Department of Oncology, Transplant and Advances in Medicine, Section of Hematology, University of Pisa, Ospedale S Chiara, Via Roma 67, 56126 Pisa, Italy.
Anticancer Res. 2010 Nov;30(11):4525-35.
Cytotoxic and pro-apoptotic effects exerted by the histone deacetylase inhibitor ITF2357 have been reported in acute myeloid leukemia HL-60 cells. In the current study, its mechanism of action was investigated at the molecular level.
Cell proliferation was evaluated by methyl thiazol tetrazolium bromide reduction; apoptosis by annexin V, mitochondrial transmembrane potential by tetramethylrhodamine ethyl ester. Functional experiments and gene expression evaluations were performed by flow cytometry, microarray, and quantitative polymerase chain reaction.
Significant cell growth inhibition and increased apoptosis were observed. ITF2357 reduced protein levels of BCL-2, MCL-1, and BCL-X, and increased levels of BAK. Exposure to ITF2357 did not abrogate NF-κB DNA binding. After microarray analysis, interleukin-10, interleukin-6, epidermal growth factor, peroxisome proliferator-activated receptor (PPAR), transforming growth factor β, P38 mitogen-activated protein kinase, aryl hydrocarbon receptor, xenobiotic metabolism, PPAR/retinoic acid receptor, NF-κB, apoptosis, lipopolysaccharide/interleukin-1, G-protein receptor, T-cell receptor, and platelet-derived growth factor were the de-regulated pathways.
This study shows that ITF2357 influences both proliferation and inflammatory pathways in HL-60 cells; this observation could have possible applications in clinical practice.
组蛋白去乙酰化酶抑制剂 ITF2357 在急性髓系白血病 HL-60 细胞中已被报道具有细胞毒性和促凋亡作用。在本研究中，从分子水平研究了其作用机制。
通过噻唑蓝溴化还原法评估细胞增殖；通过膜联蛋白 V 评估细胞凋亡；通过四甲基罗丹明乙酯评估线粒体跨膜电位。通过流式细胞术、微阵列和实时定量聚合酶链反应进行功能实验和基因表达评估。
观察到明显的细胞生长抑制和凋亡增加。ITF2357 降低了 BCL-2、MCL-1 和 BCL-X 的蛋白水平，增加了 BAK 的水平。ITF2357 暴露并没有消除 NF-κB 的 DNA 结合。经过微阵列分析，白细胞介素-10、白细胞介素-6、表皮生长因子、过氧化物酶体增殖物激活受体（PPAR）、转化生长因子-β、P38 丝裂原激活蛋白激酶、芳香烃受体、外来化合物代谢、PPAR/视黄酸受体、NF-κB、凋亡、脂多糖/白细胞介素-1、G 蛋白受体、T 细胞受体和血小板衍生生长因子是失调的途径。
本研究表明 ITF2357 影响 HL-60 细胞的增殖和炎症途径；这一观察结果可能在临床实践中有潜在的应用。
