Deregulation of focal adhesion pathway mediated by miR-659-3p is implicated in bone marrow infiltration of stage M neuroblastoma patients
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Sara Stigliani1,*, Paola Scaruffi1,*, Corrado Lagazio2, Luca Persico2, Barbara Carlini3, Luigi Varesio4, Fabio Morandi3, Martina Morini4, Anna Rita Gigliotti5, Maria Rosaria Esposito6, Elisabetta Viscardi7, Valerio Cecinati8, Massimo Conte9, Maria Valeria Corrias3
1U.O.S. Physiopathology of Human Reproduction, IRCCS A.O.U. San Martino-IST, Genova, Italy
2Department of Economy, University of Genoa, Genova, Italy
3Laboratory of Oncology, IRCCS Istituto Giannina Gaslini, Genova, Italy
4Laboratory of Molecular Biology, IRCCS Istituto Giannina Gaslini, Genova, Italy
5Epidemiology, Biostatistics and Committees Unit, IRCCS Istituto Giannina Gaslini, Genova, Italy
6Neuroblastoma Laboratory, Pediatric Research Institute, Fondazione Città della Speranza, Padova, Italy
7Pediatric Hematology and Oncology Division, Padova University Hospital, Padova, Italy
8UOS Divisione Oncoematologia Pediatrica, Ospedale Civile di Pescara, Pescara, Italy
9Oncology Unit, IRCCS Istituto Giannina Gaslini, Genova, Italy
*These authors have contributed equally to this work
Maria Valeria Corrias, e-mail: firstname.lastname@example.org
Keywords: neuroblastoma, focal adhesion, metastases, bone marrow, miRNA
Received: February 09, 2015 Accepted: April 08, 2015 Published: April 20, 2015
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.
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