Begg A C, Moonen L, Hofland I, Dessing M, Bartelink H
Division of Experimental Therapy, The Netherlands Cancer Institute, (Antoni van Leeuwenhoekhuis), Amsterdam.
Radiother Oncol. 1988 Apr;11(4):337-47. doi: 10.1016/0167-8140(88)90205-8.
Cell kinetic parameters in human tumours were determined by in vivo labelling with iododeoxyuridine (IUdR) followed by flow cytometric analysis of tumour biopsies after staining with a monoclonal antibody against IUdR-DNA. The purpose of this study was to determine the variation in these kinetic parameters from area to area within the same tumour. Each patient received a single i.v. injection of IUdR and the biopsy or operation specimen was taken several hours later. Multiple biopsies were taken or the operation specimen was cut into several pieces. Tumour material was stored in ethanol. Each piece was subsequently processed and stained for analysis separately. The duration of DNA synthesis (Ts), the labelling index (L.I., percent IUdR-labelled cells) and the potential doubling time (Tpot) were determined for each sample. The mean and standard deviation (variation between pieces) for each parameter was calculated for each tumour. The coefficient of variation (C.V.) provided the measure of intratumoural variation. Thirteen tumours were investigated, 6 of which were transitional cell carcinomas of the bladder and 7 of which were squamous cell carcinomas, mostly of the head and neck. Ts values ranged from 4.1 to 17.2 h (mean 9.5 h), L.I. values from 1.6 to 18.6% (mean 9.7%) and Tpot values from 2.3 to 15.1 days (mean 7.2 days). Mean C.V.s for Ts, L.I. and Tpot were 10, 24 and 27%, respectively. Most of the variation in Tpot (calculated from the other two parameters), came from the L.I., with Ts showing much less intratumoural variations. It is concluded that this kinetic method using low IUdR doses can be successfully applied in human tumours and has sufficient accuracy for predictive assay applications in which tumours need to be classified according to their proliferation rates. Further developments are required to distinguish normal and malignant cells flow cytometrically, particularly for diploid tumours.
通过用碘脱氧尿苷（IUdR）进行体内标记确定人类肿瘤中的细胞动力学参数，随后用抗IUdR-DNA单克隆抗体染色后对肿瘤活检组织进行流式细胞术分析。本研究的目的是确定同一肿瘤内不同区域这些动力学参数的变化。每位患者静脉注射一次IUdR，数小时后采集活检标本或手术标本。采集多个活检标本或把手术标本切成数块。肿瘤组织保存在乙醇中。随后将每一块组织分别进行处理和染色以进行分析。测定每个样本的DNA合成持续时间（Ts）、标记指数（L.I.，IUdR标记细胞的百分比）和潜在倍增时间（Tpot）。计算每个肿瘤每个参数的平均值和标准差（各块之间的差异）。变异系数（C.V.）提供了肿瘤内变异的度量。研究了13个肿瘤，其中6个是膀胱移行细胞癌，7个是鳞状细胞癌，大多位于头颈部。Ts值范围为4.1至17.2小时（平均9.5小时），L.I.值范围为1.6至18.6%（平均9.7%），Tpot值范围为2.3至15.1天（平均7.2天）。Ts、L.I.和Tpot的平均C.V.分别为10%、24%和27%。Tpot的大部分变异（根据其他两个参数计算）来自L.I.，而Ts显示出的肿瘤内变异要少得多。得出结论，这种使用低剂量IUdR的动力学方法可成功应用于人类肿瘤，并且对于需要根据增殖率对肿瘤进行分类的预测性检测应用具有足够的准确性。需要进一步改进以便通过流式细胞术区分正常细胞和恶性细胞，特别是对于二倍体肿瘤。
