Global Effects of DDX3 Inhibition on Cell Cycle Regulation Identified by a Combined Phosphoproteomics and Single Cell Tracking Approach.
Global Effects of DDX3 Inhibition on Cell Cycle Regulation Identified by a Combined Phosphoproteomics and Single Cell Tracking Approach.
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DOI：
10.1016/j.tranon.2018.04.001
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发表时间：
2018-06
影响因子：
5
通讯作者：
Raman V
中科院分区：
文献类型：
--
作者：
Heerma van Voss MR;Kammers K;Vesuna F;Brilliant J;Bergman Y;Tantravedi S;Wu X;Cole RN;Holland A;van Diest PJ;Raman V
DDX3 是一种具有致癌特性的 RNA 解旋酶。小分子抑制剂 RK-33 被设计成适合 DDX3 的 ATP 结合间隙，从而阻断其活性。 RK-33 在临床前癌症模型中显示出强大的活性。然而，RK-33 抗肿瘤活性背后的机制仍然很大程度上未知。在本研究中，我们使用双磷酸蛋白质组学和单细胞追踪方法来评估 RK-33 对癌细胞的作用。用 RK-33 或载体对照处理 MDA-MB-435 细胞 24 小时。使用同量异位质量标签（串联质量标签）通过定量质谱分析磷酸肽丰度的变化。在蛋白质组水平上，我们主要观察线粒体翻译、细胞分裂途径和与细胞周期进程相关的蛋白质的变化。磷酸化蛋白质组分析表明 RK-33 处理后 CDK1 活性降低。为了进一步评估 DDX3 抑制随时间推移对细胞周期进展的影响，我们在 RK-33 或 siDDX3 暴露后对荧光泛素细胞周期指示器标记的细胞进行了延时显微镜检查。单细胞追踪表明，DDX3 抑制导致细胞周期进程在间期和有丝分裂中整体延迟。此外，我们观察到核内重复的增加。总的来说，我们得出结论，DDX3 抑制会影响所有阶段的细胞，并导致整体细胞周期进展延迟。
DDX3 is an RNA helicase with oncogenic properties. The small molecule inhibitor RK-33 is designed to fit into the ATP binding cleft of DDX3 and hereby block its activity. RK-33 has shown potent activity in preclinical cancer models. However, the mechanism behind the antineoplastic activity of RK-33 remains largely unknown. In this study we used a dual phosphoproteomic and single cell tracking approach to evaluate the effect of RK-33 on cancer cells. MDA-MB-435 cells were treated for 24 hours with RK-33 or vehicle control. Changes in phosphopeptide abundance were analyzed with quantitative mass spectrometry using isobaric mass tags (Tandem Mass Tags). At the proteome level we mainly observed changes in mitochondrial translation, cell division pathways and proteins related to cell cycle progression. Analysis of the phosphoproteome indicated decreased CDK1 activity after RK-33 treatment. To further evaluate the effect of DDX3 inhibition on cell cycle progression over time, we performed timelapse microscopy of Fluorescent Ubiquitin Cell Cycle Indicators labeled cells after RK-33 or siDDX3 exposure. Single cell tracking indicated that DDX3 inhibition resulted in a global delay in cell cycle progression in interphase and mitosis. In addition, we observed an increase in endoreduplication. Overall, we conclude that DDX3 inhibition affects cells in all phases and causes a global cell cycle progression delay.
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影响因子：
82.9
作者：
通讯作者：
--
DOI：
10.1093/bioinformatics/btp026
发表时间：
2009-03-01
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Lachmann A;Ma'ayan A
通讯作者：
Ma'ayan A
DOI：
10.1073/pnas.1106245108
发表时间：
2011-07-19
影响因子：
11.1
作者：
Pek, Jun Wei;Kai, Toshie
通讯作者：
Kai, Toshie
影响因子：
4.6
作者：
Fox, Donald T.;Duronio, Robert J.
通讯作者：
Duronio, Robert J.
影响因子：
8
作者：
Botlagunta, M.;Vesuna, F.;Raman, V.
通讯作者：
Raman, V.