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The need for dynamic methods for measuring cell cycle perturbations: a study in radiation‐treated lymphoblastoid cell lines of varying p53 status

机译:对测量细胞周期扰动的动态方法的需求:辐射治疗的不同p53状态的淋巴母细胞样细胞系的研究

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摘要

Reports on the p53‐related cell cycle and apoptotic responses of EBV‐transformed lymphoblastoid cell lines to DNA damage have led to some confusion. This may be due to differences in the nature of the specific mutations under examination, but it can also be partly attributed to methodological and analytical problems (e.g. the inappropriate use of static DNA histograms for cell cycle analysis). Taking seven lymphoblastoid cell lines derived from both normal individuals and Li‐Fraumeni Syndrome/Li‐Fraumeni‐Like (LFS/LFL) patients of differing p53 status, we completed a detailed study of radiation‐induced cell cycle perturbations. Using BrdUrd pulse labelling and flow cytometry it was found that, regardless of p53 status, the cells did not arrest in G despite all of the lines showing p53 upregulation 3 hours postirradiation. The irradiated cells did, however, show a general slowing both in S‐phase entry from G and in movement through S‐phase. These facts would not have been apparent from the analysis of static DNA histograms. The problems with the use of static methods to assess changes in the dynamics of cell cycle progression apply not only to studies involving EBV‐transformed cell lines, but also to a wide range of investigations into the molecular control of cell proliferation.
机译:有关p53相关细胞周期和EBV转化的淋巴母细胞样细胞系对DNA损伤的凋亡反应的报道引起了一些混乱。这可能是由于所检查的特定突变的性质不同造成的,但也可以部分归因于方法和分析问题(例如,不适当地使用静态DNA直方图进行细胞周期分析)。收集了来自正常人和p53状态不同的Li-Fraumeni综合征/ Li-Fraumeni-Like(LFS / LFL)患者的7种淋巴母细胞细胞系,我们完成了对辐射诱导的细胞周期扰动的详细研究。使用BrdUrd脉冲标记和流式细胞仪发现,无论p53处于何种状态,尽管所有细胞系在照射后3小时都显示p53上调,但这些细胞并未停滞在G中。但是,受辐照的细胞确实显示出从G进入S相以及通过S相移动的总体速度。从静态DNA直方图的分析来看,这些事实是不明显的。使用静态方法评估细胞周期进程动态变化的问题不仅适用于涉及EBV转化细胞系的研究,而且还广泛应用于细胞增殖的分子控制研究。

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