MicroRNA-29b-1 impairs in vitro cell proliferation, self‑renewal and chemoresistance of human osteosarcoma 3AB-OS cancer stem cells

Di Fiore,Riccardo; Drago-Ferrante,Rosa; Pentimalli,Francesca; Di Marzo,Domenico; Forte,Iris  Maria; D'Anneo,Antonella; Carlisi,Daniela; De Blasio,Anna; Giuliano,Michela; Tesoriere,Giovanni; Giordano,Antonio; Vento,Renza

doi:10.3892/ijo.2014.2618

MicroRNA-29b-1 impairs in vitro cell proliferation, self‑renewal and chemoresistance of human osteosarcoma 3AB-OS cancer stem cells

Authors:
- Riccardo Di Fiore
- Rosa Drago-Ferrante
- Francesca Pentimalli
- Domenico Di Marzo
- Iris Maria Forte
- Antonella D'Anneo
- Daniela Carlisi
- Anna De Blasio
- Michela Giuliano
- Giovanni Tesoriere
- Antonio Giordano
- Renza Vento
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Affiliations: Laboratory of Biochemistry, Department of Biological, Chemical and Pharmaceutical Sciences and Technologies, University of Palermo, Polyclinic, Palermo, Italy, INT-CROM, ‘Pascale Foundation’, National Cancer Institute - Cancer Research Center, Mercogliano, Avellino, Italy, Laboratory of Biochemistry, Department of Experimental Biomedicine and Clinical Neurosciences, University of Palermo, Polyclinic, Palermo, Italy, Institute for Cancer Research and Molecular Medicine and Center of Biotechnology - College of Science and Biotechnology, Temple University, Philadelphia, PA, USA
Published online on: August 22, 2014 https://doi.org/10.3892/ijo.2014.2618
Pages: 2013-2023

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Abstract

Osteosarcoma (OS) is the most common type of bone cancer, with a peak incidence in the early childhood. Emerging evidence suggests that treatments targeting cancer stem cells (CSCs) within a tumor can halt cancer and improve patient survival. MicroRNAs (miRNAs) have been implicated in the maintenance of the CSC phenotype, thus, identification of CSC-related miRNAs would provide information for a better understanding of CSCs. Downregulation of miRNA-29 family members (miR-29a/b/c; miR‑29s) was observed in human OS, however, little is known about the functions of miR-29s in human OS CSCs. Previously, during the characterization of 3AB-OS cells, a CSC line selected from human OS MG63 cells, we showed a potent downregulation of miR-29b. In this study, after stable transfection of 3AB-OS cells with miR-29b-1, we investigated the role of miR-29b-1 in regulating cell proliferation, sarcosphere-forming ability, clonogenic growth, chemosensitivity, migration and invasive ability of 3AB-OS cells, in vitro. We found that, miR-29b-1 overexpression consistently reduced both, 3AB-OS CSCs growth in two- and three-dimensional culture systems and their sarcosphere- and colony-forming ability. In addition, while miR-29b-1 overexpression sensitized 3AB-OS cells to chemotherapeutic drug-induced apoptosis, it did not influence their migratory and invasive capacities, thus suggesting a context-depending role of miR-29b-1. Using publicly available databases, we proceeded to identify potential miR-29b target genes, known to play a role in the above reported functions. Among these targets we analyzed CD133, N-Myc, CCND2, E2F1 and E2F2, Bcl-2 and IAP-2. We also analyzed the most important stemness markers as Oct3/4, Sox2 and Nanog. Real-time RT-PCR and western-blot analyses showed that miR-29b-1 negatively regulated the expression of these markers. Overall, the results show that miR-29b-1 suppresses stemness properties of 3AB-OS CSCs and suggest that developing miR-29b-1 as a novel therapeutic agent might offer benefits for OS treatment.

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