Objective：To investigate the expression and significance of long non-coding RNA CBR3-AS1（LncRNA CBR3-AS1）in osteosarcoma tissue and cell lines. Methods：The pathological tissue samples and clinical data of 66 patients with osteosarcoma diagnosed and treated in our hospital from January 2012 to January 2014 were retrospectively collected. The expression of LncRNA CBR3-AS1 in pathological tissues was detected by qRT-PCR. The patients were divided into the high expression group and the low expression group. The correlation of the expression of LncRNA CBR3-AS1 and clinical parameters was analyzed by Chi-square test. Kaplan-Meier survival analysis was used to compare the survival rate of osteosarcoma patients with high or low expression of LncRNA CBR3-AS1. U-2 OS cell lines were divided into the LncRNA CBR3-AS1 silent expression group（si-CBR3-AS1 group）and the negative control group（si-NC group）. LncRNA CBR3-AS1 siRNA and si-NC sequences were transfected by LipofectamineTM 3000，respectively. Cell proliferation，cell migration and invasion were measured by MTT and colony formation assay. Apoptotic capacity was measured by cytometry. Expression of carboxyl reductase 3（CBR3）were detected by qRT-PCR and Western blot. Results：LncRNA CBR3-AS1 were highly expressed in osteosarcoma tissues and cell lines（P < 0.001）. LncRNA CBR3-AS1 high expression were significantly correlated with Enneking staging（P < 0.001），distant metastasis（P=0.004）and histological grade（P=0.036）. LncRNA CBR3-AS1 high expression was negatively correlated with overall survival rate in osteosarcoma patients（P < 0.01）， it was an independent prognostic factor for osteosarcoma patients（HR=1.558，95%CI：1.041-2.641，P=0.013）. Cell proliferation and colony formation ability，as well as cell migration and invasion ability in the si-CBR3-AS1 group were significantly lower than those in the si-NC group，and cell apoptosis rate in the si-CBR3-AS1 group was higher than that in the si-NC group. The expression of LncRNA CBR3-AS1 and CBR3 was positively correlated（r=0.44，P=0.036）. The expression of CBR3 protein in the si-CBR3-AS1 group was significantly lower than that in the si-NC group（P < 0.001）. Conclusion：Downregulated expression of LncRNA CBR3-AS1 inhibits cell proliferation，migration and invasion，and promotes apoptosis，which is an independent risk factor affecting the prognosis of osteosarcoma.