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[通过细胞DNA含量和掺入的溴脱氧尿苷量的双变量流式细胞术测量对人脑肿瘤进行细胞动力学分析]

[Cell kinetic analysis of human brain tumors by bivariate flow cytometric measurement of cellular DNA content and amount of incorporated bromodeoxyuridine].

作者信息

Okuda Y, Taomoto K, Saya H, Ijichi A, Kudo H, Kokunai T, Tamaki N, Matsumoto S

机构信息

Department of Neurosurgery, Kobe University School of Medicine, Japan.

出版信息

No To Shinkei. 1989 Apr;41(4):383-90.

PMID:2765301
Abstract

Cell kinetics of 91 human brain tumors obtained from 88 patients were analyzed with the following two methods, 1) bivariate (two-color) flow cytometric measurement of cellular DNA content and amount of bromodeoxyuridine (BrdU) incorporated into cellular DNA, in 66 specimens, 2) immunohistochemical detection of BrdU incorporated S-phase cells, in 34 specimens. Patients were given an intravenous 1 hour infusion 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 chemically to obtain a single cell suspension, and fixed in 70% ethanol. Paraffin-embedded tissue sections were stained by the peroxidase-antiperoxidase immunohistochemical method using anti-BrdU monoclonal antibody (MoAb). Single cell suspensions were reacted with fluorescein isothiocyanate (FITC) conjugated anti-BrdU MoAb, or anti-BrdU MoAb and FITC-conjugated second antibody successively by the staining with propidium iodide, for flow cytometry (FCM). Rates of S-phase fraction in single cell suspensions 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 discrepancy 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 neuroectodermal tumors and metastatic carcinomas, moderately high in gliomas, and low in benign tumor groups.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

采用以下两种方法分析了从88例患者获取的91个人脑肿瘤的细胞动力学:1)对66个标本进行细胞DNA含量和掺入细胞DNA的溴脱氧尿苷(BrdU)量的双变量(双色)流式细胞术测量;2)对34个标本进行掺入BrdU的S期细胞的免疫组织化学检测。在手术切除前1 - 2小时,给患者静脉输注200mg/平方米的BrdU,持续1小时。切除的肿瘤标本被分成几份。一份用70%乙醇固定并石蜡包埋,另一份进行机械和/或化学消化以获得单细胞悬液,然后用70%乙醇固定。石蜡包埋的组织切片用抗BrdU单克隆抗体(MoAb)通过过氧化物酶 - 抗过氧化物酶免疫组织化学方法染色。单细胞悬液依次与异硫氰酸荧光素(FITC)偶联的抗BrdU MoAb或抗BrdU MoAb和FITC偶联的二抗反应,并用碘化丙啶染色,用于流式细胞术(FCM)。通过双变量FCM计算的单细胞悬液中的S期分数率与在组织切片中计算的标记指数(LI,即掺入BrdU的细胞百分比)相关性良好,但与Dean法分析DNA直方图的结果无关。这种差异可能是由于几个样本中的系数值较大。肿瘤的组织学恶性程度在通过双变量FCM计算的增殖指数(PI,即S + G2M期百分比)和免疫组织化学方法的LI中均有体现。PI在原始神经外胚层肿瘤和转移性癌中往往较高,在胶质瘤中中等偏高,在良性肿瘤组中较低。(摘要截短于250字)

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