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目的:研究组蛋白去乙酰化酶(HDAC)抑制剂丙戊酸钠(VPA)诱导髓系白血病细胞分化相关的分子标志物。方法:采用流式细胞术检测细胞表面分化抗原的表达,采用实时定量PCR和蛋白印迹法检测细胞周期相关蛋白、凋亡相关蛋白、活性氧簇(ROS)相关蛋白及组蛋白甲基化调控蛋白的表达。结果:VPA较低剂量(1 mmol/L)即诱导HL-60细胞分化,K562细胞对VPA诱导分化不敏感。VPA诱导细胞分化作用与组蛋白乙酰化基础水平及药物处理后调变间无显著相关性,VPA诱导分化与细胞周期相关蛋白、凋亡相关蛋白、ROS相关调控蛋白的表达无影响。VPA诱导分化效应与细胞不同HDAC表达水平相关,K562细胞高表达HDAC6,对VPA诱导分化不敏感,且经VPA处理后细胞组蛋白K9甲基化水平较HL-60细胞明显提高。结论:VPA诱导白血病细胞分化的作用可能与细胞HDAC6的低水平表达及组蛋白K9甲基化水平调控有关,而与组蛋白乙酰化基础水平及细胞周期相关蛋白、ROS相关调控蛋白等无关。
AIM: To investigate the molecular markers related to the differentiation of myeloid leukemia cells induced by sodium valproate (VPA), a histone deacetylase (HDAC) inhibitor. Methods: The expression of cell surface differentiation antigen was detected by flow cytometry. The expression of cell cycle related proteins, apoptosis related proteins, reactive oxygen species (ROS) related proteins and histone methylation regulatory proteins were detected by real - time PCR and Western blotting. expression. Results: HL-60 cells were induced by low dose of VPA (1 mmol / L), and K562 cells were not sensitive to VPA. There was no significant correlation between the VPA-induced cell differentiation and the basic level of histone acetylation and the post-treatment changes. VPA induced differentiation had no effect on the expression of cell cycle related proteins, apoptosis related proteins and ROS related regulatory proteins. VPA-induced differentiation was correlated with the expression of HDAC in different cell lines. K562 cells highly expressed HDAC6, which was insensitive to VPA-induced differentiation. The methylation level of K962 cells was significantly higher than that of HL-60 cells after VPA treatment. CONCLUSION: VPA-induced leukemic cell differentiation may be related to the low expression of HDAC6 and the regulation of histone K9 methylation level, but not to the basic histone acetylation level, cell cycle related proteins and ROS-related regulatory proteins.