Guragain Diwakar, Seubwai Wunchana, Kobayashi Daiki, Silsinivanit Atit, Vaeteewoottacharn Kulthida, Sawanyawisuth Kanlayanee, Wongkham Chaisiri, Wongkham Sopit, Araki Norie, Cha'on Ubon
Department of Biochemistry, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
Department of Forensic Medicine, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
Cell Mol Biol (Noisy-le-grand). 2018 Jul 30;64(10):113-118.
Chemotherapy for cholangiocarcinoma (CCA) is not quite successful. In this study, we revisited the possibility of artesunate (ART) and chloroquine (CQ), the antimalarial drugs, as therapeutic agents against CCA. The possible mechanisms of these drugs to exert cytotoxicity on CCA cells were also explored. The effects of ART and CQ on proliferation and death patterns of two CCA cell lines, KKU-214 and its highly metastatic subtype KKU-214L5, were examined using water soluble tetrazolium (WST) assay and time-lapse photometry, respectively. To differentiate and verify the death patterns between necrosis and apoptosis, lactate dehydrogenase (LDH) release, and caspase 3 activity were measured. CellROXTM green reagent staining method was used to assess reactive oxygen species (ROS) production in ART- and CQ-treated cells. ART and CQ significantly inhibited proliferation of CCA cells. Both drugs kill malarial parasites via similar mechanism depending on ROS formation, however, ART induced necrotic cell death and CQ induced apoptotic cell death in CCA cells. ART induced LDH release, whereas CQ activated caspase 3, confirming induction of necrotic and apoptotic cell deaths by ART and CQ, respectively. ART treatment induced higher ROS production than CQ. ART and CQ induce CCA cells death via different death pathways. ART should be suitable for necrosis-sensitive CCA, whereas CQ is more suitable for apoptosis-sensitive CCA.
胆管癌（CCA）的化疗效果不太理想。在本研究中，我们重新探讨了抗疟药物青蒿琥酯（ART）和氯喹（CQ）作为CCA治疗药物的可能性。我们还探究了这些药物对CCA细胞产生细胞毒性的可能机制。分别使用水溶性四氮唑盐（WST）测定法和延时光度测定法，检测ART和CQ对两种CCA细胞系KKU - 214及其高转移亚型KKU - 214L5增殖和死亡模式的影响。为了区分和验证坏死与凋亡之间的死亡模式，我们检测了乳酸脱氢酶（LDH）释放和半胱天冬酶3活性。使用CellROXTM绿色试剂染色法评估ART和CQ处理的细胞中活性氧（ROS）的产生。ART和CQ显著抑制CCA细胞的增殖。两种药物通过依赖ROS形成的类似机制杀死疟原虫，然而，ART诱导CCA细胞发生坏死性细胞死亡，CQ诱导CCA细胞发生凋亡性细胞死亡。ART诱导LDH释放，而CQ激活半胱天冬酶3，分别证实了ART和CQ诱导坏死性和凋亡性细胞死亡。ART处理比CQ诱导产生更高水平的ROS。ART和CQ通过不同的死亡途径诱导CCA细胞死亡。ART适用于对坏死敏感的CCA，而CQ更适用于对凋亡敏感的CCA。
