Numerous pathways were enriched by down-regulated genes; the most significant ones were the insulin, Wnt and integrins signaling pathways, MAPK, cytokine-cytokine receptor interaction, tight junction and focal adhesion, all representing proteins that are highly expressed in malignancies.
Among these, metabolic pathways were the most enriched, with 1024 associated genes, followed by MAPK signaling, actin cytoskeleton regulation, secondary metabolite biosynthesis, focal adhesion, insulin signaling, calcium signaling, cytokine-cytokine receptor interaction, tight junctions, phagosomes, and adipocytokine signaling pathways (Table 3).
IL-8, COL1A1, COL1A2, THBS2, SPP1, COL6A3 and SFRP4 were mapped to several highly impacted pathways: leukocyte transendothelial migration, extracellular matrix receptor interaction, tight junction, epithelial cell signaling in H. pylori infection, TGFβ signaling pathway, toll-like receptor signaling and Wnt signaling (Table 5).
GSEA showed enrichment of genes involved in fatty acid and amino acid metabolism and oxidative phosphorylation similar to the genes down-regulated at 6 h, but also showed decreases in drug and xenobiotic metabolism, extracellular matrix interaction, tight junction formation, and hedgehog and TGFβ signaling in agreement with the progressive epithelial-to-mesenchymal transformation.
Sixteen pathways were selected including mTOR signaling, insulin-like growth factor, MAPK signaling and TGF- β signaling and other pathways such as ubiquitin mediated proteolysis, focal adhesion, Wnt signaling pathway, regulation of actin cytoskeleton, cytokine-cytokine receptor interaction, tight junction, and cell cycle which are known to be involved in cell and tissue structure.
This makes the inter-filament interactions tight, and turns the egg envelope into a hard and tough structure.
These pathways include metabolic, ECM-receptor interactions, tight junctions, biosynthesis of unsaturated fatty acids, ubiquitin mediated proteolysis pathways etc.
The LFA-1/ICAM interaction promotes tight adhesion between activated leukocytes and the endothelium, as well as between T cells and antigen-presenting cells.
This may point toward the interaction between tight junction assembly and epithelial apical basal polarization.
Interestingly, the Mw reported by SEC-MALS remained constant over a wide NEMO355 concentration range, demonstrating that the interaction is tight in accordance with previous studies [ 20, 29].
The protein protein interaction in tight junction proteins can link to the actin cytoskeleton, serving as a maintainer of cell structure (Yilmaz and Christofori, 2009).
