Epithelial-Mesenchymal Transition during Metastasis of HPV-Negative Pharyngeal Squamous Cell Carcinoma

Abstract EN

In epithelial tumors, a shift towards a mesenchymal phenotype has been associated with increased invasiveness and metastasis.

It is assumed that this phenomenon plays a major role in disease progression and ultimately prognosis.

This study investigated epithelial-mesenchymal transition (EMT) in human papillomavirus- (HPV-) negative pharyngeal squamous cell carcinoma.

Tissue was obtained from one hypopharyngeal primary tumor and a regional lymph node metastasis during surgery with curative intention.

A cell culture was established from the primary tumor and mesenchymal growth conditions were emulated.

Gene expression profiling was performed (Human 8 × 60 K design array, Agilent Technologies) and EMT was assessed by a gene set (MSigDB: M5930, Hallmark_epithelial_mesenchymal_transition), applying gene set expression analysis (GSEA).

Immunohistochemical staining and flow cytometry of CD44 and E-cadherin were compared in primary tumor, metastasis, and cell cultures.

Primary tumor and metastasis were highly positive for CD44.

A loss of E-cadherin occurred in the metastasis.

Flow cytometry showed the appearance of a population without E-cadherin in spheroid colonies.

In GSEA, the EMT phenotype was enriched in the primary tumor compared to metastasis and cell cultures (FDR < 25%, p < 5%).

EMT showed variable expression during metastasis.

It may thereby be a dynamic state in HPV-negative pharyngeal squamous cell carcinoma that is active only during the process of metastasis itself.

Thereby, the primary tumor as well as the metastasis may exhibit fewer EMT properties.