Ketoconazole exacerbates mitophagy to induce apoptosis by downregulating cyclooxygenase-2 in hepatocellular carcinoma
Ketoconazole exacerbates mitophagy to induce apoptosis by downregulating cyclooxygenase-2 in hepatocellular carcinoma
复制标题
酮康唑通过下调肝细胞癌中的环氧合酶 2 加剧线粒体自噬诱导细胞凋亡
DOI：
10.1016/j.jhep.2018.09.022
复制
发表时间：
2019-01-01
影响因子：
25.7
通讯作者：
Huang, Canhua
中科院分区：
文献类型：
--
作者：
Chen, Yan;Chen, Hai-Ning;Huang, Canhua
背景与目的：肝细胞癌是世界范围内常见的癌症，一直是临床上的一大难题。酮康唑是一种传统的抗真菌药物，作为癌症治疗的一种选择，已经引起了相当大的关注。然而，其作用机制仍未明确。我们的目的是评估酮康唑对肝癌细胞的作用并探讨其可能的机制。方法：我们检测了酮康唑对肝癌细胞、细胞系来源的异种移植瘤和患者来源的异种移植瘤(PDX)模型的抗肿瘤作用。通过免疫荧光、免疫印迹和透射电子显微镜分析，对酮康唑诱导的细胞有丝分裂吞噬作用进行定量。我们使用吞噬丝裂酶抑制剂来研究吞丝分裂在酮康唑诱导的肝癌细胞死亡中的作用。采用增益法和功能损失法评价环氧合酶-2(COX-2[由PTGS2编码])在酮康唑诱导的细胞有丝分裂中的作用。结果：酮康唑在体内外均通过激活有丝分裂促进肝癌细胞的凋亡。在机制上，酮康唑下调COX-2，导致PINK1积聚，继而导致Parkin(PRKN)线粒体易位，从而促进线粒体吞噬功能障碍。抑制丝裂原吞噬可以减轻酮康唑诱导的线粒体功能障碍和细胞凋亡，支持吞丝分裂在酮康唑抗肿瘤作用中的因果作用。在肝癌PDX模型中，酮康唑表现出明显的抗肿瘤作用，其特征是COX-2表达下调，丝裂原激活，并诱导细胞凋亡。此外，酮康唑与索拉非尼具有协同抑制肝癌移植瘤体内生长的作用。结论：我们的结果显示了酮康唑与有丝分裂吞噬机制之间的新联系，为酮康唑用于肝癌治疗提供了临床前的概念证据。我们的研究表明，酮康唑是一种广谱抗真菌药物，通过下调COX-2激活PINK1/Parkin介导的有丝分裂吞噬，从而加速细胞凋亡，从而抑制肝癌的生长。此外，酮康唑与索拉非尼在体内外抑制肝癌生长方面具有协同作用。(C)2018年欧洲肝脏研究协会。爱思唯尔出版，版权所有。
Background & Aims: Hepatocellular carcinoma (HCC) is a common cancer worldwide and remains a major clinical challenge. Ketoconazole, a traditional antifungal agent, has attracted considerable attention as a therapeutic option for cancer treatment. However, its mechanism of action is still not clearly defined. We aimed to evaluate the effect of ketoconazole on HCC and investigate the underlying mechanisms.Methods: We examined the antitumor effect of ketoconazole on HCC cells, cell line-derived xenografts, and a patient-derived xenograft (PDX) model. Ketoconazole-induced mitophagy was quantified by immunofluorescence, immunoblotting and transmission electron microscopy analysis. We used mitophagy inhibitors to study the role of mitophagy on HCC cell death induced by ketoconazole. The role of cyclooxygenase-2 (COX-2 [encoded by PTGS2]) on ketoconazole-induced mitophagy was evaluated using gain- and loss-of-function methods. The synergistic effect of ketoconazole with sorafenib on HCC was measured in vivo and in vitro.Results: Ketoconazole stimulated apoptosis in HCC cells by triggering mitophagy in vitro and in vivo. Mechanistically, ketoconazole downregulated COX-2, which led to PINK1 accumulation and subsequent mitochondrial translocation of Parkin (PRKN), and thereby promoted mitophagy-mediated mitochondrial dysfunction. Inhibiting mitophagy alleviated ketoconazole-induced mitochondrial dysfunction and apoptosis, supporting a causal role for mitophagy in the antitumor effect of ketoconazole. In the HCC PDX model, ketoconazole demonstrated a marked antitumor effect characterized by COX-2 downregulation, mitophagy activation, and apoptosis induction. Moreover, ketoconazole acted synergistically with sorafenib to suppress HCC xenograft growth in vivo.Conclusion: Our results demonstrate a novel link between ketoconazole and mitophagy machinery, providing preclinical proof of concept for the use of ketoconazole in HCC treatment.Lay summary: Hepatocellular carcinoma (HCC) is a common malignancy worldwide and remains a major clinical challenge. Our study reveals that ketoconazole, a broad-spectrum antifungal agent, activates PINK1/Parkin-mediated mitophagy by downregulating COX-2, consequently resulting in the acceleration of apoptosis and thereby inhibiting the growth of HCC. Furthermore, ketoconazole acts synergistically with sorafenib in the suppression of HCC growth in vitro and in vivo. (C) 2018 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.