Comprehensive analysis for histone acetylation of human colon cancer cells treated with a novel HDAC inhibitor.
Comprehensive analysis for histone acetylation of human colon cancer cells treated with a novel HDAC inhibitor.
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DOI：
10.2174/13816128113199990531
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发表时间：
2014-03
影响因子：
3.1
通讯作者：
Yunlong Zhao;X. Fang;Ye Wang;Junmei Zhang;Sheng Jiang;Zhe Liu;Zhenyi Ma;Liyan Xu;Enmin Li-Enmin
中科院分区：
文献类型：
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作者：
Yunlong Zhao;X. Fang;Ye Wang;Junmei Zhang;Sheng Jiang;Zhe Liu;Zhenyi Ma;Liyan Xu;Enmin Li-Enmin
大量证据表明，组蛋白赖氨酸乙酰化异常与癌症的发生在表观遗传学水平上密切相关。组蛋白在赖氨酸上的乙酰化主要由“铅笔”-组蛋白乙酰转移酶（HAT）和“橡皮”-组蛋白去乙酰化酶HDAC调节。整体组蛋白去乙酰化的显著升高被认为是癌症的生物标志物。因此，目前的抗肿瘤药物设计通常靶向HDAC，用天然或合成的小分子如largazole抑制肿瘤细胞中过表达的HDAC。最近，通过用酪氨酸取代瓦尔1设计并制备了一种新的largazole衍生物（largazole-7），这种修饰使其对人癌细胞的选择性比正常细胞高100倍以上。然而，目前尚不清楚在这种药物的处理下组蛋白乙酰化的动态水平。目前还不清楚其他修饰是否也受到largazole-7处理的影响。因此，对该药物作用的癌细胞系的组蛋白修饰的全局作图可能对阐明其分子机制和探索其作为抗肿瘤药物的潜力非常有益。为了实现这一目标，我们将细胞培养物中氨基酸的稳定同位素标记（SILAC）和高分辨率MS相结合，用于对人结肠癌细胞（HCT-116）的组蛋白赖氨酸乙酰化和其他修饰进行全面鉴定和定量分析，其中有和没有largazole-7处理。在这项分析中，我们在核心组蛋白的38个位点上鉴定了68个组蛋白PTM，包括赖氨酸乙酰化、甲基化和丁酰化，这是一种新的赖氨酸修饰。进一步的定量分析不仅发现乙酰化赖氨酸的总体增加，而且观察到在药物刺激下赖氨酸甲基化和丁酰化丰度的变化。据我们所知，这是第一个报告，largazole-7对赖氨酸丁酰化的调控。我们的研究扩展了癌症组蛋白标记的目录，并为了解药物治疗下已知和新的表观遗传标记提供了一种方法。
Extensive evidence suggests that dysregulation of histone lysine acetylation is intimately linked with the development of cancer in epigenetic level. Histone acetylation on lysine is regulated mainly by the "pencil"--Histone acetyltransferases (HATs) and the "eraser"--Histone deacetylases HDACs. Dramatic elevation of global histone deacetylation is considered as a biomarker for cancer. Therefore, current antitumor drug design often targets HDACs, inhibiting overexpressed HDAC in tumor cells with natural or synthesized small molecules like largazole. Recently, a novel largazole derivative (largazole-7) was designed and prepared by replacement of Val 1 with tyrosine, and this modification increases selectivity toward human cancer cells over normal cells more than 100-fold. However, it is unclear about the dynamic level of histone acetylation under the treatment of this drug. It is also unclear whether the other modifications are also affected by largazole-7 treatment. Therefore, a global mapping of modifications on the histone proteins of cancer cell line treated by this drug may be of great benefit to elucidating its molecular mechanisms and exploring its potent as an antitumor drug. To realize the goal, we combined stable isotope labeling by amino acids in cell culture (SILAC) and high resolution MS for comprehensive identification and quantitative analysis of histone lysine acetylation and other modifications of Human Colon Cancer Cells (HCT-116) with and without treatment of largazole-7. In this analysis, we identified 68 histone PTMs in 38 sites on core histones, including lysine acetylation, methylation and butyrylation, a novel lysine modification. Further quantitative analysis not only discovered the global increased acetylated lysines, but also observed the changes of abundance of lysine methylation and butyrylation under stimulation of the drug. To our knowledge, it is the first report that regulation of largazole-7 against lysine butyrylation. Our study expands the catalog of histone marks in cancer, and provides an approach for understanding the known and new epigenetic marks under treatment of drugs.