20.500.12556/RUL-178431 α‑enolase and γ‑enolase expression in enriched S- and N-type SH-SY5Y cells regulatory role of cathepsin X Enolase is well-known for its role in glycolysis but also plays other roles in the central nervous system, including neuronal survival, differentiation, and axonal regeneration. Here, we investigated α- and γ-enolase expression patterns and their association with cathepsin X in distinct SH-SY5Y cell phenotypes. Enriched substrate-adherent S-type cells are characterized by large, flat morphology with extensive cytoplasm and higher expression of vimentin, while neuroblastic N-type are recognized by neurite extensions and higher expression of B-cell lymphoma 2 (Bcl-2) and growth-associated protein-43. We demonstrated that γ-enolase expression was specific to N-type cells, whereas α-enolase expression was not phenotype-specific. Moreover, a shift from ubiquitously expressed α-enolase to neuron-specific γ-enolase was observed during the enrichment and differentiation. Additionally, cathepsin X exhibited higher proteolytic activity in S-type cells. Inhibition of cathepsin X with AMS36 promoted differentiated cell morphology and increased expression of the active form of γ-enolase. Furthermore, AMS36 altered the expression of vimentin and Bcl-2, indicating a regulatory role in neuronal differentiation. Furthermore, AMS36 activated extracellular signal-regulated kinase 1/2 in N-type cells and enhanced the association between γ-enolase and tyrosine receptor kinase in both, suggesting a link between cathepsin X/γ-enolase and the key signaling pathways of differentiation. Our findings underscore the multifaceted role of enolase isoforms in SH-SY5Y cell differentiation, with α-enolase and γ-enolase showing distinct expression patterns in S- and N-type cells. The expression and activity of cathepsin X in S-type cells, along with its regulatory impact on γ-enolase in N-type cells, highlight the importance of these proteins in neuronal differentiation. neuron-like cells neuronal diferentiation SH-SY5Y cell phenotypes enolase isoforms cathepsin X inhibition nevronom podobne celice nevronska diferenciacija fenotipi celic SH-SY5Y izoforme enolaze katepsin X zaviralci katepsin true false true Angleški jezik Slovenski jezik Članek v reviji 2026-01-27 15:56:03 2026-01-27 15:56:07 2026-01-28 04:29:14 0000-00-00 00:00:00 2025 0 0 Str. 10006–10019 iss. 8 Vol. 62 2025 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 577.152.34 1559-1182 10.1007/s12035-025-04898-2 231412483 s12035-025-04898-2_(1).pdf s12035-025-04898-2_(1).pdf 1 7C915C5D65733450A98ED3B27614BD69 a53cfc3d4bf78fdc0f04802682e747456ee37c00cf35edbbd1832b18550d62aa 2c64d388-fb91-11f0-a1ba-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=226313 https://link.springer.com/article/10.1007/s12035-025-04898-2 1 5897cd31-fb90-11f0-a1ba-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=226312 Fakulteta za farmacijo 0 0 0