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目的：探讨小分子化合物D609 对脑神经瘤细胞Neuro-2a 的生长抑制及诱导细胞周期阻滞的效应，并初步研究其机制。方法：采用CCK-8 法检测D609 对Neuro-2a 细胞的生长抑制作用；利用流式细胞术(FACS)检测D609 处理对细胞周期进程的影响；利用免疫印迹实验(Western blot) 检测不同浓度的D609 处理后，细胞裂解液中细胞周期蛋白抑制因子p27 的表达水平。结果： CCK-8 的实验结果显示，加入150 滋mol/L D609 处理72 小时后，细胞生长受到明显地抑制，且伴有剂量依赖效应；流式细胞术的结果表明，D609 处理使细胞周期阻滞在G0/G1 期；免疫印迹的结果表明药物处理提升了p27 的表达，且随药物浓度升高其表达亦增强。结论：D609 可以有效地抑制Neuro-2a 细胞的生长；进一步研究表明药物处理可以提升p27 的表达水平并可以诱导将细胞阻滞在G0/G1 期。因此，此研究将为脑神经瘤的治疗提供借鉴。

英文摘要:

Objective:To explore the effect of D609 on cell growth and cell cycle arrest in Neuro-2a cell line, and study the relevant mechanism.Methods:The direct effects of D609 in inhibiting the growth of Neuro-2a cells was examined by CCK-8 assay and the dose-dependent effects was also observed in this assay. Subsequently, we investigated the effects of D609 on cell cycle progression by FACS. Moreover, the expression level of p27, which is a cell-cycle regulatory protein inhibitor, was also detected by western blot when treated with various concentration of D609.Results:CCK-8 assay indicated that D609 (150 umol/L)showed significant concentration-dependent effects in inhibiting Neuro-2a cells with treatment for 72 hours. In FACS assay, D609 induced cell cycle arrest as shown bythe increased percentage of cells in G0/G1 phase. These observations were accompanied by up-regulation of cell-cycle regulatory protein inhibitorp27, which was examined by western blot.Conclusion:D609 could inhibit the growth of Neuro-2a effectively. Moreover, the drug treatment elevated the expression of p27 and further induced G0/G1 arrest. In conclusion, the research will provide useful information for treating neuroblastoma.

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