title: Transcriptomic Profiling Discloses Molecular and Cellular Events Related to Neuronal Differentiation in SH-SY5Y Neuroblastoma Cells.
abstract: Human SH-SY5Y neuroblastoma cells are widely utilized in in vitro studies to dissect out pathogenetic mechanisms of neurodegenerative disorders. These cells are considered as neuronal precursors and differentiate into more mature neuronal phenotypes under selected growth conditions. In this study, in order to decipher the pathways and cellular processes underlying neuroblastoma cell differentiation in vitro, we performed systematic transcriptomic (RNA-seq) and bioinformatic analysis of SH-SY5Y cells differentiated according to a two-step paradigm: retinoic acid treatment followed by enriched neurobasal medium. Categorization of 1989 differentially expressed genes (DEGs) identified in differentiated cells functionally linked them to changes in cell morphology including remodelling of plasma membrane and cytoskeleton, and neuritogenesis. Seventy-three DEGs were assigned to axonal guidance signalling pathway, and the expression of selected gene products such as neurotrophin receptors, the functionally related SLITRK6, and semaphorins, was validated by immunoblotting. Along with these findings, the differentiated cells exhibited an ability to elongate longer axonal process as assessed by the neuronal cytoskeletal markers biochemical characterization and morphometric evaluation. Recognition of molecular events occurring in differentiated SH-SY5Y cells is critical to accurately interpret the cellular responses to specific stimuli in studies on disease pathogenesis.
authors.affiliation: Department of Neuroscience, Biomedicine and Movement, University of Verona, Verona, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Pezzini
authors.affiliation: Department of Biotechnologies, University of Verona, Verona, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Bettinetti
authors.affiliation: Department of Biotechnologies, University of Verona, Verona, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Di Leva
authors.affiliation: Unit for Neuromuscular and Neurodegenerative Disorders, Laboratory of Molecular Medicine, IRCCS Bambino Gesù Children's Hospital, Rome, Italy.
authors.forename: Marzia
authors.initials: M
authors.lastname: Bianchi
authors.affiliation: Applied Research on Cancer-Network (ARC-NET), University of Verona, Verona, Italy.|Aptuit s.r.l., Verona, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Zoratti
authors.affiliation: Unit for Neuromuscular and Neurodegenerative Disorders, Laboratory of Molecular Medicine, IRCCS Bambino Gesù Children's Hospital, Rome, Italy.
authors.forename: Rosalba
authors.initials: R
authors.lastname: Carrozzo
authors.affiliation: Unit for Neuromuscular and Neurodegenerative Disorders, Laboratory of Molecular Medicine, IRCCS Stella Maris, Calambrone-Pisa, Italy.
authors.forename: Filippo M
authors.initials: FM
authors.lastname: Santorelli
authors.affiliation: Department of Biotechnologies, University of Verona, Verona, Italy.
authors.forename: Massimo
authors.initials: M
authors.lastname: Delledonne
authors.affiliation: Medicum, Biochemistry/Developmental Biology Meilahti Clinical Proteomics Core Facility, University of Helsinki, Helsinki, Finland. maciej.lalowski@helsinki.fi.
authors.forename: Maciej
authors.initials: M
authors.lastname: Lalowski
authors.affiliation: Department of Neuroscience, Biomedicine and Movement, University of Verona, Verona, Italy. alessandro.simonati@univr.it.
authors.forename: Alessandro
authors.identifier: http://orcid.org/0000-0001-6908-5418
authors.initials: A
authors.lastname: Simonati
mesh_terms: D002454:Cell Differentiation / Q000187:drug effects* / Q000502:physiology; D045744:Cell Line, Tumor; D020869:Gene Expression Profiling / Q000379:methods; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D009447:Neuroblastoma / Q000188:drug therapy / Q000378:metabolism*; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D014212:Tretinoin / Q000494:pharmacology
chemical_list: D008565:Membrane Proteins; C581991:Slitrk6 protein, human; D014212:Tretinoin
keywords: Axonal elongation; Axonal guidance signalling; Neuronal markers; RNA-seq analysis; SH-SY5Y differentiation; Semaphorins