title: miR-200c and phospho-AKT as prognostic factors and mediators of osteosarcoma progression and lung metastasis.
abstract: Lung metastasis is the major cause of death in osteosarcoma patients. However, molecular mechanisms underlying this metastasis remain poorly understood. To identify key molecules related with pulmonary metastasis of pediatric osteosarcomas, we analyzed high-throughput miRNA expression in a cohort of 11 primary tumors and 15 lung metastases. Results were further validated with an independent cohort of 10 primary tumors and 6 metastases. In parallel, we performed immunohistochemical analysis of activated signaling pathways in 36 primary osteosarcomas. Only phospho-AKT associated with lower overall survival in primary tumors, supporting its role in osteosarcoma progression. CTNNB1 expression also associated with lower overall survival but was not strong enough to be considered an independent variable. Interestingly, miR-200c was overexpressed in lung metastases, implicating an inhibitory feed-back loop to PI3K-AKT. Moreover, transfection of miR200c-mimic in U2-OS cells reduced phospho-AKT levels but increased cellular migration and proliferation. Notably, miR-200c expression strongly correlated with miR-141 and with the osteogenic inhibitor miR-375, all implicated in epithelial to mesenchymal transition. These findings contrast epithelial tumors where reduced miR-200c expression promotes metastasis. Indeed, we noted that osteosarcoma cells in the lung also expressed the epithelial marker CDH1, revealing a change in their mesenchymal phenotype. We propose that miR-200c upregulation occurs late in osteosarcoma progression to provide cells with an epithelial phenotype that facilitates their integration in the metastatic lung niche. Thus, our findings identify phospho-AKT in the primary tumor and miR-200c later during tumor progression as prognostic molecules and potential therapeutic targets to prevent progression and metastasis of pediatric osteosarcomas.
authors.affiliation: Pediatric Oncology Unit, Hospital Universitario y Politécnico La Fe, Spain; Clinical and Translational Research in Cancer, Instituto de Investigación Sanitaria La Fe, Spain.
authors.forename: Pablo
authors.initials: P
authors.lastname: Berlanga
authors.affiliation: Laboratory of Cellular and Molecular Biology, Instituto de Investigación Sanitaria La Fe, Spain.
authors.forename: Lisandra
authors.initials: L
authors.lastname: Muñoz
authors.affiliation: Laboratory of Cellular and Molecular Biology, Instituto de Investigación Sanitaria La Fe, Spain.
authors.forename: Marta
authors.initials: M
authors.lastname: Piqueras
authors.affiliation: Laboratory of Cellular and Molecular Biology, Instituto de Investigación Sanitaria La Fe, Spain.
authors.forename: J Antoni
authors.initials: JA
authors.lastname: Sirerol
authors.affiliation: Genomics Unit, Instituto de Investigación Sanitaria La Fe, Spain.
authors.forename: María Dolors
authors.initials: MD
authors.lastname: Sánchez-Izquierdo
authors.affiliation: Biostatistics Unit, Instituto de Investigación Sanitaria La Fe, Spain.
authors.forename: David
authors.initials: D
authors.lastname: Hervás
authors.affiliation: Department of Pathology, Hospital La Fe, Spain.
authors.forename: Miguel
authors.initials: M
authors.lastname: Hernández
authors.affiliation: Department of Pathology, Hospital La Fe, Spain.
authors.forename: Margarita
authors.initials: M
authors.lastname: Llavador
authors.affiliation: Department of Pathology, Instituto Valenciano de Oncología, Spain.
authors.forename: Isidro
authors.initials: I
authors.lastname: Machado
authors.affiliation: Department of Pathology, University of Valencia, Spain.
authors.forename: Antonio
authors.initials: A
authors.lastname: Llombart-Bosch
authors.affiliation: Pediatric Oncology Unit, Hospital Universitario y Politécnico La Fe, Spain; Clinical and Translational Research in Cancer, Instituto de Investigación Sanitaria La Fe, Spain.
authors.forename: Adela
authors.initials: A
authors.lastname: Cañete
authors.affiliation: Pediatric Oncology Unit, Hospital Universitario y Politécnico La Fe, Spain; Clinical and Translational Research in Cancer, Instituto de Investigación Sanitaria La Fe, Spain.
authors.forename: Victoria
authors.initials: V
authors.lastname: Castel
authors.affiliation: Clinical and Translational Research in Cancer, Instituto de Investigación Sanitaria La Fe, Spain; Laboratory of Cellular and Molecular Biology, Instituto de Investigación Sanitaria La Fe, Spain. Electronic address: jaime.fontdemora@gmail.com.
authors.forename: Jaime
authors.initials: J
authors.lastname: Font de Mora
mesh_terms: D000293:Adolescent; D015703:Antigens, CD; D014408:Biomarkers, Tumor / Q000378:metabolism; D015820:Cadherins / Q000378:metabolism; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics; D049109:Cell Proliferation / Q000235:genetics; D002648:Child; D015331:Cohort Studies; D018450:Disease Progression; D004789:Enzyme Activation; D004847:Epithelial Cells / Q000378:metabolism; D005260:Female; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008175:Lung Neoplasms / Q000473:pathology / Q000556:secondary*; D008297:Male; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D012516:Osteosarcoma / Q000235:genetics* / Q000473:pathology*; D010766:Phosphorylation; D011379:Prognosis; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D015203:Reproducibility of Results; D015398:Signal Transduction; D016019:Survival Analysis; D055815:Young Adult
chemical_list: D015703:Antigens, CD; D014408:Biomarkers, Tumor; C513384:CDH1 protein, human; D015820:Cadherins; C533896:MIRN200 microRNA, human; D035683:MicroRNAs; D051057:Proto-Oncogene Proteins c-akt
keywords: Lung metastasis; Mesenchymal to epithelial transition; Pediatric osteosarcoma; Phospho-AKT; miR-200c