Department of Biotherapy, State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, and West China School of Basic Medical Sciences & Forensic Medicine, Sichuan University, and Collaborative Innovation Center for Biotherapy, Chengdu 610041, PR China.
Department of Gastrointestinal Surgery, State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, and Collaborative Innovation Center for Biotherapy, Chengdu 610041, PR China.
J Hepatol. 2019 Jan;70(1):66-77. doi: 10.1016/j.jhep.2018.09.022. Epub 2018 Oct 1.
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.
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.
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.
Our results demonstrate a novel link between ketoconazole and mitophagy machinery, providing preclinical proof of concept for the use of ketoconazole in HCC treatment.
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.
肝细胞癌（HCC）是一种常见的全球癌症，仍然是一个主要的临床挑战。酮康唑，一种传统的抗真菌药物，作为癌症治疗的一种治疗选择引起了相当大的关注。然而，其作用机制尚不清楚。我们旨在评估酮康唑对 HCC 的影响，并研究其潜在机制。
我们研究了酮康唑对 HCC 细胞、细胞系衍生的异种移植物和患者来源的异种移植（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来量化酮康唑诱导的线粒体自噬。我们使用线粒体自噬抑制剂来研究线粒体自噬在酮康唑诱导的 HCC 细胞死亡中的作用。使用增益和失活方法评估环氧化酶-2（COX-2[编码为 PTGS2]）对酮康唑诱导的线粒体自噬的作用。在体内和体外测量酮康唑与索拉非尼对 HCC 的协同作用。
酮康唑在体外和体内通过触发线粒体自噬刺激 HCC 细胞凋亡。在机制上，酮康唑下调 COX-2，导致 PINK1 积累和随后 Parkin（PRKN）的线粒体易位，从而促进线粒体功能障碍介导的线粒体自噬。抑制线粒体自噬减轻了酮康唑诱导的线粒体功能障碍和细胞凋亡，支持线粒体自噬在酮康唑抗肿瘤作用中的因果关系。在 HCC PDX 模型中，酮康唑表现出明显的抗肿瘤作用，其特征为 COX-2 下调、线粒体自噬激活和凋亡诱导。此外，酮康唑与索拉非尼在体内协同抑制 HCC 异种移植物生长。
我们的研究结果表明，酮康唑与线粒体自噬机制之间存在新的联系，为酮康唑在 HCC 治疗中的应用提供了临床前概念验证。
