Li Peng, Zhao Qing-Li, Rehman Mati Ur, Jawaid Paras, Cui Zheng-Guo, Ahmed Kanwal, Kondo Takashi, Saitoh Jun-Ichi, Noguchi Kyo
Department of Radiology, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, Toyama 930‑0194, Japan.
Department of Environmental Health, University of Fukui School of Medical Sciences, Fukui 910‑8505, Japan.
Mol Med Rep. 2023 Feb;27(2). doi: 10.3892/mmr.2023.12928. Epub 2023 Jan 5.
The cell‑killing potential of most chemotherapeutic agents is enhanced by a temperature elevation. Isofraxidin (IF) is a coumarin compound widely found in plants, such as the Umbelliferae or Chloranthaceae families. IF induces anticancer effects in lung and colorectal cancer. To the best of our knowledge, the combined effects of hyperthermia (HT) and IF on heat‑induced apoptosis have not been reported. Acute monocytic leukemia U937 cells were exposed to HT with or without IF pre‑treatment. Apoptosis was measured by Annexin V‑FITC/PI double staining assay using flow cytometry and cell viability was observed by cell counting kit assay, DNA fragmentation. The mechanism involved in the combination was explored by measuring changes in the mitochondrial membrane potential, (MMP), intracellular ROS generation, expression of apoptosis related protein, and intracellular calcium ion level. It was demonstrated that IF enhanced HT‑induced apoptosis in U937 cells. The results demonstrated that combined treatment enhanced mitochondrial membrane potential loss and transient superoxide generation increased protein expression levels of caspase‑3, caspase‑8 and phosphorylated‑JNK and intracellular calcium levels. Moreover, the role of caspases and JNK was confirmed using a pan caspase inhibitor (zVAD‑FMK) and JNK inhibitor (SP600125) in U937 cells. Collectively, the data demonstrated that IF enhanced HT‑induced apoptosis via a reactive oxygen species mediated mitochondria/caspase‑dependent pathway in U937 cells.
大多数化疗药物的细胞杀伤潜力会因温度升高而增强。异秦皮啶（IF）是一种香豆素化合物，广泛存在于植物中，如伞形科或金粟兰科植物。IF在肺癌和结直肠癌中具有抗癌作用。据我们所知，热疗（HT）和IF对热诱导凋亡的联合作用尚未见报道。将急性单核细胞白血病U937细胞暴露于有或无IF预处理的热疗中。通过流式细胞术使用Annexin V-FITC/PI双染法测定凋亡，并通过细胞计数试剂盒测定、DNA片段化观察细胞活力。通过测量线粒体膜电位（MMP）、细胞内ROS生成、凋亡相关蛋白表达和细胞内钙离子水平的变化，探讨联合作用的机制。结果表明，IF增强了U937细胞中热疗诱导的凋亡。结果表明，联合处理增强了线粒体膜电位的丧失和瞬时超氧化物的生成，增加了caspase-3、caspase-8和磷酸化-JNK的蛋白表达水平以及细胞内钙离子水平。此外，在U937细胞中使用泛半胱天冬酶抑制剂（zVAD-FMK）和JNK抑制剂（SP600125）证实了半胱天冬酶和JNK的作用。总的来说，数据表明IF通过活性氧介导的线粒体/半胱天冬酶依赖性途径增强了U937细胞中热疗诱导的凋亡。
