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. 2024 Feb 6;2024:4123737. doi: 10.1155/2024/4123737

Figure 4.

Figure 4

Fibroblast-derived FERMT2 promotes epithelial mesenchymal transition (EMT) in GC cells in vitro. (a) Protein-protein interaction (PPI) network for FERMT2 was constructed in GeneMANIA. Different colors of the network edge indicate the bioinformatics methods applied, including physical interaction, coexpression, and predicted. The link between FERMT2 and ZEB1 was highlighted with a red box. (b) Schematic diagram of noncontact coculture of fibroblasts and GC cells. (c, d) Western blotting results comparing FERMT2 and β-actin bands before and after si-FERMT2 (CAFs) (paired t-test). Asterisk represents P value (∗∗∗∗P < 0.0001). (e) The clone formation capacity of GC cells under different interventions. (f) Schematic diagram of subcutaneous tumor models. (g) Xenograft mouse tumors (n = 6 mice per group). (h, i) Volumes of xenograft tumors measured twice a week and weights of xenograft tumors at completion of the study. (j) Gene set enrichment analysis (GSEA) for FERMT2. The enriched gene sets in hallmark collection by the high FERMT2 expression samples. The (k, l) wound healing cell migration and (m, n) Transwell migration assays indicate that downregulation of FERMT2 in CAFs weakens the migratory and invasive capacities of GC cells (ANOVA) (scale bars = 100 μm). (o) Correlation of FERMT2 expression with EMT markers in TCGA-STAD. Spearman method was used. (p, q) The expression of the EMT markers with GC cells was examined by Western blotting after the treatment of the fibroblasts with NC and si-FERMT2 constructs (ANOVA) (scale bars = 50 μm). (r, s) Interfering with FERMT2 expression in vivo inhibits EMT (scale bars = 20 μm). One-way ANOVA was conducted. Asterisk represents P value (∗P < 0.05). (t) mIHC staining images of GC section (merge and FERMT2-2×, E-cadherin and N-cadherin -20x) (scale bars = 50 μm and 500 μm). Each experiment was independently performed in triplicate. Each error bar indicates the variation between the means of three independent experiments. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, and ∗∗∗∗P < 0.0001.