Evaluation of the effects of histone deacetylase inhibitors on cells from canine cancer cell lines
Evaluation of the effects of histone deacetylase inhibitors on cells from canine cancer cell lines
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DOI：
10.2460/ajvr.69.7.938
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发表时间：
2008-07-01
影响因子：
1
通讯作者：
Chen, Ching-Shih
中科院分区：
文献类型：
--
作者：
Kisseberth, William C.;Murahari, Sridhar;Chen, Ching-Shih
目的-确定犬癌细胞暴露于组蛋白脱乙酰酶 (HDAC) 抑制剂 S(+)-N-羟基-4-(3-甲基-2-苯基-丁酰氨基)苯甲酰胺 (OSU-HDAC42) 或辛二酰苯胺异羟肟酸 (SAHA) 是否会导致组蛋白乙酰化增加和细胞活力降低，以及活力的任何变化是否涉及诱导细胞凋亡或细胞周期进展的改变。样本 Population-9 犬癌症程序-用二甲基亚砜载体、OSU-HDAC42或SARA处理来自9种犬癌细胞系的细胞，然后进行细胞活力测定。通过使用蛋白质印迹分析来评估组蛋白乙酰化。通过 ELISA 检测细胞质核小体 DNA 的碎片，并通过蛋白质印迹分析检测 caspase 3 的裂解。通过使用碘化丙啶染色和流式细胞术进行细胞周期分析。结果-分别在 6 和 4 种犬癌细胞系的细胞中达到了实现 50% 细胞活力抑制 (IC50) 所需的 OSU-HDAC42 和 SAHA 浓度，并且范围OSU-HDAC42约为0.4至1.3μM，SAHA约为0.6至4.8μM。来自 T 细胞淋巴瘤、肥大细胞瘤、骨肉瘤和组织细胞肉瘤系的细胞对 HDAC 抑制最敏感，OSU-HDAC42 的 IC(50) 小于 1pM，SAHA 的 IC(50) 小于 5 μM。通过 caspase 3 的裂解以及细胞质核小体和 subG(1) 细胞群的增加表明细胞凋亡的诱导。结论和临床相关性 - HDAC 抑制剂 OSU-HDAC42 和 SAHA 的微摩尔浓度可诱导犬癌细胞的组蛋白乙酰化、细胞毒性和细胞凋亡。一般来说，OSU-HDAC42 比 SAHA 更有效。
Objective-To determine whether exposure of canine cancer cells to histone deacetylase (HDAC) inhibitors S(+)-N-hydroxy-4-(3-methyl-2-phenyl-butyrylamino)benzamide (OSU-HDAC42) or suberoylanilide hydroxamic acid (SAHA) results in increased histone acetylation and decreased cell viability and whether any changes in viability involve induction of apoptosis or alterations in progression of the cell cycle.Sample Population-9 canine cancer cell lines.Procedures-Cells from 9 canine cancer cell lines were treated with dimethyl sulfoxide vehicle, OSU-HDAC42, or SARA, then assays of cell viability were performed. Histone acetylation was assessed by use of western blot analysis. Apoptosis was assessed via ELISA to detect fragmentation of cytoplasmic nucleosomal DNA and western blot analysis to detect cleavage of caspase 3. Cell cycle analysis was performed by use of propidium iodide staining and flow cytometry.Results-Concentrations of OSU-HDAC42 and SAHA required to achieve 50% inhibition of cell viability (IC50) were reached in cells of 6 and 4 canine cancer cell lines, respectively, and ranged from approximately 0.4 to 1.3 mu M for OSU-HDAC42 and 0.6 to 4.8 mu M for SAHA. Cells from T-cell lymphoma, mast cell tumor, osteosarcoma, and histiocytic sarcoma lines were most sensitive to HDAC inhibition, with IC(50)s of < 1pM for OSU-HDAC42 and < 5 mu M for SAHA. Induction of apoptosis was indicated via cleavage of caspase 3 and increases in cytoplasmic nucleosomes and the subG(1) cell population.Conclusions and Clinical Relevance-Micromolar concentrations of HDAC inhibitors OSU-HDAC42 and SAHA induced histone acetylation, cytotoxicity, and apoptosis in canine cancer cells. In general, OSU-HDAC42 was more potent than SAHA.