尼洛替尼联合三氧化二砷对K562细胞增殖及凋亡的影响 The Effects of Nilotinib and Arsenic Trioxide on the Proliferation and Apoptosis of K562 Cells

Abstract

目的：探讨尼洛替尼(AMN107)与三氧化二砷(ATO)是否具有协同作用，为寻找慢性髓细胞白血病治疗新方法提供理论基础。方法：体外培养K562细胞，分别加入AMN107、ATO及两者联合干预细胞生长；MTT法检测药物对K562细胞增殖的影响；流式细胞仪检测药物对K562细胞凋亡的影响；流式细胞仪检测药物对K562细胞的细胞周期的影响；RT-PCR检测各组K562细胞BCR-ABL融合基因表达水平的变化。结果：MTT结果示AMN107、ATO均能够抑制K562细胞的生长，但两者联合较单药效果更佳，差异有统计学意义(P < 0.05)。流式细胞仪检测细胞凋亡率(%)，AMN107 (10 umol/L)凋亡率为74.86 ± 1.86，ATO (5 umol/L)凋亡率为24.12 ± 2.03，两者联合可达到84.60 ± 1.80。RT-PCR检测显示ATO联合AMN107作用于K562细胞24 h、48 h、72 h后BCR/ABL融合基因的RQ比值分别为：0.51 ± 0.00、0.46 ± 0.00、0.37 ± 0.02。比两者单用BCR/ABL的比值更低。差异有统计学意义(P < 0.05)。 Objective: To explore whether Nilotinib (AMN107) and arsenic trioxide (ATO) had synergistic effect, to provide theoretical basis for the new treatment of chronic myelocytic leukemia. Methods: K562 cells were cultivated in vitro, treated with AMN107, ATO or both; Cell proliferation of K562 cells were evaluated by using a MTT assay. Cell apoptosis of K562 cells were examined by using Flow cytometry. Cell cycle of K562 cells were anlysised by using Flow cytometry. The expression levels of BCR-ABL fusion genes of K562 cells were analyzed by using PT-PCR. Results: MTT shows that both of the AMN107 and ATO can inhibit the growth of the cells of K562, the combination of the two drugs was better than single one. Flow cytometry detected the cell apoptosis (%), AMN107 (10 umol/L) was 74.86 ± 1.86, ATO (5 umol/L) was 24.12 ± 2.03, the combination were 84.60 ± 1.80. RT-PCR shows that the combination group treated with the K562 cells after 24 h, 48 h, 72 h, the RQ of BCR/ABL were: 0.51 ± 0.00, 0.46 ± 0.00, 0.37 ± 0.02. The results were better than single one (P < 0.05).