Cultivation of Head and Neck Squamous Cell Carcinoma Cells with Wound Fluid Leads to Cisplatin Resistance via Epithelial-Mesenchymal Transition Induction.

Till Jasper Meyer,Marietta Herrmann,Pascal Ickrath,Agmal Scherzad,Rudolf Hagen,Helena Moratin,Manuel Stöth,Stephan Hackenberg,Norbert Kleinsasser

doi:10.3390/ijms22094474

Abstract

Locoregional recurrence is a major reason for therapy failure after surgical resection of head and neck squamous cell carcinoma (HNSCC). The physiological process of postoperative wound healing could potentially support the proliferation of remaining tumor cells. The aim of this study was to evaluate the influence of wound fluid (WF) on the cell cycle distribution and a potential induction of epithelial-mesenchymal transition (EMT). To verify this hypothesis, we incubated FaDu and HLaC78 cells with postoperative WF from patients after neck dissection. Cell viability in dependence of WF concentration and cisplatin was measured by flow cytometry. Cell cycle analysis was performed by flow cytometry and EMT-marker expression by rtPCR. WF showed high concentrations of interleukin (IL)-6, IL-8, IL-10, CCL2, MCP-1, EGF, angiogenin, and leptin. The cultivation of tumor cells with WF resulted in a significant increase in cell proliferation without affecting the cell cycle. In addition, there was a significant enhancement of the mesenchymal markers Snail 2 and vimentin, while the expression of the epithelial marker E-cadherin was significantly decreased. After cisplatin treatment, tumor cells incubated with WF showed a significantly higher resistance compared with the control group. The effect of cisplatin-resistance was dependent on the WF concentration. In summary, proinflammatory cytokines are predominantly found in WF. Furthermore, the results suggest that EMT can be induced by WF, which could be a possible mechanism for cisplatin resistance.

Highlights

Every year more than 800,000 people worldwide develop head and neck squamous cell carcinoma (HNSCC) and more than half of these patients die from this disease [1]
Already during surgical tumor resection, the wound healing process starts. This is orchestrated by the activation of various immune cells, such as neutrophils, macrophages, and natural killer cells, and of stromal cells, such as fibroblasts or mesenchymal stem cells and secretion of different cytokines and growth factors [3,4]
We could show that wound fluid (WF) significantly increased the expression of Snail of E-ca cultivation withlines

Summary

Introduction

Every year more than 800,000 people worldwide develop head and neck squamous cell carcinoma (HNSCC) and more than half of these patients die from this disease [1].Risk factors include nicotine and alcohol consumption as well as infection with human papilloma virus [2]. Already during surgical tumor resection, the wound healing process starts. This is orchestrated by the activation of various immune cells, such as neutrophils, macrophages, and natural killer cells, and of stromal cells, such as fibroblasts or mesenchymal stem cells and secretion of different cytokines and growth factors [3,4]. Many of these components are found in surgical wound fluid (WF). One component of phenotypic change during wound healing is the epithelial-mesenchymal transition (EMT). The cells gain the ability to detach from their physiological cell complex, break through the basement membrane, and penetrate into the surrounding tissue as well as into lymphatic and blood vessels, where they adapt to new conditions [6,7]

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