Xin Jiajia, Zhang Huijie, Yin Dandan, An Ning, Chen Yaozhen, Xu Jinmei, Zhang Jing, Liu Zhixin, Liu Yongsheng, Yin Wen, Li Mingkai, Hu Xingbin
Department of Blood Transfusion, Xijing Hospital, Air Force Medical University, Xi'an, Shaanxi 710032, China.
Key Laboratory of Biochemistry and Molecular Pharmacology of Chongqing, Chongqing Medical University, Chongqing 400016, China.
Genes Dis. 2022 Sep 8;10(2):596-607. doi: 10.1016/j.gendis.2022.08.021. eCollection 2023 Mar.
With the development of tyrosine kinase inhibitor (TKI) resistance, finding the novel effective chemotherapeutic agent is of seminal importance for chronic myelogenous leukemia (CML) treatment. This study aims to find the effective anti-leukemic candidates and investigate the possible underlying mechanism. We synthesized the novel coumarin derivatives and evaluated their anti-leukemic activity. Cell viability assay revealed that compound DBH2 exhibited the potent inhibitory activity on the proliferation of CML K562 cells and TKI resistant K562 cells. Morphological observation and flow cytometry confirmed that DBH2 could selectively induce cell apoptosis and cell cycle arrest at G2/M phase of the K562 cells, which was further confirmed on the bone marrow cells from CML transgenic model mice and CD34 bone marrow leukemic cells from CML patients. Treatments of DBH2 in combination with imatinib could prolong the survival rate of SCL-tTA-BCR/ABL transgenic model mice significantly. Quantitative RT-PCR revealed that DBH2 inhibited the expression of STAT3 and STAT5 in K562 cells, and knockout alleviated the DBH2 induced apoptosis. Furthermore, DBH2 could induce the expression of PARP1 and ROCK1 in K562 cells, which may play the important role in caspase-dependent apoptosis. Our results concluded that coumarin derivative DBH2 serves as a promising candidate for the CML treatment, especially in the combination with imatinib for the TKI resistant CML, and STAT/caspase-3 pathway was involved in the molecular mechanism of anti-leukemic activity of DBH2.
随着酪氨酸激酶抑制剂（TKI）耐药性的发展，寻找新型有效的化疗药物对于慢性粒细胞白血病（CML）的治疗至关重要。本研究旨在寻找有效的抗白血病候选药物并探究其可能的潜在机制。我们合成了新型香豆素衍生物并评估了它们的抗白血病活性。细胞活力测定显示化合物DBH2对CML K562细胞和TKI耐药的K562细胞的增殖具有强效抑制活性。形态学观察和流式细胞术证实DBH2可选择性诱导K562细胞凋亡并使细胞周期停滞于G2/M期，这在CML转基因模型小鼠的骨髓细胞以及CML患者的CD34+骨髓白血病细胞中得到进一步证实。DBH2与伊马替尼联合治疗可显著延长SCL-tTA-BCR/ABL转基因模型小鼠的存活率。定量RT-PCR显示DBH2抑制K562细胞中STAT3和STAT5的表达，而敲除可减轻DBH2诱导的凋亡。此外，DBH2可诱导K562细胞中PARP1和ROCK1的表达，这可能在半胱天冬酶依赖性凋亡中起重要作用。我们的结果表明，香豆素衍生物DBH2有望成为CML治疗的候选药物，尤其是与伊马替尼联合用于TKI耐药的CML，并且STAT/半胱天冬酶-3途径参与了DBH2抗白血病活性的分子机制。
