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CELL KINETIC ANALYSIS OF HUMAN BRAIN TUMORS BY BIVARIATE FLOWCYTOMETRIC MEASUREMENT OF CELLULAR DNA CONTENT AND AMOUNT OF INCORPORATED BROMODEOXYURIDINE Yasuhiro Okuda 1 , Katsushi Taomoto 2 , Hideyuki Saya 1 , Akihiro Ijichi 1 , Hiroshi Kudo 1 , Takashi Kokunai 1 , Norihiko Tamaki 1 , Satoshi Matsumoto 1 1Department of Neurosurgery, Kobe University School of Medicine 2Department of Neurosurgery, Hyogo Prefectural Medical Center for Adults pp.383-390
Published Date 1989/4/1
DOI https://doi.org/10.11477/mf.1406206296
  • Abstract
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Cell kinetics of 91 human brain tumors obtained from 88 patients were analyzed with the following two methods, 1) bivariate (two-color) flowcyto-metric measurement of cellular DNA content and amount of bromodeoxyuridine (BrdU) incorporated into cellular DNA, in 66 specimens, 2) immunohis-tochemical detection of BrdU incorporated S-phase cells, in 34 specimens.

Patients were given an intravenous 1 hour in-fusion of 200 mg/sq. m. of BrdU 1-2 hours before the surgical removal. The excised tumor specimen was divided into several portions. One was fixed with 70% ethanol and embedded in paraffin, and another was digested mechanically and/or chemi-cally to obtian a single cell suspension, and fixed in 70% ethanol. Paraffin-embedded tissue sections were stained by the peroxidase-antiperoxidase im-munohistochemical method using anti-BrdU mono-clonal antibody (MoAb). Single cell suspensions were reacted with fluorescein isothiocyanate (FITC) conjugated anti-BrdU MoAb, or anti-BrdU MoAb and FITC-conjugated second antibody suc-cessively by the staining with propidium iodide, for flowcytometry (FCM).

Rates of S-phase fraction in single cell suspen-sions calculated by bivariate FCM were correlated well with labeling indexes (LI, i.e. the percentage of BrdU incorporated cells) calculated in tissue sections, but not with the result of analysis of DNA histogram by Dean's method. This discrepan-cy is probably due to large coefficient value in several samples. Histological malignancy of the tumors was reflected both in the proliferating index (PI, i. e. % S+G2M phase) calculated by bivariate FCM and the LI by immunohistochemical method. PI tended to be high in primitive neuro-ectodermal tumors and metastatic carcinomas, moderately high in gliomas, and low in benign tumor groups. LI in immunohistochemical studies showed the same tendency.

The advantages of bivariate FCM method were 1) able to simultaneously detect each phase (G0+ Gl, S and G2M) of the cell cycle, and polyploidy, 2) provide accurate information on over 10000 individual cells in a short time. On the other hand, immunohistochemical staining provided informa-tion about the in situ relationship between the histological malignancy and the location of BrdU incorporated cell clusters. These results indicate that the bivariate FCM analysis of tumor cell kine-tics may be useful in estimating the biological malignancy, especially in combination with the immunohistochemical staining of BrdU incorpo-rated cells.


Copyright © 1989, Igaku-Shoin Ltd. All rights reserved.

基本情報

電子版ISSN 2185-405X 印刷版ISSN 0006-8969 医学書院

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