Comparative Proteomic Analysis of Histone Modifications upon Acridone Derivative 8a-induced CCRF-CEM Cells by Data Independent Acquisition
Comparative Proteomic Analysis of Histone Modifications upon Acridone Derivative 8a-induced CCRF-CEM Cells by Data Independent Acquisition
复制标题
通过数据独立采集对吖啶酮衍生物 8a 诱导的 CCRF-CEM 细胞组蛋白修饰进行比较蛋白质组学分析
DOI：
10.1021/acs.jproteome.9b00650
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发表时间：
2020-02-01
影响因子：
4.4
通讯作者：
Qin，Jun
中科院分区：
文献类型：
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作者：
Wang，Yini;Xu，Caixia;Qin，Jun
该先导化合物吖啶酮衍生物8通过与DNA碱基直接堆积并引发CCRF-CEM细胞中的ROS而诱导DNA损伤，从而显示出有效的抗增殖活性。为了明确DNA损伤感知和修复过程中的染色质改变，在QE-plus上采用数据独立采集（Data Independent Acquisition，DIA）方法对8 a影响的CCRF-CEM细胞中单个和共存的组蛋白翻译后修饰（PTM）进行了详细的定量图谱。共定量了79种不同的和164种共存的组蛋白PTM，其中16种不同的组蛋白PTM在将8a处理的细胞与载体对照细胞进行比较时显著改变。通过对三个H3和一个H4组蛋白标记物的Western印迹分析证实组蛋白PTM的变化。H3 K9、H3 K36和H4 K20的二甲基化水平上调，提示CCRF-CEM细胞可能通过加速DNA损伤修复来对抗8a诱导的DNA损伤，这一点通过53 BP 1灶在损伤DNA上的定位增加得到了证实。大多数显著改变的PTM参与转录调控，包括下调H3 K18、H3 K27和H3 K122的乙酰化，以及上调H3 K9和H3 K27的二甲基化和三甲基化。流式细胞仪检测结果显示，这种转录沉默现象与G2/M期细胞阻滞有关。本研究表明，DIA蛋白质组学策略提供了一种灵敏和准确的方法来表征CCRF-CEM细胞在8处理后共存的组蛋白PTM的变化及其相互作用。具体而言，组蛋白PTM重排转录沉默和细胞周期停滞DNA损伤修复可能有助于8a影响的表观遗传反应的机制。
The lead compound acridone derivative8ashowed potent antiproliferative activity by inducing DNA damage through direct stacking with DNA bases and triggering ROS in CCRF-CEM cells. To define the chromatin alterations during DNA damage sensing and repair, a detailed quantitative map of single and coexisting histone post-translational modifications (PTMs) in CCRF-CEM cells affected by8awas performed by the Data Independent Acquisition (DIA) method on QE-plus. A total of 79 distinct and 164 coexisting histone PTMs were quantified, of which 16 distinct histone PTMs were significantly altered when comparing8a-treated cells with vehicle control cells. The changes in histone PTMs were confirmed by Western blotting analysis for three H3 and one H4 histone markers. The up-regulated dimethylation on H3K9, H3K36, and H4K20 implied that CCRF-CEM cells might accelerate DNA damage repair to counteract the DNA lesion induced by8a, which was verified by an increment in the 53BP1 foci localization at the damaged DNA. Most of the significantly altered PTMs were involved in transcriptional regulation, including down-regulated acetylation on H3K18, H3K27, and H3K122, and up-regulated di- and trimethylation on H3K9 and H3K27. This transcription-silencing phenomenon was associated with G2/M cell cycle arrest after8atreatment by flow cytometry. This study shows that the DIA proteomics strategy provides a sensitive and accurate way to characterize the coexisting histone PTMs changes and their cross-talk in CCRF-CEM cells after8atreatment. Specifically, histone PTMs rearrange transcription-silencing, and cell cycle arrest DNA damage repair may contribute to the mechanism of epigenetic response affected by8a.