A Wnt-mediated phenotype switch along the epithelial–mesenchymal axis defines resistance and invasion downstream of ionising radiation in oral squamous cell carcinoma

Background Dynamic transitions of tumour cells along the epithelial–mesenchymal axis are important in tumorigenesis, metastasis and therapy resistance. Methods In this study, we have used cell lines, 3D spheroids and tumour samples in a variety of cell biological and transcriptome analyses to highlight the cellular and molecular dynamics of OSCC response to ionising radiation. Results Our study demonstrates a prominent hybrid epithelial–mesenchymal state in oral squamous cell carcinoma cells and tumour samples. We have further identified a key role for levels of E-cadherin in stratifying the hybrid cells to compartments with varying levels of radiation response and radiation-induced epithelial–mesenchymal transition. The response to radiation further entailed the generation of a new cell population with low expression levels of E-cadherin, and positive for Vimentin (ECAD Low/Neg -VIM Pos ), a phenotypic signature that showed an enhanced capacity for radiation resistance and invasion. At the molecular level, transcriptome analysis of spheroids in response to radiation showed an initial burst of misregulation within the first 30 min that further declined, although still highlighting key alterations in gene signatures. Among others, pathway analysis showed an over-representation for the Wnt signalling pathway that was further confirmed to be functionally involved in the generation of ECAD Low/Neg -VIM Pos population, acting upstream of radiation resistance and tumour cell invasion. Conclusion This study highlights the functional significance and complexity of tumour cell remodelling in response to ionising radiation with links to resistance and invasive capacity. An area of less focus in conventional radiotherapy, with the potential to improve treatment outcomes and relapse-free survival.