Yang Ching-Huey, Tung Kuo-Lung, Wu Yen-Ting, Liu Cheng, Lin Sheng-Chieh, Yang Chun-Chuan, Wu Chin-Han, Chang Hong-Yi, Wu Shih-Yi, Huang Bu-Miin, Lan Yu-Yan
Department of Traditional Chinese Medicine, Kaohsiung Veterans General Hospital, Pingtung Branch, Pingtung 91245, Taiwan.
Department of Dental Technology, Shu-Zen Junior College of Medicine and Management, Kaohsiung 82144, Taiwan.
Evid Based Complement Alternat Med. 2021 Nov 2;2021:9925684. doi: 10.1155/2021/9925684. eCollection 2021.
Since a portion of patients with nasopharyngeal carcinoma (NPC) do not benefit much from current standard treatments, it is still needed to discover new therapeutic drugs to improve the prognosis of the patients. Considering that Chinese traditional medicine plays a role in inhibiting tumor progression, in this study, we aimed to investigate whether a Chinese herbal formula, Qing Yan Li Ge Tang (QYLGT), has the anticancer activity in NPC cells and explore the underlying mechanism as well. MTT assay, colony formation assay, immunoblotting assay, and DNA laddering assay were performed to assess cell viability, cell colony formation, protein expression, and DNA fragmentation, respectively. Results show that QYLGT was able to inhibit the cell viability and decrease colony formation ability in NPC cells. QYLGT could also increase the formation of intracellular vacuoles and induce the autophagy-related protein expressions, including Atg3, Atg6, and Atg12-Atg5 conjugate in NPC cells. Treatment with an autophagy inhibitor, 3-methyladenine, could significantly recover QYLGT-inhibited cell viability of NPC cells. In addition, QYLGT did not significantly induce apoptosis in NPC cells. We also found that QYLGT had the ability to activate phosphoinositide 3-kinase (PI3K)/Akt/mammalian target of the rapamycin (mTOR) pathway. Treatment with PI3K inhibitors, LY294002 and wortmannin, or mTOR inhibitors, rapamycin and Torin 1, could not only recover QYLGT-inhibited cell viability of NPC cells but also inhibit Atg3 expression. Taken together, our results demonstrated that QYLGT could induce autophagic cell death in NPC cells through the PI3K/Akt/mTOR pathway.
由于一部分鼻咽癌（NPC）患者从当前的标准治疗中获益不多，因此仍需要发现新的治疗药物来改善患者的预后。考虑到中药在抑制肿瘤进展中发挥作用，在本研究中，我们旨在研究中药方剂清咽利膈汤（QYLGT）是否对NPC细胞具有抗癌活性，并探索其潜在机制。分别进行MTT法、集落形成试验、免疫印迹试验和DNA梯状条带试验来评估细胞活力、细胞集落形成、蛋白质表达和DNA片段化。结果表明，QYLGT能够抑制NPC细胞的活力并降低其集落形成能力。QYLGT还可增加NPC细胞内空泡的形成并诱导自噬相关蛋白的表达，包括Atg3、Atg6以及Atg12-Atg5偶联物。用自噬抑制剂3-甲基腺嘌呤处理可显著恢复QYLGT抑制的NPC细胞活力。此外，QYLGT未显著诱导NPC细胞凋亡。我们还发现QYLGT具有激活磷酸肌醇3-激酶（PI3K）/蛋白激酶B（Akt）/雷帕霉素哺乳动物靶蛋白（mTOR）通路的能力。用PI3K抑制剂LY294002和渥曼青霉素或mTOR抑制剂雷帕霉素和Torin 1处理，不仅可恢复QYLGT抑制的NPC细胞活力，还可抑制Atg3表达。综上所述，我们的结果表明，QYLGT可通过PI3K/Akt/mTOR通路诱导NPC细胞发生自噬性细胞死亡。
