Loss of p53 induces M-phase retardation following G2 DNA damage checkpoint abrogation.
Loss of p53 induces M-phase retardation following G2 DNA damage checkpoint abrogation.
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DOI：
10.1016/s0003-9861(03)00010-9
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发表时间：
2003-04
影响因子：
3.9
通讯作者：
Y. Minemoto;Sanae Uchida;M. Ohtsubo;M. Shimura;T. Sasagawa;M. Hirata;H. Nakagama;Y. Ishizaka;K. Yamashita
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文献类型：
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作者：
Y. Minemoto;Sanae Uchida;M. Ohtsubo;M. Shimura;T. Sasagawa;M. Hirata;H. Nakagama;Y. Ishizaka;K. Yamashita
大多数缺乏功能性p53蛋白的细胞系由于DNA损伤而停滞在细胞周期的G2期。当G2检查点被废除时，这些细胞被迫进入有丝分裂灾难。A549肺腺癌细胞，其中p53与HPV 16 E6基因消除，表现出有效的阻滞在G2期时，用阿霉素处理。给G2期阻滞的细胞施用咖啡因会引起细胞表型的急剧变化，其性质取决于p53的状态。流式细胞仪和显微镜观察显示，细胞，无论是含有或缺乏p53恢复其细胞周期，并进入有丝分裂后咖啡因治疗。然而，在p53阴性的细胞中，过渡到M期比p53阳性的细胞慢。与这些观察结果一致，在p53阴性细胞中，CDK 1活性保持在高水平，沿着稳定的细胞周期蛋白B1。加入丁内酯I，这是一种CDK 1和CDK 2的抑制剂，p53阴性细胞减少了漂浮的圆形细胞群，并诱导细胞周期蛋白B1的消失。这些结果表明，p53通路和泛素介导的有丝分裂细胞周期蛋白的降解之间的关系和可能的G2-DNA损伤检查点和有丝分裂检查点之间的串扰。
Most cell lines that lack functional p53 protein are arrested in the G2 phase of the cell cycle due to DNA damage. When the G2 checkpoint is abrogated, these cells are forced into mitotic catastrophe. A549 lung adenocarcinoma cells, in which p53 was eliminated with the HPV16 E6 gene, exhibited efficient arrest in the G2 phase when treated with adriamycin. Administration of caffeine to G2-arrested cells induced a drastic change in cell phenotype, the nature of which depended on the status of p53. Flow cytometric and microscopic observations revealed that cells that either contained or lacked p53 resumed their cell cycles and entered mitosis upon caffeine treatment. However, transit to the M phase was slower in p53-negative cells than in p53-positive cells. Consistent with these observations, CDK1 activity was maintained at high levels, along with stable cyclin B1, in p53-negative cells. The addition of butyrolactone I, which is an inhibitor of CDK1 and CDK2, to the p53-negative cells reduced the floating round cell population and induced the disappearance of cyclin B1. These results suggest a relationship between the p53 pathway and the ubiquitin-mediated degradation of mitotic cyclins and possible cross-talk between the G2-DNA damage checkpoint and the mitotic checkpoint.