Li Dak Sum Yip Yio Chin Kenneth Li Marine Biopharmaceutical Research Center, Department of Marine Pharmacy, College of Food and Pharmaceutical Sciences, Ningbo University, Ningbo 315800, Zhejiang, China.
State Key Laboratory of Chemical Oncogenomics, Key Laboratory of Chemical Biology, Tsinghua Shenzhen International Graduate School, Shenzhen 518055, China.
Int J Mol Sci. 2020 Jul 18;21(14):5077. doi: 10.3390/ijms21145077.
Chronic myeloid leukemia (CML) is a malignant tumor caused by the abnormal proliferation of hematopoietic stem cells. Among a new series of acridone derivatives previously synthesized, it was found that the methoxybenzyl 5-nitroacridone derivative has nanomolar cytotoxicity in vitro against human chronic myelogenous leukemia K562 cells. In order to further explore the possible anti-leukemia mechanism of action of on K562 cells, a metabolomics and molecular biology study was introduced. It was thus found that most of the metabolic pathways of the G1 phase of K562 cells were affected after treatment. In addition, a concentration-dependent accumulation of cells in the G1 phase was observed by cell cycle analysis. Western blot analysis showed that significantly down-regulated the phosphorylation level of retinoblastoma-associated protein (Rb) in a concentration-dependent manner, upon 48 h treatment. In addition, induced K562 cells apoptosis, through both mitochondria-mediated and exogenous apoptotic pathways. Taken together, these results indicate that effectively triggers G1 cell cycle arrest and induces cell apoptosis in K562 cells, by inhibiting the CDK4/6-mediated phosphorylation of Rb. Furthermore, the possible binding interactions between and CDK4/6 protein were clarified by homology modeling and molecular docking. In order to verify the inhibitory activity of against other chronic myeloid leukemia cells, KCL-22 cells and K562 adriamycin-resistant cells (K562/ADR) were selected for the MTT assay. It is worth noting that showed significant anti-proliferative activity against these cell lines after 48 h/72 h treatment. Therefore, this study provides new mechanistic information and guidance for the development of new acridones for application in the treatment of CML.
慢性髓性白血病（CML）是一种由造血干细胞异常增殖引起的恶性肿瘤。在之前合成的一系列新的吖啶酮衍生物中，发现甲氧基苄基 5-硝基吖啶酮衍生物在体外对人慢性髓系白血病 K562 细胞具有纳摩尔级的细胞毒性。为了进一步探讨 对 K562 细胞的可能抗白血病作用机制，引入了代谢组学和分子生物学研究。结果发现， 处理后 K562 细胞的 G1 期大部分代谢途径受到影响。此外，通过细胞周期分析观察到细胞浓度依赖性地在 G1 期积累。Western blot 分析表明， 浓度依赖性地下调视网膜母细胞瘤相关蛋白（Rb）的磷酸化水平，处理 48 h 后尤为显著。此外， 通过线粒体介导和外源性凋亡途径诱导 K562 细胞凋亡。综上所述，这些结果表明， 通过抑制 CDK4/6 介导的 Rb 磷酸化， 有效触发 K562 细胞的 G1 期细胞周期停滞并诱导细胞凋亡。此外，通过同源建模和分子对接阐明了 与 CDK4/6 蛋白之间的可能结合相互作用。为了验证 对其他慢性髓性白血病细胞的抑制活性，选择 KCL-22 细胞和 K562 阿霉素耐药细胞（K562/ADR）进行 MTT 测定。值得注意的是， 处理 48 h/72 h 后，对这些细胞系表现出显著的抗增殖活性。因此，本研究为开发用于治疗 CML 的新型吖啶酮提供了新的机制信息和指导。
