Lai Kuan-Ming, Tseng Ruo-Han, Shih Yu-Hung, Huang Ying-Chih
Program in Translational Medicine, National Chung Hsing University, Taichung, Taiwan, R.O.C.
Hemato-Oncology Division Department of Internal Medicine, Changhua Christian Hospital, Changhua, Taiwan, R.O.C.
Anticancer Res. 2024 Mar;44(3):1033-1044. doi: 10.21873/anticanres.16898.
BACKGROUND/AIM: Chemotherapy drugs for leukemia, such as 5-azacytidine (Aza), have often various adverse effects. Hesperetin (Hes), a naturally occurring compound, is a potential adjuvant agent for anticancer therapy. This study aimed to investigate the effect of an Aza-Hes combination on acute leukemia cell lines, which elucidates the role of combination treatment in leukemia progression.
HL-60 and U937 cells were treated with Aza and Hes at various concentrations or their combination. Cell proliferation and apoptosis was evaluated using the Cell Counting Kit-8 assay and annexin V/propidium iodide staining, respectively. Cell cycle analysis was conducted using flow cytometry. The expression of apoptosis-related and cell cycle-related proteins in leukemia cells was analyzed through western blotting. The synergistic effect of the Aza and Hes agents was estimated using the Chou-Talalay method.
We observed that Aza or Hes monotherapy engendered a dose-dependent reduction in HL-60 and U937 cell viability. However, treatment with the Aza-Hes combination for 24 h synergistically inhibited U937 cell proliferation by inducing both apoptosis and S-phase cell cycle arrest. Furthermore, the Aza-Hes combination down-regulated p-ERK and p-c-Jun N-terminal kinase expression and up-regulated p-p38 expression.
Overall, our findings indicate that the Aza-Hes combination induces apoptosis and S-phase cell-cycle arrest through the mitogen-activated protein kinase pathway. In conclusion, the Aza-Hes combination is a potential antileukemia treatment.
背景/目的：白血病化疗药物，如5-氮杂胞苷（Aza），常有多种不良反应。橙皮素（Hes）是一种天然存在的化合物，是抗癌治疗的潜在辅助剂。本研究旨在探讨Aza-Hes联合用药对急性白血病细胞系的影响，以阐明联合治疗在白血病进展中的作用。
用不同浓度的Aza和Hes或其组合处理HL-60和U937细胞。分别使用细胞计数试剂盒-8法和膜联蛋白V/碘化丙啶染色评估细胞增殖和凋亡。采用流式细胞术进行细胞周期分析。通过蛋白质印迹分析白血病细胞中凋亡相关和细胞周期相关蛋白的表达。使用Chou-Talalay法评估Aza和Hes药物的协同作用。
我们观察到Aza或Hes单药治疗均导致HL-60和U937细胞活力呈剂量依赖性降低。然而，Aza-Hes联合用药24小时通过诱导凋亡和S期细胞周期阻滞协同抑制U937细胞增殖。此外，Aza-Hes联合用药下调p-ERK和p-c-Jun N端激酶表达，上调p-p38表达。
总体而言，我们的研究结果表明，Aza-Hes联合用药通过丝裂原活化蛋白激酶途径诱导凋亡和S期细胞周期阻滞。总之，Aza-Hes联合用药是一种潜在的抗白血病治疗方法。
