Yasuhiko
authors.initials: Y
authors.lastname: Kobayashi
authors.forename: Takashi
authors.initials: T
authors.lastname: Nakano
authors.forename: Yoichi
authors.initials: Y
authors.lastname: Nakazato
mesh_terms: D002244:Carbon; D016923:Cell Death / Q000528:radiation effects*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000528:radiation effects; D016158:Genes, p53 / Q000528:radiation effects; D005909:Glioblastoma / Q000378:metabolism / Q000648:ultrastructure*; D020450:Heavy Ions; D006801:Humans; D007150:Immunohistochemistry; D019394:Ki-67 Antigen / Q000528:radiation effects; D018499:Linear Energy Transfer; D046529:Microscopy, Electron, Transmission; D009154:Mutation
chemical_list: D019394:Ki-67 Antigen; D002244:Carbon