title: Identification and expansion of human osteosarcoma-cancer-stem cells by long-term 3-aminobenzamide treatment.
abstract: A novel cancer stem-like cell line (3AB-OS), expressing a number of pluripotent stem cell markers, was irreversibly selected from human osteosarcoma MG-63 cells by long-term treatment (100 days) with 3-aminobenzamide (3AB). 3AB-OS cells are a heterogeneous and stable cell population composed by three types of fibroblastoid cells, spindle-shaped, polygonal-shaped, and rounded-shaped. With respect to MG-63 cells, 3AB-OS cells are extremely smaller, possess a much greater capacity to form spheres, a stronger self-renewal ability and much higher levels of cell cycle markers which account for G1-S/G2-M phases progression. Differently from MG-63 cells, 3AB-OS cells can be reseeded unlimitedly without losing their proliferative potential. They show an ATP-binding cassette transporter ABCG2-dependent phenotype with high drug efflux capacity, and a strong positivity for CD133, marker for pluripotent stem cells, which are almost unmeasurable in MG-63 cells. 3AB-OS cells are much less committed to osteogenic and adipogenic differentiation than MG-63 cells and highly express genes required for maintaining stem cell state (Oct3/4, hTERT, nucleostemin, Nanog) and for inhibiting apoptosis (HIF-1alpha, FLIP-L, Bcl-2, XIAP, IAPs, and survivin). 3AB-OS may be a novel tumor cell line useful for investigating the mechanisms by which stem cells enrichment may be induced in a tumor cell line. The identification of a subpopulation of cancer stem cells that drives tumorigenesis and chemoresistance in osteosarcoma may lead to prognosis and optimal therapy determination. Expression patterns of stem cell markers, especially CD133 and ABCG2, may indicate the undifferentiated state of osteosarcoma tumors, and may correlate with unfavorable prognosis in the clinical setting.
authors.affiliation: Dipartimento di Scienze Biochimiche, Università degli Studi di Palermo, Policlinico, Palermo, Italy.
authors.forename: Riccardo
authors.initials: R
authors.lastname: Di Fiore
authors.forename: Andrea
authors.initials: A
authors.lastname: Santulli
authors.forename: Rosa Drago
authors.initials: RD
authors.lastname: Ferrante
authors.forename: Michela
authors.initials: M
authors.lastname: Giuliano
authors.forename: Anna
authors.initials: A
authors.lastname: De Blasio
authors.forename: Concetta
authors.initials: C
authors.lastname: Messina
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Pirozzi
authors.forename: Virginia
authors.initials: V
authors.lastname: Tirino
authors.forename: Giovanni
authors.initials: G
authors.lastname: Tesoriere
authors.forename: Renza
authors.initials: R
authors.lastname: Vento
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000378:metabolism; D000293:Adolescent; D000328:Adult; D017209:Apoptosis / Q000502:physiology; D001549:Benzamides / Q000494:pharmacology*; D001562:Benzimidazoles / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D002121:Calcium Channel Blockers / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology; D045744:Cell Line, Tumor / Q000187:drug effects* / Q000502:physiology; D048430:Cell Shape; D002648:Child; D018432:Drug Resistance, Multiple / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005456:Fluorescent Dyes / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D014411:Neoplastic Stem Cells / Q000187:drug effects* / Q000473:pathology / Q000502:physiology*; D012516:Osteosarcoma / Q000378:metabolism* / Q000473:pathology; D020112:Rhodamine 123 / Q000378:metabolism; D014700:Verapamil / Q000378:metabolism; D055815:Young Adult
chemical_list: D018528:ATP-Binding Cassette Transporters; D001549:Benzamides; D001562:Benzimidazoles; D015415:Biomarkers; D002121:Calcium Channel Blockers; D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D020112:Rhodamine 123; C025160:3-aminobenzamide; D014700:Verapamil; C017807:bisbenzimide ethoxide trihydrochloride