Wang Tian-yun, Jia Yan-long, Zhang Xi, Sun Qiu-li, Li Yi-chun, Zhang Jun-he, Zhao Chun-peng, Wang Xiao-yin, Wang Li
Department of Biochemistry and Molecular Biology, Xinxiang Medical University, Xinxiang 453003, Henan, China.
Henan Collaborative Innovation Center of Molecular Diagnosis and Laboratory Medicine, Xinxiang 453003, China.
Sci Rep. 2015 Dec 17;5:18443. doi: 10.1038/srep18443.
The therapeutic value of FK228 as a cancer treatment option is well known, and various types of cancer have been shown to respond to this drug. However, the complete mechanism of FK228 and the affect it has on histone lysine acetylation and the colon cancer cell proteome are largely unknown. In the present study, we used stable isotope labeling by amino acids in cell culture (SILAC) and affinity enrichment followed by high-resolution liquid chromatograph-mass spectrometer (LC-MS)/MS analysis to quantitate the changes in the lysine acetylome in HCT-8 cells after FK228 treatment. A total of 1,194 lysine acetylation sites in 751 proteins were quantified, with 115 of the sites in 85 proteins being significantly upregulated and 38 of the sites in 32 proteins being significantly downregulated in response to FK228 treatment. Interestingly, 47 histone lysine acetylation sites were identified in the core histone proteins. We also found a novel lysine acetylation site on H2BK121. These significantly altered proteins are involved in multiple biological functions as well as a myriad of metabolic and enzyme-regulated pathways. Taken together, the link between FK228 function and the downstream changes in the HCT-8 cell proteome observed in response to FK228 treatment is established.
FK228作为一种癌症治疗选择的治疗价值是众所周知的，并且已证明各种类型的癌症对这种药物有反应。然而，FK228的完整机制以及它对组蛋白赖氨酸乙酰化和结肠癌细胞蛋白质组的影响在很大程度上尚不清楚。在本研究中，我们使用细胞培养中氨基酸的稳定同位素标记（SILAC）和亲和富集，然后进行高分辨率液相色谱 - 质谱仪（LC-MS）/ MS分析，以定量FK228处理后HCT-8细胞中赖氨酸乙酰化组的变化。共定量了751种蛋白质中的1194个赖氨酸乙酰化位点，其中85种蛋白质中的115个位点在FK228处理后显著上调，32种蛋白质中的38个位点显著下调。有趣的是，在核心组蛋白中鉴定出47个组蛋白赖氨酸乙酰化位点。我们还在H2BK121上发现了一个新的赖氨酸乙酰化位点。这些显著改变的蛋白质参与多种生物学功能以及无数的代谢和酶调节途径。综上所述，建立了FK228功能与在FK228处理后观察到的HCT-8细胞蛋白质组下游变化之间的联系。
