Trichostatin A (TSA) is a natural derivative of diene isohydroxamic acids. Trichostatin A is a histone deacetylase inhibitor (IC50=1.8 nM) that is reversible and specific. Trichostatin A leads to the hyperacetylation of core histones, which regulates chromatin structure.

HeLa cells:20 nM, MCF-7 cells:500 nM, MDA-MB-231 cells:500 nM, HDAC:1.8 nM, HDAC (Vascular Smooth Muscle cells):0.08 μM, HDAC1:6.0 ± 2.5 nM, HDAC6:8.6 ± 1.4 nM, HDAC4:38 ± 4 nM

方法：八种乳腺癌细胞 MCF-7、T-47D、ZR-75-1、BT-474、MDA-MB-231、MDA-MB-453、CAL 51 和 SK-BR-3 用 Trichostatin A (10−12 -10−5 M) 处理 96 h，使用 SRB 方法检测细胞活力。
结果：Trichostatin A 抑制八种乳腺癌细胞系的增殖，平均 IC50=124.4±120.4 nM（范围 26.4-308.1 nM）。[1]
方法：食管鳞状细胞癌细胞 EC9706 和 EC1 用 Trichostatin A (0.3-1μM) 处理 48 h，使用 Flow Cytometry 方法检测细胞凋亡情况。
结果：在 0.3 和 0.5 μM Trichostatin A 的剂量下，早期凋亡的百分比没有显著增加。但与对照组相比，1.0 μM Trichostatin A 处理可显著诱导早期细胞凋亡。此外，中晚期凋亡的百分比以浓度依赖的方式增加。[2]
方法：食管鳞状细胞癌细胞 EC9706 和 EC1 用 Trichostatin A (0.3-1μM) 处理 60 min，使用 Western Blot 方法检测靶点蛋白表达水平。
结果：Trichostatin A 以剂量依赖的方式降低 PI3K 的蛋白水平以及 p-Akt 和 p-ERK1/2。组蛋白H4的乙酰化以浓度依赖性方式增加。[2]

方法：为检测体内抗肿瘤活性，将 Trichostatin A (500 μg/kg) 皮下注射给 NMU 诱导乳腺癌肿瘤的大鼠，每天一次，持续四周。
结果：Trichostatin A 在体内具有显著的抗肿瘤活性。Trichostatin A 处理的大鼠肿瘤具有良性表型，纤维腺瘤或管状腺瘤，这表明 Trichostatin A 的抗肿瘤活性可能归因于分化的诱导。[1]
方法：为检测体内抗肿瘤活性，将 Trichostatin A (0.5-1 mg/kg，每周两次) 和 Quercetin (10 mg/kg，每周三次) 腹腔注射给携带人肺腺癌肿瘤 A549 的裸鼠，持续十三周。
结果：高剂量 Trichostatin A 显著抑制肿瘤生长，而低剂量 Trichostatin A 和 Quercetin 单独使用没有效果。然而，低剂量 Trichostatin A 和 Quercetin 联合治疗显著抑制了肿瘤生长。[3]

In vitro HDAC activity: Total cellular extracts are prepared from each breast cancer cell line (MCF-7, T-47D, ZR-75-1, BT-474, MDA-MB-231, MDA-MB-453, CAL 51, or SK-BR-3). A 20 μL crude cell extract (~2.5 ×105 cells), in the presence of varying concentrations of Trichostatin A in 0.1% (v/v) ethanol or 0.1% (v/v) ethanol as vehicle control, are incubated for 60 minutes at 25 °C with 1 μL (~1.5 × 106 cpm) of [3H]acetyl-labeled histone H4 peptide substrate (NH2-terminal residues 2-20) that has been acetylated with [3H]acetic acid, sodium salt (3.7 GBq/mmol) by an in vitro incorporation method. Each 200 μL reaction is quenched with 50 μL of 1 M HCl/0.16 M acetic acid and extracted with 600 μL of ethyl acetate, and released [3H]acetate is quantified by scintillation counting. IC50 values are determined graphically using nonlinear regression to fit inhibition data to the appropriate dose-response curve.

Cells are exposed to various concentrations of Trichostatin A for 96 hours. After treatment, cell proliferation is estimated using the sulforhodamine B colorimetric assay. Cell viability is determined by trypan blue exclusion. (Only for Reference)