Anti-fungal drug itraconazole exerts anti-cancer effects in oral squamous cell carcinoma via suppressing Hedgehog pathway
Introduction
Oral squamous cell carcinoma (OSCC) is the most frequent malignant tumor in the head and neck squamous cell carcinoma (HNSCC), with the rising overall incidence and stagnant 5-year survival rate of 50% around the world [1]. For the past 20 years, the 5-year overall survival of OSCC only increased slightly by 5% [2]. Late diagnosis, aggressive local invasion, and early lymph node metastasis result in high rates of relapse and mortality [3]. The treatment of OSCC remains a clinical challenge, despite improvements in the standard treatments, which are surgery, radiotherapy, and chemotherapy [4]. Cisplatin (DDP) is the first-line chemotherapy for OSCC with satisfactory efficacy but severe side effects [5]. Cetuximab, targeting the epidermal growth factor receptor (EGFR), was approved for the treatment of OSCC and remains the only molecular targeted therapy available for OSCC [6]. Unfortunately, long-term cetuximab treatment usually causes drug resistance over time [7,8]. Novel therapeutic strategies for OSCC are still in high demand.
Itraconazole is a broad-spectrum anti-fungal drug that blocks the synthesis of ergosterol in the fungal cell membrane [9]. The repurposing of approved drugs as novel anti-cancer agents is becoming increasingly attractive as it saves great amount of time and cost during the drug developing process [10]. Itraconazole is a Food and Drug Administration (FDA)-approved drug that had been tested for its safety for use in human and widely assessed as a novel anti-cancer agent [11]. The superior anti-cancer activity of itraconazole has been demonstrated in a variety of cancers [12]. Prior studies demonstrated that itraconazole is a promising antagonist of the Hedgehog (Hh) pathway as well as an anti-angiogenesis agent and verified it in basal cell carcinoma murine model [13,14]. However, the potential use of itraconazole in OSCC has not yet been examined. We herein aimed to investigate whether itraconazole exerts anti-cancer effects against OSCC and its underlying mechanism of action.
Section snippets
Cell culture and reagents
Human OSCC cell lines, UM1, SCC9, SCC15, SCC25 were cultured in Dulbecco's Modified Eagle Medium/Nutrient Mixture F-12 (DMEM/F – 12) (Gibco, New York, USA) supplemented with 10% fetal bovine serum (FBS) (Gibco, New York, USA). Cells were incubated in humidified air with 5% CO2 in a 37 °C incubator. For in vitro experiments, itraconazole isomer C (2S,4R,2′S) powder [15] (Aidea Pharma, Jiangsu, China) was dissolved in 0.05% dimethyl sulfoxide (DMSO). For in vivo usage, itraconazole isomer C oral
Itraconazole inhibits cell proliferation and induces cell apoptosis in OSCC cells
To investigate the anti-cancer effect of itraconazole on OSCC, four human OSCC cell lines representing distinct oral cancer subtypes were used. As shown in Fig. 1A–D, itraconazole showed appreciable, time-and dose-dependent inhibition on the proliferation of OSCC cells, including UM1, SCC9, SCC15, and SCC25. At the first 24 h, 0.1 μM itraconazole showed no inhibition on these four OSCC cells. While at 72 h, the inhibition rate of 10 μM itraconazole on these four cell lines reached 90.19%,
Discussion
This study provided evidence that itraconazole manifested significant inhibition of OSCC by negative regulation of the Hedgehog pathway. In vitro, itraconazole inhibited cell proliferation, induced cell cycle arrest and apoptosis, and perturbed cell invasion and migration. In vivo, we found that itraconazole impeded tumor growth, triggered necrosis and apoptosis in the OSCC PDX model. To our knowledge, this is the first study demonstrating the feasibility of re-purposing anti-fungal drug
Conclusion
In summary, our study demonstrated that itraconazole inhibits OSCC markedly in vitro and in vivo. The results might provide insight into the application of repurposing itraconazole in further preclinical and clinical studies for OSCC.
The following are the supplementary data related to this article.
Acknowledgments
This study was supported by the National Natural Science Foundation of China (NSFC81672659, NSFC81472523); the National Science and Technology Major Project for “Major New Drugs Innovation and Development” (2018ZX09734003); and the Natural Science Foundation of Guangdong Province (2020A1515010291).
Declaration of competing interest
The authors declared that there are no conflicts of interest.
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