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Centromere protein F (CENPF) plays a key role in the regulation of the cell cycle. The present study revealed that CENPF was overexpressed in a variety of tumors and associated with the poor prognosis of osteosarcoma. The mRNA expression levels of CENPF were analyzed using the Gene Expression Profiling Interactive Analysis database and the protein levels of CENPF were detected in the specimens from patients with osteosarcoma using immunohistochemistry. Cell proliferation, cell cycle and flow cytometry assays were performed after the transfection of control or CENPF plasmids into osteosarcoma cells. A xenografts assay was used to determine the effects of CENPF on tumor growth in vivo . The results showed that CENPF was upregulated in osteosarcoma tissues and associated with high-grade tumor stage (P=0.023) and intraglandular dissemination (P=0.046). The transfection-induced depletion of CENPF in human osteosarcoma MG-63 and U-2 OS cell lines inhibited cell proliferation, stimulated apoptosis and induced cell cycle arrest. Induced CENPF depletion in MG-63 cells inhibited tumor growth of osteosarcoma cells in mice. These findings suggested that elevated CENPF levels contributed to increased cell proliferation by mediating apoptosis and cell cycle in osteosarcoma. Therefore, CENPF might be a potential biomarker for poor prognosis of osteosarcoma., Competing Interests: The authors declare that they have no competing interests., (Copyright: © Zou et al.)