Abstract
The potential clinical use of ultrasound in inducing cell apoptosis and enhancing the effects of anticancer drugs in the treatment of cancers has previously been investigated. In this study, the combined effects of low-intensity ultrasound (LIU) and cetuximab, an anti-epidermal growth factor receptor (EGFR) antibody, on cell killing and induction of apoptosis in HSC-3 and HSC-4 head and neck cancer cells, and its mechanisms were investigated. Experiments were divided into 4 groups: non-treated (CNTRL), cetuximab-treated (CETU), ultrasound-treated (UST) and the combination of cetuximab and US-treated (COMB). Cell viability was assessed by trypan blue staining assay and induction of apoptosis was detected by fluorescein isothiocyanate (FITC)-Annexin V and propidium iodide (PI) staining assay at 24 h after cetuximab and/or US treatment. To elucidate the effect of cetuximab and US on EGFR signaling and apoptosis in head and neck cancer cells after the treatments, the expression of EGFR, phospho-EGFR, and the activation of caspase-3 were evaluated with western blotting. More cell killing features were evident in the COMB group in HSC-3 and HSC-4 cells compared with the other groups. No differences in EGFR expression among the CETU, UST and COMB groups was observed, while the expression of phospho-EGFR in the CETU group was downregulated compared with that in the CNTRL group. Phospho-EGFR expression was much more downregulated in the COMB group compared with that in the other groups. In addition, the activation of caspase-3 in the UST group was upregulated compared with that in the CNTRL group. Caspase-3 activation was much more upregulated in the COMB group than that in the other groups. These data indicated that LIU was able to enhance the anticancer effect of cetuximab in HSC-3 and HSC-4 head and neck cancer cells.
Highlights
Head and neck squamous cell carcinoma (HNSCC) is known for its rapid clinical progression and poor prognosis
There were no differences of the expression of epidermal growth factor receptor (EGFR) among the CETU, UST and COMB groups, while the expression of phospho-EGFR was down-regulated and the cleavage of caspase-3 was upregulated in the COMB group compared with the other groups
These outcomes include the enhancement of cellular uptake of drugs and the induction of cell killing or apoptosis [15,16,22]
Summary
Head and neck squamous cell carcinoma (HNSCC) is known for its rapid clinical progression and poor prognosis. The survival rates for many types of HNSCC have improved little over the past 40 years. Anti-monoclonal antibody is often used as a first-line and primary treatment of malignancies, such as malignant lymphomas and breast cancer. An anti-epidermal growth factor receptor (EGFR) antibody, is the only EGFR inhibitor approved in HNSCC. Several combined methods with cetuximab, such as irradiation or many types of anticancer drugs, have been applied to enhance the effect of treatment. The efficacy of such treatment is inadequate. New therapeutic methods should be developed to improve the survival rate
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