title: Prognostic significance of miR-34a in Ewing sarcoma is associated with cyclin D1 and ki-67 expression.
abstract: BACKGROUND
  At diagnosis, identification of reliable biological indicators of prognosis to allow stratification of patients according to different risks is an important but still unresolved aspect in the treatment of Ewing sarcoma (EWS) patients. This study aimed to explore the role of miR-34A expression on prognosis of EWS patients.
  
  
  METHODS
  Specimens from 109 patients with non-metastatic EWS treated at the Rizzoli Institute with neoadjuvant chemotherapy (protocols ISG/SSGIII, EW-1, EW-2, EW-REN2, EW-REN3, EW-PILOT) and 17 metastases were studied. Sixty-eight patients (62%) remained disease-free and 41 (38%) relapsed (median follow-up: 67 months, range 9-241 months). Expression of miR-34a and of some of its targets (cyclin D1, bcl-2, SIRT1 and YY1) was evaluated by qRT-PCR using TaqMan MicroRNA Assays and/or by immunohistochemistry on tissue microarrays from the same patients.
  
  
  RESULTS
  High expression of miR-34a in localized tumors was significantly related to better event-free and overall survival (P = 0.004). Relevance of miR-34a was confirmed by using different calibrators (normal mesenchymal stem cells and different normal tissues). By multivariate Cox regression analysis, low miR-34a expression as well as nontotal necrosis and high levels of lactate dehydrogenase were all confirmed as independent risk factors associated with poor outcome. Expression of miR-34a was lower in metastases than in primary tumors. It inversely correlated with expression of cyclin D1 and Ki-67.
  
  
  CONCLUSIONS
  By demonstrating its relationship with clinical outcome, we propose evaluation of miR-34a at diagnosis of EWS patients to allow early risk stratification. Validation of these results would nonetheless ultimately need a prospective assessment.
authors.affiliation: Experimental Oncology Lab, CRS Development of Biomolecular Therapies, Rizzoli Institute, Bologna.
authors.forename: M T
authors.initials: MT
authors.lastname: Marino
authors.affiliation: Experimental Oncology Lab, CRS Development of Biomolecular Therapies, Rizzoli Institute, Bologna.
authors.forename: A
authors.initials: A
authors.lastname: Grilli
authors.affiliation: Experimental Oncology Lab, CRS Development of Biomolecular Therapies, Rizzoli Institute, Bologna.
authors.forename: C
authors.initials: C
authors.lastname: Baricordi
authors.affiliation: Experimental Oncology Lab, CRS Development of Biomolecular Therapies, Rizzoli Institute, Bologna.
authors.forename: M C
authors.initials: MC
authors.lastname: Manara
authors.affiliation: Experimental Oncology Lab, CRS Development of Biomolecular Therapies, Rizzoli Institute, Bologna.
authors.forename: S
authors.initials: S
authors.lastname: Ventura
authors.affiliation: Experimental Oncology Lab, CRS Development of Biomolecular Therapies, Rizzoli Institute, Bologna.
authors.forename: R S
authors.initials: RS
authors.lastname: Pinca
authors.affiliation: Department of Hematology, Oncology and Molecular Medicine, Istituto Superiore Sanità, Rome.
authors.forename: M
authors.initials: M
authors.lastname: Bellenghi
authors.affiliation: Hematology Unit with Bone Marrow Transplantation, University of Palermo, Palermo.
authors.forename: M
authors.initials: M
authors.lastname: Calvaruso
authors.affiliation: Department of Hematology, Oncology and Molecular Medicine, Istituto Superiore Sanità, Rome.
authors.forename: G
authors.initials: G
authors.lastname: Mattia
authors.affiliation: III Orthopaedic Clinic, Rizzoli Institute, Bologna.
authors.forename: D
authors.initials: D
authors.lastname: Donati
authors.affiliation: Hematology Unit with Bone Marrow Transplantation, University of Palermo, Palermo.
authors.forename: C
authors.initials: C
authors.lastname: Tripodo
authors.affiliation: Experimental Oncology Lab, CRS Development of Biomolecular Therapies, Rizzoli Institute, Bologna.
authors.forename: P
authors.initials: P
authors.lastname: Picci
authors.affiliation: Chemotherapy Section, Rizzoli Institute, Bologna, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Ferrari
authors.affiliation: Experimental Oncology Lab, CRS Development of Biomolecular Therapies, Rizzoli Institute, Bologna. Electronic address: katia.scotlandi@ior.it.
authors.forename: K
authors.initials: K
authors.lastname: Scotlandi
mesh_terms: D000328:Adult; D000369:Aged, 80 and over; D019938:Cyclin D1 / Q000096:biosynthesis*; D018572:Disease-Free Survival; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D006836:Hydro-Lyases / Q000096:biosynthesis; D019394:Ki-67 Antigen / Q000096:biosynthesis*; D008297:Male; D035683:MicroRNAs / Q000096:biosynthesis* / Q000235:genetics; D008875:Middle Aged; D020360:Neoadjuvant Therapy; D009362:Neoplasm Metastasis; D011379:Prognosis; D012512:Sarcoma, Ewing / Q000235:genetics* / Q000473:pathology / Q000628:therapy*; D016896:Treatment Outcome
chemical_list: D019394:Ki-67 Antigen; C522425:MIRN34 microRNA, human; D035683:MicroRNAs; D019938:Cyclin D1; D006836:Hydro-Lyases; C045581:lactate dehydratase
keywords: Ewing sarcoma; Ki-67; cyclin D1; miR-34a; prognostic biomarkers