title: Deregulation of focal adhesion pathway mediated by miR-659-3p is implicated in bone marrow infiltration of stage M neuroblastoma patients.
abstract: To get insights on the metastatic process of human neuroblastoma (NB), the miRNA expression profile of bone marrow (BM)-infiltrating cells has been determined and compared to that of primary tumors.Twenty-two BM-infiltrating cells, 22 primary tumors, and 4 paired samples from patients with metastatic NB aged > 12 months were analyzed for the expression of 670 miRNAs by stem-loop RT-qPCR. The miRNAs whose expression was significantly different were subjected to selection criteria, and 20 selected miRNAs were tested in 10 additional BM-infiltrating cells and primary tumors. Among the miRNAs confirmed to be differentially expressed, miR-659-3p was further analyzed. Transfection of miR-659-3p mimic and inhibitor demonstrated the specific suppression and over-expression, respectively, of the miR-659-3p target gene CNOT1, a regulator of transcription of genes containing AU-rich element (ARE) sequence. Among the ARE-containing genes, miR-659-3p mimic and inhibitor specifically modified the expression of AKT3, BCL2, CYR61 and THSB2, belonging to the focal adhesion pathway. Most importantly, in BM-infiltrating cells CNOT1 expression was significantly higher, and that of AKT3, BCL2, THSB2 and CYR61 was significantly lower than in primary tumors. Thus, our study suggests a role of the focal adhesion pathway, regulated by miR-659-3p through CNOT1, in the human NB metastatic process.
authors.affiliation: U.O.S. Physiopathology of Human Reproduction, IRCCS A.O.U. San Martino-IST, Genova, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Stigliani
authors.affiliation: U.O.S. Physiopathology of Human Reproduction, IRCCS A.O.U. San Martino-IST, Genova, Italy.
authors.forename: Paola
authors.initials: P
authors.lastname: Scaruffi
authors.affiliation: Department of Economy, University of Genoa, Genova, Italy.
authors.forename: Corrado
authors.initials: C
authors.lastname: Lagazio
authors.affiliation: Department of Economy, University of Genoa, Genova, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Persico
authors.affiliation: Laboratory of Oncology, IRCCS Istituto Giannina Gaslini, Genova, Italy.
authors.forename: Barbara
authors.initials: B
authors.lastname: Carlini
authors.affiliation: Laboratory of Molecular Biology, IRCCS Istituto Giannina Gaslini, Genova, Italy.
authors.forename: Luigi
authors.initials: L
authors.lastname: Varesio
authors.affiliation: Laboratory of Oncology, IRCCS Istituto Giannina Gaslini, Genova, Italy.
authors.forename: Fabio
authors.initials: F
authors.lastname: Morandi
authors.affiliation: Laboratory of Molecular Biology, IRCCS Istituto Giannina Gaslini, Genova, Italy.
authors.forename: Martina
authors.initials: M
authors.lastname: Morini
authors.affiliation: Epidemiology, Biostatistics and Committees Unit, IRCCS Istituto Giannina Gaslini, Genova, Italy.
authors.forename: Anna Rita
authors.initials: AR
authors.lastname: Gigliotti
authors.affiliation: Neuroblastoma Laboratory, Pediatric Research Institute, Fondazione Città della Speranza, Padova, Italy.
authors.forename: Maria Rosaria
authors.initials: MR
authors.lastname: Esposito
authors.affiliation: Pediatric Hematology and Oncology Division, Padova University Hospital, Padova, Italy.
authors.forename: Elisabetta
authors.initials: E
authors.lastname: Viscardi
authors.affiliation: UOS Divisione Oncoematologia Pediatrica, Ospedale Civile di Pescara, Pescara, Italy.
authors.forename: Valerio
authors.initials: V
authors.lastname: Cecinati
authors.affiliation: Oncology Unit, IRCCS Istituto Giannina Gaslini, Genova, Italy.
authors.forename: Massimo
authors.initials: M
authors.lastname: Conte
authors.affiliation: Laboratory of Oncology, IRCCS Istituto Giannina Gaslini, Genova, Italy.
authors.forename: Maria Valeria
authors.initials: MV
authors.lastname: Corrias
mesh_terms: D019046:Bone Marrow Neoplasms / Q000235:genetics* / Q000556:secondary; D045744:Cell Line, Tumor; D002648:Child; D002675:Child, Preschool; D005260:Female; D022001:Focal Adhesions / Q000502:physiology; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000502:physiology*; D006801:Humans; D007223:Infant; D008297:Male; D035683:MicroRNAs / Q000235:genetics*; D009447:Neuroblastoma / Q000235:genetics* / Q000473:pathology; D060888:Real-Time Polymerase Chain Reaction; D014157:Transcription Factors / Q000378:metabolism; D014162:Transfection
chemical_list: C528690:CNOT1 protein, human; C533992:MIRN659 microRNA, human; D035683:MicroRNAs; D014157:Transcription Factors
keywords: bone marrow; focal adhesion; metastases; miRNA; neuroblastoma