A High-Content Screening Approach to Identify MicroRNAs Against Head and Neck Cancer Cell Survival and EMT in an...

Area of Science:

• Oncology
• Molecular Biology
• Cell Biology

Background:

• Head and neck squamous cell carcinoma (HNSCC) is a common cancer where metastasis, driven by inflammation-induced epithelial-mesenchymal transition (EMT), is a primary cause of mortality.
• Inflammation promotes EMT via NF-κB-mediated Snail stabilization, highlighting a critical mechanism in HNSCC progression.

Purpose of the Study:

• To investigate the role of microRNAs (miRs) in regulating cell survival and EMT in HNSCC cells within an inflammatory microenvironment.
• To identify specific miRs and signaling pathways that can counteract inflammation-driven HNSCC progression.

Main Methods:

• High-content screening (HCS) was used to assess morphometric and molecular changes (Snail/Slug) in TNF-α-stimulated HNSCC cells.
• 31 distinct miRs were screened for their effects on cell survival and EMT, followed by bioinformatic analysis of enriched signaling pathways.
• Gene silencing of key pathway components (RELA, AKT1, CTNNB1) was performed to validate miR-targeted pathways.

Main Results:

• TNF-α (20 ng/ml) effectively induced EMT markers, including morphological changes, Snail/Slug expression, and increased cell migration.
• miRs with anti-survival and anti-EMT effects targeted pathways including NF-κB, PI3K/AKT, and Wnt/beta-catenin.
• Silencing RELA, AKT1, or CTNNB1 significantly reduced HNSCC cell survival and/or Snail/Slug expression under inflammatory conditions.

Conclusions:

• The HCS approach successfully identified miRs that inhibit HNSCC cell survival and EMT in an inflammatory context.
• Key signaling pathways (NF-κB, PI3K/AKT, Wnt/beta-catenin) and their molecular targets are crucial mediators of inflammation-induced HNSCC progression.
• These findings provide a foundation for developing targeted therapies against HNSCC by modulating these identified pathways and miRs.