Assessment of the Effect of Trichostatin A on He La Cells through FT-IR Spectroscopy
Assessment of the Effect of Trichostatin A on He La Cells through FT-IR Spectroscopy
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通过 FT-IR 光谱评估曲古抑菌素 A 对 He La 细胞的影响
DOI：
10.1021/ac504691q
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发表时间：
2015
影响因子：
7.4
通讯作者：
Li Jianxin
中科院分区：
文献类型：
--
作者：
Zhang Fengqiu;Huang Qing;Yan Jingwen;Zhang Xin;Li Jianxin
曲古抑菌素A(TSA)是一种组蛋白脱乙酰酶(HDAC)抑制剂，能抑制细胞内脱乙酰基酶的活性，促进组蛋白和非组蛋白的乙酰化。研究TSA对细胞乙酰化的影响对于更好地了解该抗肿瘤药物与癌细胞相互作用的机制是至关重要的。由于傅里叶变换红外光谱(FT-IR)是一种强大的分析工具，无需生物标记和生物标记即可无损、定量地检测生物样品，因此我们利用FT-IR光谱来探测TSA处理的细胞中蛋白质的化学和结构变化，并借助荧光显微镜观察TSA促进乙酰化水平的时间依赖和剂量效应。结果表明，TSA可引起细胞乙酰化水平的升高和细胞内蛋白质的构象/结构变化，并且随着剂量的增加，组蛋白和细胞骨架蛋白中的α-螺旋结构比例增加，乙酰化水平增加。因此，这项工作不仅验证了FT-IR光谱在细胞乙酰化定量评估中的有效性，而且可能为深入研究HDAC抑制剂药物如TSA对癌细胞的影响开辟了一条途径。
Trichostatin A (TSA) is one of histone deacetylase (HDAC) inhibitor drugs which can suppress the enzymatic activity of deacytylases and promote the acetylation of both histone and nonhistone proteins in cells. Investigation of the effect of TSA on cellular acetylation is critical for better understanding of the antitumor drug’s mechanism interacting with cancer cells. As Fourier transform infrared spectroscopy (FT-IR) is a powerful analytical tool which can detect nondestructively and quantitatively biological samples without biotagging and biolabeling, here we employed FT-IR spectroscopy to probe the chemical and structural changes of proteins in the TSA treated cells, and with the aid of fluorescent microscopy, we could scrutinize the time-dependent and dose effects on the acetylation level promoted by TSA. Our results showed that TSA caused an elevated level of cellular acetylation and conformational/structural changes of proteins in the cells, and a higher dosage of TSA caused a higher percent of α-helix structure accompanied by an increment of acetylation level in both histones and cytoskeleton proteins. This work therefore not only validates the usefulness of FT-IR spectroscopy in the quantitative assessment of cellular acetylation but also may open an avenue to the in-depth investigation of the effect of HDAC inhibitor drugs such as TSA on cancer cells.