Journal: The Journal of Physiological Sciences : JPS
Article Title: EP4 stimulation promotes cell adhesion and migration via IL-6 signaling in oral squamous cell carcinoma
doi: 10.1016/j.jphyss.2026.100057
Figure Lengend Snippet: RNA-seq analysis demonstrates increased IL-6 expression following EP4 stimulation. RNA sequencing was performed to comprehensively analyze gene expression changes in HSC-3 cells treated with ONO-AE1–437 (1 µM) for 3 h. (A) Bar plot of enrichment (Metascape, default) generated from genes meeting the RNA-seq criteria ( p < 0.05 and |log₂FC| ≥ 0.58). Top enriched terms from the differential gene expression list show extracellular matrix (NABA Matrisome associated) at the highest rank, together with cytokine-related and cell motility terms (values plotted as −log10(P)). (B) Enrichment network of EP4-stimulated RNA-seq. Upper panel: Overall enrichment network. Nodes represent enriched terms; edges connect term pairs with similarity > 0.3. Node colors indicate cluster IDs and size reflects hit count. Terms were selected by Metascape’s default sampling. Lower panel：Focused subnetwork of selected clusters：Focused subnetwork of selected clusters. From upper panel, only the terms belonging to the three clusters—NABA Matrisome associated, positive regulation of cell motility, and Cytokine–cytokine receptor interaction—were extracted and redrawn under the same thresholds and styling. Elliptical bands indicate cluster names. (C) A protein-protein interaction (PPI) network was constructed using Metascape. Node size reflects degree (number of edges). This revealed IL-6 as a hub in the PPI network, showing high degree (k = 7) and betweenness (0.286). Enrichment analysis of the network components identified genes involved in the regulation of cell adhesion and migration, which were visualized. The genes marked in blue are associated with the regulation of cell migration, while the genes marked in red are related to the regulation of cell adhesion. MCODE clustering of the PPI network identified three core clusters, one of which included IL-6 as a hub. Colored nodes indicate MCODE-extracted components. (D) Gene Ontology (GO) terms associated with this cluster included several terms related to cell migration and cell adhesion. (E) RNA-sequencing analysis of gene expression changes in HSC-3 cells treated with the EP4 agonist ONO-AE1–437 (1 μM) for 3 h (n = 4). Differentially expressed genes were identified based on normalized read counts obtained from comprehensive transcriptomic profiling. The IL-6 expression data shown were extracted from the RNA-sequencing results and are presented as a representative EP4-responsive gene.
Article Snippet: The concentration of IL-6 in the culture supernatants was measured using a Human IL-6 Quantikine ELISA Kit (R&D Systems, Minneapolis, MN, USA) according to the manufacturer’s protocol .
Techniques: RNA Sequencing, Expressing, Gene Expression, Generated, Sampling, Construct, Migration
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