EFNA2 mediates stiffness-regulated hypopharyngeal cancer progression.

Abstract

Objective: Hypopharyngeal cancer (HPC) originates from the hypopharyngeal mucosa with poor prognosis. The stiffness of extracellular matrix (ECM) is closely associated with tumor progression and prognosis. This study aimed to investigate the impact of matrix stiffness and identify molecular markers that are relevant to the prognosis of HPC, with the goal of improving prognosis.

Methods: Immunohistochemical and cervical enhanced CT data were used to analyze the correlation between CT value, matrix stiffness, and prognosis, and to establish prognosis prediction model of HPC. Cell culture models with different matrix stiffness were constructed by polypropylene hydrogel. Western blotting, clone formation, EDU, and Transwell were used to investigate the effects of matrix stiffness on proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT). High-throughput sequencing was used to identify differential genes in HPC cells cultured in different matrix stiffness. Gene editing technique, in vivo subcutaneous tumorigenesis studies, and western blotting were conducted to investigate molecular mechanisms underlying the impact of high-stiffness matrix on HPC, and the influence of EFNA2 on proliferation, migration, and EMT.

Results: The arterial CT value was positively correlated with matrix stiffness. Matrix stiffness is associated with lymph node metastasis, therapeutic effect, and prognosis. Additionally, metastatic lymph nodes in HPC patients had higher CT values compared to those in patients with nasopharyngeal carcinoma (NPC). High-stiffness ECM could promote the proliferation, migration and EMT of HPC cells. Mechanistically, highstiffness ECM infuenced EFNA2 expression to promote proliferation, migration, EMT, and tumor growth in vivo.

Conclusion: EFNA2 and high matrix stiffness jointly promoted the malignant phenotype in HPC. EFNA2 may serve as potential therapeutic target for high matrix stiffnessinduced HPC progression.