Nicotiana benthamiana-derived epidermal growth factor rescues human oral keratinocytes exposed to 5-fluorouracil cytotoxicity
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Abstract
Objectives: 5-fluorouracil (5-FU) is commonly used as a chemotherapy drug for head and neck cancers. However, it often produces an adverse cytotoxic effect on the oral epithelia leading to oral mucositis (OM) in cancer patients. Thus, reversing this cytotoxic effect locally with novel bioactive drugs or factors is necessary. This study aimed to investigate the rescue effect of Nicotiana benthamiana plant-derived epidermal growth factor (P-EGF) on human normal oral keratinocytes (NOKs) exposed to 5-FU-induced cytotoxicity.
Materials and Methods: Cytotoxicity and ATP-dependent proliferation and metabolism of NOK monolayer cultures were assessed after 48 hours of 5-FU exposure and 48 hours of P-EGF treatment using MTT and luciferase-based ATP assays, respectively. Cell death was determined with Live/Dead staining using calcein-AM and propidium iodide. Cell recolonization was evaluated by in vitro scratch assay. Five biological replicates were run. One-way ANOVA with post hoc Dunnett’s test was used for statistical analysis with GraphPad Prism 9.5.1 software.
Results: P-EGF at 80ng/ml significantly promoted cell proliferation and metabolism after 5-FU-induced cytotoxicity (p-value=0.0006). Additionally, the recolonization of NOKs was also significantly enhanced by the addition of P-EGF (20, 40 and 80 ng/ml, p-value<0.0001).
Conclusion(s): P-EGF rescued the proliferation of 5-FU-exposed NOKs as well as induced epithelial cell recolonization. Thus, P-EGF could be a potential therapeutic approach for OM.
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