State Key Laboratory of Bioreactor Engineering and Shanghai Key Laboratory of New Drug Design, School of Pharmacy, East China University of Science and Technology, Shanghai, People's Republic of China.
J Cell Biochem. 2013 Jul;114(7):1498-509. doi: 10.1002/jcb.24491.
Here we report a hydroacid named DWP0016, which exhibited HDAC inhibition and induced p53 acetylation in U251 glioblastoma cells. DWP0016 effectively inhibited the cell growth of U251 cells and other four carcinoma cell lines but did not affect the normal cells. Cell cycle distribution analysis showed DWP0016 arrested at G1 phase cell cycle dose-dependently in U251 cells. DWP0016 induced caspase-dependent and independent apoptosis in U251 cells, which was identified by flow cytometry analysis, caspases activity analysis, Western blotting assay, and caspases inhibition. Mechanisms research suggested that DWP0016 activated transcription and acetylation of tumor suppressor p53. DWP0016 regulated p300, CBP, and PCAF to facilitate p53 acetylation at lys382 in U251 cells. In addition, activation of p53 by DWP0016 promoted PUMA to catalyze mitochondrial pathway. Besides, siRNA assay indicated p53 was the key gene to induce growth inhibition, cell cycle arrest, and apoptosis in DWP0016 treated U251 cells. Conclusively, our results show DWP0016 is a potent HDAC inhibitor and the anti-tumor activity is consistent with its intended p53 activation mechanisms. These findings indicate the promising antitumor potential of DWP0016 for further glioblastoma treatment applications.
我们在此报告了一种名为 DWP0016 的水解酸，它在 U251 神经胶质瘤细胞中表现出 HDAC 抑制作用和诱导 p53 乙酰化。DWP0016 有效抑制 U251 细胞和其他四种癌细胞系的细胞生长，但不影响正常细胞。细胞周期分布分析显示，DWP0016 以剂量依赖性方式将 U251 细胞的细胞周期阻滞在 G1 期。DWP0016 诱导 U251 细胞中 caspase 依赖性和非依赖性凋亡，这通过流式细胞术分析、caspases 活性分析、Western blot 分析和 caspases 抑制得到证实。机制研究表明，DWP0016 激活了肿瘤抑制因子 p53 的转录和乙酰化。DWP0016 调节 p300、CBP 和 PCAF，以促进 U251 细胞中 p53 在赖氨酸 382 上的乙酰化。此外，DWP0016 激活 p53 促进 PUMA 催化线粒体途径。此外，siRNA 测定表明，p53 是诱导 DWP0016 处理的 U251 细胞生长抑制、细胞周期阻滞和凋亡的关键基因。总之，我们的结果表明 DWP0016 是一种有效的 HDAC 抑制剂，其抗肿瘤活性与其预期的 p53 激活机制一致。这些发现表明 DWP0016 具有进一步治疗神经胶质瘤的潜在抗肿瘤作用。
