PROTEIN KINASE CATALYTIC SUBUNIT & CELL CYCLE CHECK POINTS IN DAMAGED CELLS

多参数流式细胞术，包括BrdU含量分析， 用于确定DNA依赖性蛋白激酶 催化亚基（DNA-PKcs）在细胞周期阻滞中起作用， DNA损伤的反应。 为了研究缺乏 G1细胞周期检查点上的DNA-PKcs，DNA-PKcs +/-和-/-小鼠 胚胎成纤维细胞以6.0戈伊照射， 标记有BrdU。 照射后6小时，细胞从 DNA-PKcs +/-和-/-培养物中S期表明诱导 在G1检查站被捕 初步结果显示， 对于DNA-PKcs +/-，G1期阻滞程度相当， -/-培养物。 未来的研究将涉及连续标记， BrdU和细胞的检查在较长的时间间隔后， 辐射来检查细胞周期恢复模式。

Multiparameter flow cytometry, including BrdU content analysis, is being used to determine whether the DNA-dependent protein kinase catalytic subunit (DNA-PKcs) has a role in cell cycle arrest in response to DNA damage. To investigate the effect of the absence of DNA-PKcs on the G1 cell cycle checkpoint, DNA-PKcs +/- and -/- mouse embryonic fibroblasts were irradiated at 6.0 Gy and then continuously labeled with BrdU. By 6 h post-irradiation, cells were depleted from the S phase in both DNA-PKcs +/- and -/- cultures indicating induction of arrest at a G1 checkpoint. Preliminary results show that the extent of G1 arrest was comparable for the DNA-PKcs +/- and -/-cultures. Future studies will involve continuous labeling with BrdU and examination of the cells at longer time intervals following radiation to examine the cell cycle recovery patterns.