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学位論文 |
学位
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Tanaka, Yuko ; Sasaki, Atsushi ; Ishiuchi, Shogo ; Nakazato, Yoich
概要:
application/pdf<br />Thesis or Dissertation<br />To characterize the cellular density and proliferative activity of glia
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l fibrillary acidic protein (GFAP)-negative cells in pilocytic astrocytoma (PA), surgically excised tissues of PAs (n=37) and diffuse astrocytomas (DAs) (n=11) were examined morphologically and immunohistochemically using antibodies against GFAP, Olig2, Iba1 and Ki-67 (MIB-1). In PA, Olig2 immunoreactivity was significantly expressed in protoplasmic astrocytes in microcystic, loose areas and cells in oligodendroglioma-like areas. Iba1-positive, activated microglia/macrophages were also commonly observed in microcystic areas. In compact areas, a prominent reaction for GFAP was observed, but for Olig2 and Iba1 to a lesser degree. On semiquantitative analysis, the number of Olig2-positive cells was significantly higher in PAs (mean labeling index (LI) ± standard deviation (SD): 46.8 ± 15.4%) than in DAs (13.3 ± 7.8%) (P<0.001). Many Iba1-positive, microglia/macrophages were observed in PAs (19.9 ± 6.5%), similarly to DAs (20.9 ± 9.9%). Re-immunostaining of PA demonstrated that most Ki-67-positive, proliferating cells expressed Olig2, whereas GFAP or Iba1 expression in Ki-67-positive cells was less frequent (14.7 ± 13.7%, and 8.8 ± 13.6%) in a double immunostaining study. Conversely, the percentage of Olig2-positive, proliferating cells in total Olig2-positive cells (7.2 ± 3.9%) was higher than that of Iba1-positive, proliferating cells in total Iba1-positive cells (0.9 ± 0.6%). In conclusion, the present study found that PA consisted of numerous GFAP-negative cells, including Olig2-positive cells with high proliferation. Semiquantitative analysis of Olig2 immunohistochemistry in microcystic areas might therefore be useful for the differential diagnosis of PA and DA<br />学位記番号：医博甲1088
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| 2.
学位論文 |
学位
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Oishi, Takuma ; Sasaki, Aatsushi ; Hamada, Nobuyuki ; Ishiuchi, Shogo ; Funayama, Tomoo ; Sakashita, Tetsuya ; Kobayashi, Yasuhiko ; Nakano, Takashi ; Nakazato, Yoichi
概要:
application/pdf<br />Thesis or Dissertation<br />Histological analyses of glioblastoma cells after carbon-ion exposure a
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re 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 (140kVp) or carbon-ions (18.3 MeV/nucleon, LET = 108 keV/μm), 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 hours after irradiation, while the small cell fraction increased in 336 hours. In glioblastoma cells of the control, 5 Gy carbon-beam, and 10 Gy carbon-beam, and MIB-1 labeling index decreased in 24 hours (12%, 11%, 7%, respectively) but increased in 48 hours (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.<br />学位記番号：医博甲1096
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