Fig. 2: Fra-1 Positively regulates HMGA2 to promote gastric cancer progression.

A Gene Ontology (GO) enrichment analysis of differentially expressed genes identified in the proteomic profiling of gastric cancer cells overexpressing Fra-1. B The top five upregulated genes from the proteomic analysis of gastric cancer cells with Fra-1 overexpression. C–F RT-qPCR experiments were conducted to assess the impact of Fra-1 overexpression and knockdown on the mRNA levels of the top five upregulated proteins: ITSN2, IQCB1, MED18, CYP26B1, and HMGA2 in gastric cancer cells HGC27/AGS. The p-value was calculated through unpaired t-test correction. G, H Western blot analysis was performed to determine the protein expression levels of HMGA2 in gastric cancer cells HGC27/AGS following Fra-1 overexpression or knockdown. I Representative immunohistochemical images showed the expression of HMGA2 in gastric cancer tissues and corresponding paracancerous tissues. 20×: 50 μm; 40×: 20 μm. J Immunohistochemical semi-quantitative statistical chart. Normal: 21 cases; Cancer: 23 cases; The p-value was calculated through unpaired t-test correction. K, L EdU cell proliferation assay combined with flow cytometry was used to evaluate changes in the proliferation ability of gastric cancer cells HGC27/AGS upon Fra-1 overexpression and HMGA2 knockdown. Multiple comparisons were analyzed using ANOVA, and two groups were compared by Wilcoxon rank sum test. M, N A Transwell cell invasion assay was employed to investigate the effects of Fra-1 overexpression and concurrent HMGA2 knockdown on the invasion capacity of gastric cancer cells HGC27/AGS. Scale bar, 100 μm. Multiple comparisons were analyzed using ANOVA, and two groups were compared by Wilcoxon rank sum test. O A scratch healing assay was conducted to assess the impact of Fra-1 overexpression and simultaneous HMGA2 knockdown on the migration ability of the gastric cancer cell line HGC27. Multiple comparisons were analyzed using ANOVA, and two groups were compared by Wilcoxon rank sum test. Scale bar, 200 μm. All experiments were independently repeated three or more times, and the data presented are from representative individual experiments. “ns” indicates no significant difference; “*” indicates p < 0.05; “**“ indicates p < 0.01; “***” indicates p < 0.001; “****“ indicates p < 0.0001.