de Ruijter Annemieke J M, Kemp Stephan, Kramer Gertjan, Meinsma Rutger J, Kaufmann Judith O, Caron Huib N, van Kuilenburg André B P
Laboratory Genetic Metabolic Diseases, Department of Paediatrics/Emma Children's Hospital and Clinical Chemistry, Academic Medical Centre, University of Amsterdam, P.O. Box 22700, 1100 DE Amsterdam, The Netherlands.
Biochem Pharmacol. 2004 Oct 1;68(7):1279-88. doi: 10.1016/j.bcp.2004.05.010.
Neuroblastoma is a childhood cancer arising from the sympathetic nervous system. Disseminated neuroblastoma has a poor prognosis despite intensive multimodality treatment. Histone deacetylases (HDACs) were recently discovered as a potential target for pharmacological gene therapy in cancer. HDACs have an important function in regulating DNA packaging in chromatin, thereby affecting the transcription of genes. In this paper, we tested the efficacy of a newly developed histone deacetylase inhibitor, BL1521, on neuroblastoma in vitro by investigating the changes in: acetylation of histone H3, in situ HDAC activity, p21(WAF1/CIP1) and MYCN expression, metabolic activity, proliferation, morphology and the amount of apoptosis present. BL1521 inhibited the in situ HDAC activity of a panel of neuroblastoma cell lines by at least 85%. Western analysis showed an increase of histone H3 acetylation in neuroblastoma cells after incubation with BL1521. Northern analysis showed an increase in the expression of p21(WAF1/CIP1) and a decrease in the expression of MYCN in neuroblastoma cells after incubation with BL1521. Proliferation as well as the metabolic activity of neuroblastoma cells decreased significantly in response to treatment with BL1521, regardless of the MYCN status of the cells. BL1521 induced poly-(ADP-ribose) polymerase cleavage in a time- and dose-dependent manner, indicating the induction of apoptosis. Furthermore, when compared to the HDAC inhibitors Trichostatin A and 4-phenylbutyrate, BL1521 has an intermediate efficacy. Our results show that BL1521 is a potent inhibitor of HDAC and that HDACs are an attractive target for selective chemotherapy in neuroblastoma.
神经母细胞瘤是一种起源于交感神经系统的儿童癌症。尽管采用了强化多模式治疗，但播散性神经母细胞瘤的预后仍然很差。组蛋白脱乙酰酶（HDACs）最近被发现是癌症药理基因治疗的一个潜在靶点。HDACs在调节染色质中的DNA包装方面具有重要功能，从而影响基因的转录。在本文中，我们通过研究组蛋白H3的乙酰化、原位HDAC活性、p21（WAF1/CIP1）和MYCN表达、代谢活性、增殖、形态以及凋亡量的变化，测试了一种新开发的组蛋白脱乙酰酶抑制剂BL1521在体外对神经母细胞瘤的疗效。BL1521可将一组神经母细胞瘤细胞系的原位HDAC活性至少抑制85%。蛋白质免疫印迹分析显示，用BL1521孵育后，神经母细胞瘤细胞中组蛋白H3的乙酰化增加。Northern印迹分析显示，用BL1521孵育后，神经母细胞瘤细胞中p21（WAF1/CIP1）的表达增加，MYCN的表达减少。无论细胞的MYCN状态如何，用BL1521处理后，神经母细胞瘤细胞的增殖和代谢活性均显著降低。BL1521以时间和剂量依赖性方式诱导聚（ADP-核糖）聚合酶裂解，表明诱导了凋亡。此外，与HDAC抑制剂曲古抑菌素A和4-苯基丁酸相比，BL1521具有中等疗效。我们的结果表明，BL1521是一种有效的HDAC抑制剂，HDACs是神经母细胞瘤选择性化疗的一个有吸引力的靶点。
