Environmental Stress Affects the Activity of Metabolic and Growth Factor Signaling Networks and Induces Autophagy Markers in MCF7 Breast Cancer Cells
Environmental Stress Affects the Activity of Metabolic and Growth Factor Signaling Networks and Induces Autophagy Markers in MCF7 Breast Cancer Cells
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DOI：
10.1074/mcp.m113.034751
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发表时间：
2014-03-01
影响因子：
7
通讯作者：
Cutillas, Pedro R.
中科院分区：
文献类型：
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作者：
Casado, Pedro;Bilanges, Benoit;Cutillas, Pedro R.
磷酸化蛋白质组学技术有助于我们理解信号通路如何相互作用和调节生物过程。该技术还被用于表征信号网络在疾病进展过程中如何重塑，并识别信号通路活性和癌症治疗反应的生物标志物。这些研究中的一个潜在警告是，磷酸化是一种非常动态的修饰，在实验或细胞样本的检索和处理过程中可能会发生很大变化。在这里，我们研究了细胞暴露于环境条件如何调节 MCF7 乳腺癌细胞系的磷酸化和信号通路活性。将细胞暴露于环境条件 15 分钟后，测量的 3,500 个位点中约 1.5% 显示磷酸化程度发生显着变化。这种扰动改变磷酸化模式的效果并不涉及由于激酶和磷酸酶的随机激活而引起的随机变化。相反，细胞暴露于环境条件下会引发环境应激反应，涉及对代谢应激情况的协调反应，其中包括：（1）AMPK 的激活； (2) PI3K、AKT、ERK的抑制； (3)翻译延伸水平上蛋白质合成抑制标记物的增加； (4)自噬标记物增加。我们还观察到，将细胞维持在冰中会改变但并没有完全消除这种代谢应激反应。总之，细胞暴露于环境条件会影响先前与代谢和生长因子信号传导有关的信号网络的活性。质谱数据已存入 ProteomeXchange，标识符为 PXD000472。
Phosphoproteomic techniques are contributing to our understanding of how signaling pathways interact and regulate biological processes. This technology is also being used to characterize how signaling networks are remodeled during disease progression and to identify biomarkers of signaling pathway activity and of responses to cancer therapy. A potential caveat in these studies is that phosphorylation is a very dynamic modification that can substantially change during the course of an experiment or the retrieval and processing of cellular samples. Here, we investigated how exposure of cells to ambient conditions modulates phosphorylation and signaling pathway activity in the MCF7 breast cancer cell line. About 1.5% of 3,500 sites measured showed a significant change in phosphorylation extent upon exposure of cells to ambient conditions for 15 min. The effects of this perturbation in modifying phosphorylation patterns did not involve random changes due to stochastic activation of kinases and phosphatases. Instead, exposure of cells to ambient conditions elicited an environmental stress reaction that involved a coordinated response to a metabolic stress situation, which included: (1) the activation of AMPK; (2) the inhibition of PI3K, AKT, and ERK; (3) an increase in markers of protein synthesis inhibition at the level of translation elongation; and (4) an increase in autophagy markers. We also observed that maintaining cells in ice modified but did not completely abolish this metabolic stress response. In summary, exposure of cells to ambient conditions affects the activity of signaling networks previously implicated in metabolic and growth factor signaling. Mass spectrometry data have been deposited to the ProteomeXchange with identifier PXD000472.