H2O2 induces a transient multi-phase cell cycle arrest in mouse fibroblasts through modulating cyclin D and p21Cip1 expression
H2O2 induces a transient multi-phase cell cycle arrest in mouse fibroblasts through modulating cyclin D and p21Cip1 expression
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DOI：
10.1074/jbc.m111123200
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发表时间：
2002-04-19
影响因子：
4.8
通讯作者：
Lam, EWF
中科院分区：
文献类型：
--
作者：
Barnouin, K;Dubuisson, ML;Lam, EWF
为了防御活性氧引起的潜在损伤，增殖细胞进入短暂的细胞周期停滞。我们用H2 O2处理小鼠成纤维细胞，发现亚致死剂量的H2 O2在G、S和G处诱导了短暂的多相细胞周期停滞。阶段，而不是M阶段。Western blot分析表明，这种短暂的细胞周期阻滞与细胞周期蛋白D1和D3的下调和CKI p21（Cip 1）表达的上调有关。我们还证明H2 O2对p21（Cip 1）表达的诱导至少部分是在转录水平介导的，并且可以在p53功能缺失的情况下发生。进一步的免疫沉淀激酶和免疫耗竭试验表明，在响应于H2 O2处理，细胞周期蛋白Ds的表达下调与细胞周期蛋白D-CDK 4的抑制有关，而P21（Cip 1）的积累负责抑制细胞周期蛋白E和A-CDK 2的活性，并与细胞周期蛋白B-CDC 2的活性下调有关。这些数据可以解释H2 O2刺激后细胞周期停滞在G（1），S和G（2）期。缺失p21（Cip 1），恢复细胞周期蛋白D的表达，或过度表达细胞周期蛋白E单独是不足以有效地克服亚致死剂量的H2 O2引起的细胞周期停滞。相比之下，人类疱疹病毒8 K细胞周期蛋白的过表达，它可以模拟细胞周期蛋白D和E的功能，足以推翻这种短暂的细胞周期停滞。我们的研究结果的基础上，我们提出了一个模型，其中适度水平的过氧化氢诱导短暂的多相细胞周期停滞至少部分通过上调P21（Cip 1）和下调细胞周期蛋白D的表达。
To defend against the potential damages induced by reactive oxygen species, proliferating cells enter a transient cell cycle arrest. We treated mouse fibroblasts with H2O2 and found that sublethal doses of H2O2 induced a transient multi-phase cell cycle arrest at the G, S, and G. phases but not the M phase. Western blot analysis demonstrated that this transient cell cycle arrest is associated with the down-regulation of cyclins D1 and D3 and up-regulation of the CKI p21(Cip1) expression. We also demonstrate that the induction in p21(Cip1) expression by H2O2 is at least partially mediated at the transcriptional level and can occur in the absence of p53 function. Further immunoprecipitation kinase and immunodepletion assays indicated that in response to H2O2 treatment, the down-regulation of cyclin Ds expression are associated with repression of cyclin D-CDK4, whereas the accumulation of P21(Cip1) is responsible for the inhibition of cyclin E and A-CDK2 activity and associated with the down-regulation of cyclin B-CDC2 activity. These data could account for the cell cycle arrest at the G(1), S, and G(2) phases following H2O2 stimulation. Deletion of p21(Cip1), restoration of cyclin D expression, or overexpression of cyclin E alone is insufficient to effectively overcome the cell cycle arrest caused by sublethal doses of H2O2. By contrast, overexpression of the human Herpesvirus 8 K cyclin, which can mimic the function of cyclin D and E, is enough to override this transient cell cycle arrest. On the basis of our findings, we propose a model in which moderate levels of H2O2 induce a transient multi-phase cell cycle arrest at least partially through up-regulation of P21(Cip1) and down-regulation of cyclin D expression.