title: Proteomics and phosphoproteomics provide insights into the mechanism of action of a novel pyrazolo[3,4-d]pyrimidine Src inhibitor in human osteosarcoma.
abstract: Osteosarcoma (OS) is a highly malignant bone tumour, affecting mainly children and young adults between 10 and 20 years of age. It represents the most frequent primitive malignant tumour of the skeletal system and is characterized by an extremely aggressive clinical course, with rapid development of lung metastases. In the last few years, targeting Src in the treatment of OS has become one of the major challenges in the development of new drugs, since an elevated Src kinase activity has been associated with the development and the maintenance of the OS malignant phenotype. Recently, SI-83, a novel pyrazolo[3,4-d]pyrimidine derivate Src inhibitor, was selected as a promising OS therapeutic drug because of its elevated anti-tumour effects toward human OS. In the present study, gel-based proteomics and phosphoproteomics revealed significant changes in proteins involved in many cancer related processes. We got insight into SI-83 proapoptotic and antiproliferative properties (overrepresentation of GRIA1, GRP78, and CALR and underrepresentation of NPM1, RCN, and P4HB). Nevertheless, the most significant findings of our work are the SI-83 induced dephosphorylation of ARPC5L, a subunit of the actin related Arp2/3 complex, and the decrease of other cytoskeleton proteins. These data, together with a dramatic impairment of SaOS-2 cell migration and adhesion, suggest that SI-83 may have antimetastatic features that enhance its use as a potent OS chemotherapeutic drug.
authors.affiliation: Dipartimento di Biotecnologie, Chimica e Farmacia, Università degli Studi di Siena, via Fiorentina 1, 53100 Siena, Italy. annalisa.santucci@unisi.it.
authors.forename: Giulia
authors.initials: G
authors.lastname: Bernardini
authors.forename: Marcella
authors.initials: M
authors.lastname: Laschi
authors.forename: Tommaso
authors.initials: T
authors.lastname: Serchi
authors.forename: Adriano
authors.initials: A
authors.lastname: Spreafico
authors.forename: Maurizio
authors.initials: M
authors.lastname: Botta
authors.forename: Silvia
authors.initials: S
authors.lastname: Schenone
authors.forename: Simona
authors.initials: S
authors.lastname: Arena
authors.forename: Michela
authors.initials: M
authors.lastname: Geminiani
authors.forename: Andrea
authors.initials: A
authors.lastname: Scaloni
authors.forename: Giulia
authors.initials: G
authors.lastname: Collodel
authors.forename: Maurizio
authors.initials: M
authors.lastname: Orlandini
authors.forename: Neri
authors.initials: N
authors.lastname: Niccolai
authors.forename: Annalisa
authors.initials: A
authors.lastname: Santucci
mesh_terms: D051376:Actin-Related Protein 2-3 Complex / Q000378:metabolism; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D000091342:Endoplasmic Reticulum Chaperone BiP; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D006801:Humans; D000090243:Nucleophosmin; D012516:Osteosarcoma / Q000378:metabolism* / Q000473:pathology; D010766:Phosphorylation; D040901:Proteomics; D011720:Pyrazoles / Q000494:pharmacology*; D011743:Pyrimidines / Q000494:pharmacology*
chemical_list: C573916:ARPC5 protein, human; D051376:Actin-Related Protein 2-3 Complex; D000970:Antineoplastic Agents; D000091342:Endoplasmic Reticulum Chaperone BiP; C000717247:HSPA5 protein, human; C000716967:NPM1 protein, human; D011720:Pyrazoles; D011743:Pyrimidines; C529160:SI-83 compound; D000090243:Nucleophosmin