Department of Biological Science and Technology, China Medical University, Taichung, Taiwan.
Department of Food Nutrition and Health Biotechnology, Asia University, Taichung, Taiwan.
Environ Toxicol. 2021 May;36(5):764-772. doi: 10.1002/tox.23078. Epub 2020 Dec 21.
Genistein (GEN) has been shown to induce apoptotic cell death in various human cancer cells. L-asparaginase (Asp), a clinical drug for leukemia, has been shown to induce cell apoptosis in leukemia cells. No available information concerning GEN combined with Asp increased the cell apoptosis compared to GEN or Asp treatment alone. The objective of this study is to evaluate the anti-leukemia activity of GEN combined with Asp on human leukemia HL-60 cells in vitro. The cell viability, the distribution of cell cycle, apoptotic cell death, and the level of ΔΨ were examined by flow cytometric assay. The expressions of apoptosis-associated proteins were measured by western blotting. GEN combined with Asp revealed a more significant decrease in total viable cells and induced a higher percentage of G2/M phase arrest, DNA damage, and cell apoptosis than that of GEN or Asp treatment only in HL-60 cells. Furthermore, the combined treatments (GEN and Asp) showed a higher decrease in the level of ΔΨ than that of GEN or Asp treatment only. These results indicated that GEN combined with Asp induced mitochondria dysfunction by disrupting the mitochondrial membrane potential. The results from western blotting demonstrated that the treatment of GEN combined with Asp showed a higher increase in the levels of Bax and Bak (pro-apoptotic proteins) and an active form of caspase-3 and a higher decrease in Bcl-2 (anti-apoptotic protein) than that of GEN or Asp treatment alone. GEN significantly enhances the efficiency of Asp on cytotoxic effects (the induction of apoptosis) in HL-60 cells.
染料木黄酮（GEN）已被证明可诱导多种人类癌细胞的凋亡细胞死亡。L-天冬酰胺酶（Asp）是一种用于白血病的临床药物，已被证明可诱导白血病细胞凋亡。目前尚无关于 GEN 与 Asp 联合使用相对于 GEN 或 Asp 单独治疗可增加细胞凋亡的信息。本研究旨在评估 GEN 与 Asp 联合用于体外人白血病 HL-60 细胞的抗白血病活性。通过流式细胞术检测细胞活力、细胞周期分布、凋亡细胞死亡和ΔΨ水平。通过 Western blot 测量凋亡相关蛋白的表达。与 GEN 或 Asp 单独处理相比，GEN 与 Asp 联合使用可更显著降低总活细胞，并诱导更高比例的 G2/M 期阻滞、DNA 损伤和细胞凋亡。此外，联合治疗（GEN 和 Asp）显示ΔΨ水平下降幅度高于 GEN 或 Asp 单独治疗。这些结果表明，GEN 与 Asp 联合使用通过破坏线粒体膜电位诱导线粒体功能障碍。Western blot 的结果表明，与 GEN 或 Asp 单独处理相比，GEN 与 Asp 联合处理显示 Bax 和 Bak（促凋亡蛋白）水平升高和 caspase-3 活性形式增加，Bcl-2（抗凋亡蛋白）水平降低。GEN 显著提高了 Asp 在 HL-60 细胞中的细胞毒性作用（诱导凋亡）的效率。
