组蛋白去乙酰化酶抑制剂联合FMS样酪氨酸激酶-3抑制剂对FLT3-ITD突变白血病细胞增殖、凋亡和周期的影响

目的 探讨组蛋白去乙酰化酶抑制剂(HDACi)联合FMS样酪氨酸激酶-3(FLT3)抑制剂对FLT3-ITD突变的急性髓系白血病(AML)细胞系增殖抑制作用及其相关机制。方法 将西达本胺(Chidamide)和奎扎替尼(Quizartinib, AC220)以不同浓度单药或联合作用于MV4-11、MOLM-13细胞系48 h后，CCK8检测细胞增殖能力；Compusyn 1.0软件分析两药联合作用的协同效应；流式细胞计数法测定各组细胞凋亡率、增殖周期；蛋白质印迹法测定P53、Bcl-2、Bax、FLT3、磷酸化FLT3(p-FLT3)及磷酸化AKT(p-AKT)蛋白表达水平。结果 随西达本胺及奎扎替尼单药或联合作用浓度升高，对MV4-11及MOLM-13细胞系的48h增殖抑制率均显著升高(P<0.05)，并且两药具有协同抑制效应(均CI值<1)。在奎扎替尼1.0 nmol/L+西达本胺1.0μmol/L作用48 h后，细胞凋亡率MV4-11细胞株为31.697%±5.648%,MOLM-13细胞株为18.500%±1.751%，细胞周期阻滞于G0/G1期的比例MV4-11细...

Effects of HDACi combined with FLT3i on proliferation, apoptosis and cycle of FLT3-ITD (+) acute myelocytic leukemia cells

Objective This study was conducted to investigate whether histone deacetylase inhibitor (HDACi) combined with FMS-like tyrosine kinase 3 (FLT3) inhibitor can synergistically inhibit the proliferation of Acute myeloid leukemia (AML) cells with FLT3-In-ternal tandem duplication (ITD) and explore its related mechanisms, aiming to provide a preclinical basis for obtaining a more provendrug treatment modality for AML patients with FLT3-ITD mutation.Methods Chidamide and quizartinib were administered to MV4-11 and MOLM-13 cell lines at different concentrations alone or in combination for 48 h. The following assays were performed: CCK8 todetect cell proliferation ability.Compusyn 1.0 software was used to analyze the synergistic effect of the combination of the two drugs.Theapoptosis rate and proliferation cycle of each group were detected by flow cytomety.The expression levels of P53, Bcl-2, Bax, FLT3, p-FLT3 and p-AKT proteins, which are key downstream factors of PI3K/AKT signaling pathway were detected by Western-blotting.Re. sults Chidamide and quizartinib had proliferation inhibitory effects on MV4-11 and MOLM-13 cell lines in a dose-dependent manner. Chidamide combined with quizartinib had synergistic proliferation inhibitory effects on MV4-11 and MOLM-13 cell lines.Chidamide and quizartinib alone could promote the apoptosis and block the cell cycle of MV4-11 and MOLM-13 cell lines.Chidamide combined with quizartinib make more effects.The combination treatment with quizartinib and chidamide significantly up-regulated P53 and Bax pro-apoptotic protein, and down-regulated the expression of p-FLT3, anti-apoptotic protein Bcl-2 and p-Akt, a key downstream factor of PI3K/AKT signaling pathway, compared with single drug.Conclusion The combination of chidamide and quizartinib can promote theapoptosis and block the cell cycle of the AML cells,resulting in the suppression of the FLT3-ITD(+ ) AML cells, mainly through the downregulation of Bcl-2 and the upregulation of P53 and Bax proteins.