Duan Hong, Heckman Caroline A, Boxer Linda M
Center for Molecular Biology in Medicine, Veterans Affairs, Palo Alto Health Care System, Palo Alto, CA, USA.
Mol Cell Biol. 2005 Mar;25(5):1608-19. doi: 10.1128/MCB.25.5.1608-1619.2005.
Histone deacetylase (HDAC) inhibitors are promising antitumor agents, but they have not been extensively explored in B-cell lymphomas. Many of these lymphomas have the t(14;18) translocation, which results in increased bcl-2 expression and resistance to apoptosis. In this study, we examined the effects of two structurally different HDAC inhibitors, trichostatin A (TSA) and sodium butyrate (NaB), on the cell cycle, apoptosis, and bcl-2 expression in t(14;18) lymphoma cells. We found that in addition to potent cell cycle arrest, TSA and NaB also dramatically induced apoptosis and down-regulated bcl-2 expression, and overexpression of bcl-2 inhibited TSA-induced apoptosis. The repression of bcl-2 by TSA occurred at the transcriptional level. Western blot analysis and quantitative chromatin immunoprecipitation (ChIP) assay showed that even though HDAC inhibitors increased overall acetylation of histones, localized histone H3 deacetylation occurred at both bcl-2 promoters. TSA treatment increased the acetylation of the transcription factors Sp1 and C/EBPalpha and decreased their binding as well as the binding of CBP and HDAC2 to the bcl-2 promoters. Mutation of Sp1 and C/EBPalpha binding sites reduced the TSA-induced repression of bcl-2 promoter activity. This study provides a mechanistic rationale for the use of HDAC inhibitors in the treatment of human t(14;18) lymphomas.
组蛋白去乙酰化酶(HDAC)抑制剂是很有前景的抗肿瘤药物,但在B细胞淋巴瘤中尚未得到广泛研究。许多这类淋巴瘤存在t(14;18)易位,导致bcl-2表达增加和对凋亡产生抗性。在本研究中,我们检测了两种结构不同的HDAC抑制剂曲古抑菌素A(TSA)和丁酸钠(NaB)对t(14;18)淋巴瘤细胞的细胞周期、凋亡及bcl-2表达的影响。我们发现,除了强力的细胞周期阻滞作用外,TSA和NaB还显著诱导凋亡并下调bcl-2表达,且bcl-2的过表达抑制了TSA诱导的凋亡。TSA对bcl-2的抑制作用发生在转录水平。蛋白质印迹分析和定量染色质免疫沉淀(ChIP)试验表明,尽管HDAC抑制剂增加了组蛋白的整体乙酰化,但在两个bcl-2启动子处均发生了局部组蛋白H3去乙酰化。TSA处理增加了转录因子Sp1和C/EBPα的乙酰化,降低了它们与bcl-2启动子的结合以及CBP和HDAC2与bcl-2启动子的结合。Sp1和C/EBPα结合位点的突变降低了TSA诱导的bcl-2启动子活性抑制。本研究为HDAC抑制剂用于治疗人类t(14;18)淋巴瘤提供了机制依据。