van Oostrum I E, Rozemuller E, Knol R F, Erkens-Schulze S, Rutgers D H
Institute of Radiotherapy, University Hospital, Utrecht, The Netherlands.
Cell Tissue Kinet. 1990 Nov;23(6):523-44. doi: 10.1111/j.1365-2184.1990.tb01344.x.
Cell kinetic and histologic parameters of six xenografted tumours with volume doubling times ranging from 6 to 43 d were investigated in order to obtain kinetic information on a panel of tumours to be used in radiobiological studies. The six tumours covered a range of histologies and their DNA indices varied from 2.7 to 1.4. The length of the cell cycle (Tc), potential doubling time (Tpot) and labelling index (LI) were determined by continuous labelling with [3H]TdR and autoradiography in three tumours, Tc varied from 30 to 40 h. Determinations of the length of the S phase (Ts) were found to be less reliable by this method. Data on Ts and LI were also determined in all six tumours using bromodeoxyuridine (Brd) labelling and the single sample method: values of Tpot were slightly longer than those obtained via the autoradiographic method. In addition, multiple samples were taken after BrdU labelling. Tc was determined by fitting the data obtained from mid-S, mid-G2 and mid-G1 windows to curves described by a damped oscillator. Data obtained via the mid-S window were found to be most reliable. Generally, cell cycle times obtained by the BrdU method were longer than those observed with the autoradiographic method. Differences between the two methods could be explained by inaccuracies in the determination of Ts, LI and Tc and differences in the experimental approach. We consider the BrdU labelling method to be a suitable alternative for the time-consuming autoradiography, if data on Ts or Tpot are sufficient. Due to difficulties in the reproducibility of the immunofluorescence staining and asynchronization of cells approximately 10 h after labelling, the method of windows analysis was affected by similar problems to those observed in interpretation of percentage labelled mitosis (PLM) curves. However, the method may serve as an alternative to determine cell cycle times in vitro and, if improved technically, in vivo. Careful comparison of the data obtained from mid-S, mid-G1 and mid-G2 windows may increase the reliability of the determination of cell kinetic parameters.
为了获取一系列用于放射生物学研究的肿瘤的动力学信息，对6个异种移植肿瘤的细胞动力学和组织学参数进行了研究，这些肿瘤的体积倍增时间在6至43天之间。这6种肿瘤涵盖了多种组织学类型，其DNA指数在2.7至1.4之间。通过用[3H]TdR连续标记和放射自显影法测定了3种肿瘤的细胞周期长度（Tc）、潜在倍增时间（Tpot）和标记指数（LI），Tc在30至40小时之间。发现用这种方法测定S期（Ts）的长度不太可靠。还使用溴脱氧尿苷（Brd）标记和单样本法测定了所有6种肿瘤的Ts和LI数据：Tpot值略长于通过放射自显影法获得的值。此外，在BrdU标记后采集了多个样本。通过将从中期S、中期G2和中期G1窗口获得的数据拟合到由阻尼振荡器描述的曲线来确定Tc。发现通过中期S窗口获得的数据最可靠。一般来说，通过BrdU法获得的细胞周期时间比通过放射自显影法观察到的要长。两种方法之间的差异可以通过Ts、LI和Tc测定的不准确以及实验方法的差异来解释。如果关于Ts或Tpot的数据足够，我们认为BrdU标记法是耗时的放射自显影法的合适替代方法。由于免疫荧光染色的可重复性困难以及标记后约10小时细胞的异步化，窗口分析法受到与标记有丝分裂百分比（PLM）曲线解释中观察到类似问题的影响。然而，该方法可作为体外确定细胞周期时间的替代方法，如果在技术上得到改进，也可用于体内。仔细比较从中期S、中期G1和中期G2窗口获得的数据可能会提高细胞动力学参数测定的可靠性。
