In our results, we found lncRNA CBR3-AS1 was highly-expressed in osteosarcoma tissues and cell lines, and associated with Enneking stage, distant metastasis and histological grade. Survival analysis indicated that the high-expression of lncRNA CBR3-AS1 was an independent poor prognostic factor for osteosarcoma patients. Loss-of-function studies showed knockdown of lncRNA CBR3-AS1 suppressed osteosarcoma cells proliferation, migration and invasion, and promotes cells apoptosis, but had no effect on cell-cycle distribution. There was no association between lncRNA CBR3-AS1 and CBR3 expression in osteosarcoma tissues, and knockdown of lncRNA CBR3-AS1 had no effect on CBR3 mRNA and protein expression osteosarcoma cells.