Hypoxia Mediates Differential Response to Anti-EGFR Therapy in HNSCC Cells

Emilia Wiechec,Lisa Alexandersson,Jan-Ingvar Jönsson,Katharina Hansson,Karin Roberg

doi:10.3390/ijms18050943

Emilia Wiechec, Lisa Alexandersson + Show 3 more

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https://doi.org/10.3390/ijms18050943

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Abstract

Despite advances in the head and neck squamous cell carcinoma (HNSCC) treatment modalities, drug resistance and cancer recurrence are often reported. Hypoxia signaling through hypoxia-inducible factor 1 (HIF-1) promotes angiogenesis and metastasis by inducing epithelial-mesenchymal-transition (EMT). The aim of this study was to evaluate the impact of hypoxia on response to therapy as well as EMT and expression of stem cell markers in HNSCC cells. Five HNSCC cell lines (UT-SCC-2, UT-SCC-14, LK0412, LK0827, and LK0923) were selected for this study. The treatment sensitivity for radiation, cisplatin, cetuximab, and dasatinib was assessed by crystal violet assay. Gene expression of EMT and cancer stem cell (CSC) markers as well as protein level of EGFR signaling molecules were analyzed by qPCR and western blotting, respectively. Unlike UT-SCC-14 and LK0827, the LK0412 cell line became significantly more sensitive to cetuximab in hypoxic conditions. This cetuximab sensitivity was efficiently reversed after suppression of HIF-1α with siRNA. Additionally, hypoxia-induced EMT and expression of stem cell markers in HNSCC cells was partially revoked by treatment with cetuximab or knockdown of HIF-1α. In summary, our study shows that hypoxia might have a positive influence on the anti-EGFR therapy effectiveness in HNSCC. However, due to heterogeneity of HNSCC lesions, targeting HIF-1α may not be sufficient to mediate such a response. Further studies identifying a trait of hypoxia-specific response to cetuximab in HNSCC are advisable.

Highlights

Head and Neck Squamous Cell Carcinoma (HNSCC) is the sixth most common cancer worldwide, with over half a million new cases annually
We have previously screened a large number of HNSCC cell lines to determine their treatment sensitivities and phenotypes [12,13,14,15]
To investigate the effect of hypoxia on the treatment sensitivity, the HNSCC cells were exposed to cisplatin (0.1, 0.25, and 0.5 μg/mL), cetuximab (15, 30, and 60 nM), dasatinib (5, 10, and 20 nM), or ionizing γ-irradiation (2, 4, and 6 Gy)

Summary

Introduction

Head and Neck Squamous Cell Carcinoma (HNSCC) is the sixth most common cancer worldwide, with over half a million new cases annually. The modern treatment is based on a multimodality strategy involving mainly surgery and radiotherapy. Additional treatments such as cisplatin-based chemotherapy or molecular targeted drugs like cetuximab have been developed during recent years, but are only provided to patients with advanced disease. Hypoxia has been shown to negatively influence the treatment outcome in many solid tumors, and this is the case in HNSCC [1]. It arises in some areas of the tumor due to rapid tumor growth, deficient angiogenesis, vascular disturbances, and metabolic changes [2]

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