Genetic and Molecular Characterization of the Human Osteosarcoma 3AB-OS Cancer Stem Cell Line: A Possible Model for Studying Osteosarcoma Origin and Stemness RICCARDO DI FIORE, 1 DANIELE FANALE, 2 ROSA DRAGO-FERRANTE, 1 FERDINANDO CHIARADONNA, 3 MICHELA GIULIANO, 1 ANNA DE BLASIO, 1 VALERIA AMODEO, 2 LIDIA R. CORSINI, 2 VIVIANA BAZAN, 2 GIOVANNI TESORIERE, 1,4 RENZA VENTO, 1,4 * AND ANTONIO RUSSO 2,4 1 Section of Biochemical Sciences, Department of Experimental Biomedicine and Clinical Neurosciences, Polyclinic, University of Palermo, Palermo, Italy 2 Section of Medical Oncology, Department of Surgical and Oncological Sciences, Polyclinic, University of Palermo, Palermo, Italy 3 Department of Biotechnology and Biosciences, University of Milano-Bicocca, Milan, Italy 4 Institute for Cancer Research and Molecular Medicine and Center of Biotechnology—College of Science and Biotechnology, Temple University, Philadelphia, Pennsylvania Finding new treatments targeting cancer stem cells (CSCs) within a tumor seems to be critical to halt cancer and improve patient survival. Osteosarcoma is an aggressive tumor affecting adolescents, for which there is no second-line chemotherapy. Uncovering new molecular mechanisms underlying the development of osteosarcoma and origin of CSCs is crucial to identify new possible therapeutic strategies. Here, we aimed to characterize genetically and molecularly the human osteosarcoma 3AB-OS CSC line, previously selected from MG63 cells and which proved to have both in vitro and in vivo features of CSCs. Classic cytogenetic studies demonstrated that 3AB-OS cells have hypertriploid karyotype with 71–82 chromosomes. By comparing 3AB-OS CSCs to the parental cells, array CGH, Affymetrix microarray, and TaqMan 1 Human MicroRNA array analyses identified 49 copy number variations (CNV), 3,512 dysregulated genes and 189 differentially expressed miRNAs. Some of the chromosomal abnormalities and mRNA/miRNA expression profiles appeared to be congruent with those reported in human osteosarcomas. Bioinformatic analyses selected 196 genes and 46 anticorrelated miRNAs involved in carcinogenesis and stemness. For the first time, a predictive network is also described for two miRNA family (let-7/98 and miR-29a,b,c) and their anticorrelated mRNAs (MSTN, CCND2, Lin28B, MEST, HMGA2, and GHR), which may represent new biomarkers for osteosarcoma and may pave the way for the identification of new potential therapeutic targets. J. Cell. Physiol. 228: 1189–1201, 2013. ß 2012 Wiley Periodicals, Inc. The authors declare no conflict of interest. Riccardo Di Fiore and Daniele Fanale contributed equally to this work. Additional supporting information may be found in the online version of this article. Contract grant sponsor: Italian Ministry of Education, University and Research (MIUR) ex-60%, 2007. Contract grant sponsor: Innovative Research Projects (University of Palermo, Italy, 2007). Contract grant sponsor: MIUR-PRIN; Contract grant number: 2008P8BLNF (2008). Contract grant sponsor: MIUR; Contract grant number: 867/06/07/2011. Contract grant sponsor: MIUR; Contract grant number: 2223/12/19/2011. Contract grant sponsor: MIUR-PRIN; Contract grant number: 144/01/26/2012. Contract grant sponsor: Italian Ministry of Health (2008 and 2009). *Correspondence to: Renza Vento, Section of Biochemical Sciences, Department of Experimental Biomedicine and Clinical Neurosciences, Polyclinic, University of Palermo, Via del Vespro 129, Palermo 90127, Italy. E-mail: renza.vento@unipa.it Manuscript Received: 4 September 2012 Manuscript Accepted: 18 October 2012 Accepted manuscript online in Wiley Online Library (wileyonlinelibrary.com): 5 November 2012. DOI: 10.1002/jcp.24272 ORIGINAL RESEARCH ARTICLE 1189 Journal of Journal of Cellular Physiology Cellular Physiology ß 2012 WILEY PERIODICALS, INC.