Highlights
- •Ketoconazole induces apoptosis in HCC cells by triggering excessive mitophagy.
- •Downregulation of cyclooxygenase 2 (COX-2) is key in the induction of excessive mitophagy via the PINK1/Parkin axis.
- •HCC characterized by high COX-2 expression may benefit more from ketoconazole treatment than other subtypes.
- •Ketoconazole acts synergistically with sorafenib in the suppression of HCC growth in vitro and in vivo.
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.
Graphical abstract
Graphical Abstract
Keywords
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Article info
Publication history
Published online: October 01, 2018
Accepted:
September 21,
2018
Received in revised form:
August 28,
2018
Received:
April 4,
2018
Identification
Copyright
© 2018 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.
