Abstract
Hyaluronic acid (HA), a major glycosaminoglycan in the extracellular matrix, is synthesized by hyaluronan synthases (HAS1, HAS2, and HAS3). Dysregulated HA synthesis promotes tumor progression by influencing signaling pathways involving HA receptors such as CD44, HMMR, STAB2, LAYN, and TLR4. This study aimed to investigate the effects of Urtica dioica agglutinin (UDA) on the expression of HA-synthesizing and key HA receptor genes in PC3, BT-549, and HUVEC cell lines. Results showed that UDA significantly downregulated HAS2 (p ≤ 0.001) and HAS3 (p ≤ 0.001) expression in PC3 and BT-549 cell lines, along with HMMR (p ≤ 0.01), STAB2 (p ≤ 0.05), LAYN (p ≤ 0.05), and TLR4 (p ≤ 0.001), while CD44 expression was upregulated (p ≤ 0.001). In HUVEC cells, UDA showed no significant alteration in HAS2 or HAS3 expression (p > 0.05); however, it significantly reduced TLR4 expression (p ≤ 0.05). Molecular docking analysis revealed strong binding affinities between UDA and TLR4 (Docking-score: -342.79 kcal/mol, Ligand-RMSD: 30.56Å), HAS2 (Docking-score: -320.84 kcal/mol, Ligand-RMSD: 50.97Å), and HAS3 (Docking-score: -314.96 kcal/mol, Ligand-RMSD: 58.44Å), supporting its potential interaction with HA-associated proteins. Hence, in HUVEC cells, UDA's effect on TLR4 indicates a direct interaction rather than an indirect effect through HA-related pathways. These findings suggest that UDA may modulate HA-synthesis and HA-receptor genes expression in cancer cells. It provides insights into the molecular mechanisms underlying UDA's anti-cancer properties and its potential as a therapeutic agent targeting HA signaling in cancer. Further research is needed to elucidate these mechanisms and explore UDA's clinical applications.