Phosphoinositide-specific phospholipase C isoforms are conveyed by osteosarcoma-derived extracellular vesicles
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RESEARCH ARTICLE
Phosphoinositide-specific phospholipase C isoforms are conveyed by osteosarcoma-derived extracellular vesicles Enrica Urciuoli 1 & Martina Leopizzi 2 & Valeria Di Maio 2 & Stefania Petrini 3 & Valentina D’Oria 3 & Ezio Giorda 4 & Marco Scarsella 4 & Carlo Della Rocca 2 & Vincenza Rita Lo Vasco 5 & Barbara Peruzzi 1 Received: 26 May 2020 / Accepted: 14 June 2020 # The International CCN Society 2020
Abstract Cancer cells are able to release high amounts of extracellular vesicles, thereby conditioning the normal cells in the surrounding tissue and/or in distant target organs. In the context of bone cancers, previous studies suggested that osteosarcoma cancer cells produce transforming extracellular vesicles able to induce a tumour-like phenotype in normal recipient cells. Indeed, phosphoinositide-specific phospholipase C (PI-PLC) enzymes are differentially expressed in osteosarcoma cell lines with increasing aggressiveness, thus providing helpful insights to better define their role and functions in this bone tumour. By confocal microscopy analysis, we demonstrated that osteosarcoma-derived extracellular vesicles convey all the assessed PI-PLC isoforms, and that they localize into cell membrane bubble-like structures, resembling extracellular vesicles about to be released, as conveyed and/or membrane protein. Cytofluorimetric analysis confirmed the presence of PI-PLC isoforms in the extracellular vesicles collected from conditioned media of osteosarcoma cells. These findings suggest the feasibility to use circulating extracellular vesicles as biomarkers of osteosarcoma progression and/or the monitoring of this distressing disease. Keywords Osteosarcoma . PI-PLC . Extracellular vesicles . Confocal microscope . FACS analysis
Introduction The phosphoinositide-specific phospholipase C (PI-PLC) mammal family includes a related group of complex and multi-domain enzymes that cover a various spectrum of regulatory interactions, including direct binding to G protein Enrica Urciuoli and Martina Leopizzi contributed equally to this work. * Barbara Peruzzi [email protected] 1
Multifactorial Disease and Complex Phenotype Research Area, Research Centre, Bambino Gesù Children’s Hospital, Rome, Italy
2
Department of Medico-Surgical Sciences and Biotechnology, Polo Pontino, Sapienza University, Latina, Rome, Italy
3
Confocal Microscopy Core Facility, Research Centre, Bambino Gesù Children’s Hospital, Rome, Italy
4
Flow Cytometry Core Facility, Research Centre, Bambino Gesù Children’s Hospital, Rome, Italy
5
Department of Biomedical, Metabolic and Neural Sciences, Section of Human Morphology, University of Modena and Reggio Emilia, Modena, Italy
subunits, small GTPases from Rho and Ras families, receptor and non-receptor tyrosine kinases, and lipid components of cellular membranes (Rhee et al. 1991). PLC enzymes are thirteen isoforms classified on the basis of domain structure and mechanism of recruitment into six subfamilies: β (1–4), γ (1– 2), δ (1,3,4), ε, ζ, and η (1–2) (Suh et al. 2008). PLC en
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