Wang Xiaoning, Zhang Mei
Department of Hematology, The First Affiliated Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China.
Mol Med Rep. 2015 Jul;12(1):1449-56. doi: 10.3892/mmr.2015.3519. Epub 2015 Mar 20.
The aim of the present study was to investigate the synergistic effects of valproic acid (VPA) and arsenic trioxide (ATO) on the proliferation of RPMI8226 cells and the possible underlying mechanisms. Cell apoptosis was assessed by flow cytometry. The mRNA expression levels were analyzed by semi-quantitative polymerase chain reaction, and the protein expression levels were analyzed by western blotting. The histone acetylation and methylation states of the gene promoters were detected using a chromatin immunoprecipitation technique. The apoptotic rates of the RPMI8226 cells in the combined drug groups were significantly increased compared with those of the single drug groups (P<0.05). The mRNA and protein expression levels of Bcl-2 and the expression levels of HDAC1 mRNA and H3K9me2 protein decreased significantly in the combined groups compared with the single drug groups. The mRNA and protein expression levels of Bax, Caspase 8, Caspase 9 and LSD1, and the protein expression of acetylated H3 increased significantly in the combination groups compared with the single drug groups. Histone methylation and acetylation of the Bcl-2 and bax gene promoters were increased in the combination groups compared with the single drug groups. VPA and ATO had synergistic effects on the proliferation of RPMI8226 cells, which may have been associated with the decreased expression of Bcl-2 and the increased expression levels of Bcl-2-associated X protein, Caspase 8 and Caspase 9. Therefore, the expression levels of the Bcl-2 gene family may have been regulated by the levels of gene promoter methylation and acetylation.
本研究的目的是探讨丙戊酸(VPA)和三氧化二砷(ATO)对RPMI8226细胞增殖的协同作用及其可能的潜在机制。通过流式细胞术评估细胞凋亡。采用半定量聚合酶链反应分析mRNA表达水平,采用蛋白质印迹法分析蛋白质表达水平。利用染色质免疫沉淀技术检测基因启动子的组蛋白乙酰化和甲基化状态。与单药组相比,联合用药组RPMI8226细胞的凋亡率显著升高(P<0.05)。与单药组相比,联合组中Bcl-2的mRNA和蛋白质表达水平以及HDAC1 mRNA和H3K9me2蛋白质的表达水平显著降低。与单药组相比,联合组中Bax、Caspase 8、Caspase 9和LSD1的mRNA和蛋白质表达水平以及乙酰化H3的蛋白质表达显著增加。与单药组相比,联合组中Bcl-2和bax基因启动子的组蛋白甲基化和乙酰化增加。VPA和ATO对RPMI8226细胞的增殖具有协同作用,这可能与Bcl-2表达降低以及Bcl-2相关X蛋白、Caspase 8和Caspase 9表达水平升高有关。因此,Bcl-2基因家族的表达水平可能受基因启动子甲基化和乙酰化水平的调控。