Mirk/Dyrk1B mediates G0/G1 to S phase cell cycle progression and cell survival involving MAPK/ERK signaling in human cancer cells.
Mirk/Dyrk1B mediates G0/G1 to S phase cell cycle progression and cell survival involving MAPK/ERK signaling in human cancer cells.
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Mirk/Dyrk1B 介导人类癌细胞中涉及 MAPK/ERK 信号传导的 G0/G1 至 S 期细胞周期进展和细胞存活
DOI：
10.1186/1475-2867-13-2
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发表时间：
2013-01-11
影响因子：
5.8
通讯作者：
Shi H
中科院分区：
文献类型：
--
作者：
Gao J;Zhao Y;Lv Y;Chen Y;Wei B;Tian J;Yang Z;Kong F;Pang J;Liu J;Shi H
mirk /Dyrk1B通过破坏cyclin D1的稳定性和稳定p27kip1来维持静止的人类癌细胞的生存能力，并可能受到有丝分裂激活蛋白激酶(MAPK)/细胞外信号调节激酶（ERK）信号的负调控。本研究旨在探讨Mirk/Dyrk1B参与MAPK/ERK信号通路对人类癌细胞细胞周期和存活的影响。方法采用简单回归分析western blot检测卵巢癌和非小细胞肺癌（NSCLC）细胞中Mirk/Dyrk1B表达与ERK1/2活性的相关性。通过液相色谱-串联质谱（LC-MS/MS）蛋白质组学分析，分离出与Mirk/Dyrk1B蛋白相关位点的Mirk/Dyrk1B独特磷酸肽，并进行定量分析。用小干扰rna （sirna）和/或MEK抑制剂U0126处理人类癌细胞，持续一定时间，然后分别用流式细胞术和Western blot检测细胞周期和凋亡的变化以及相关蛋白的变化。结果在人肿瘤细胞中广泛表达的Mirk/Dyrk1B蛋白与ERK1/2的激活水平呈正相关。此外，与酪氨酸自磷酸化水平一致的Mirk/Dyrk1B蛋白表达在人类癌细胞中通过U0126或生长因子缺失培养而增加。相反，siRNA敲低Mirk/Dyrk1B导致c-Raf-MEK-ERK1/2通路激活上调，细胞周期蛋白（cyclin D1, p27kip1）随之改变，细胞生长速率增加，细胞从G0/G1期进入细胞周期S期，U0126可以剂量依赖的方式阻断，表明Mirk/Dyrk1B可隔离MAPK/ERK通路，反之亦然。而Mirk siRNA与U0126联合作用可诱导人肿瘤细胞凋亡。结论Mirk/Dyrk1B通过与MAPK/ERK信号相互作用介导细胞周期和存活，同时抑制这两条通路可能是人类癌症治疗的新靶点。
BackgroundMirk/Dyrk1B contributes to G0 arrest by destabilization of cyclin D1 and stabilization of p27kip1 to maintain the viability of quiescent human cancer cells, and it could be negatively regulated by mitogenic-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) signaling. This study was performed to investigate the effect of Mirk/Dyrk1B on cell cycle and survival of human cancer cells involving MAPK/ERK signaling.MethodsThe correlations between Mirk/Dyrk1B expression and active ERK1/2 detected by western blot in both ovarian cancer and non-small cell lung cancer (NSCLC) cells were analyzed by simple regression. Mirk/Dyrk1B unique phosphopeptides with sites associated with Mirk/Dyrk1B protein were isolated and quantitated by liquid chromatography coupled to tandem mass/mass spectrometry (LC-MS/MS) proteomics analysis. The human cancer cells were treated with small interfering RNAs (siRNAs) and/or U0126, an inhibitor of MEK for indicated duration, followed by investigating the alterations of cell cycle and apoptosis as well as related proteins examined by flow cytometry and Western blot, respectively.ResultsOur study demonstrated the widely expressed Mirk/Dyrk1B proteins in the human cancer cells were positively correlated with the levels of activated ERK1/2. Moreover, Mirk/Dyrk1B protein expressions consistent with the tyrosine autophosphorylated levels in the human cancer cells were increased by U0126 or growth factor-depleted culture. Conversely, knockdown of Mirk/Dyrk1B by siRNA led to up-regulated activation of c-Raf-MEK-ERK1/2 pathway and subsequent changes in cell cycle proteins (cyclin D1, p27kip1), accompanied by increased growth rate and cells from G0/G1 into S of cell cycle which could be blocked by U0126 in a dose-dependent manner, indicating Mirk/Dyrk1B may sequester MAPK/ERK pathway, and vice versa. Whereas, combined Mirk siRNA and U0126 induced cell apoptosis in the human cancer cells.ConclusionsThese data together show that Mirk/Dyrk1B mediates cell cycle and survival via interacting with MAPK/ERK signals and simultaneous inhibition of both pathways may be a novel therapeutic target for human cancer.
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影响因子：
4
作者：
Friedman, Eileen
通讯作者：
Friedman, Eileen
影响因子：
6.4
作者：
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通讯作者：
Friedman, Eileen
影响因子：
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作者：
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影响因子：
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作者：
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通讯作者：
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影响因子：
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作者：
Gao, JC;Niwa, K;Tamaya, T
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