Furthermore, gene set enrichment analysis (GSEA) with Gene Expression Omnibus (GEO) E-MEXP-3628 dataset showed that TRIM66 was positive related with apoptosis, epithelial-mesenchymal transition (EMT) and transforming growth factor-β (TGF-β) signaling pathway, which was further confirmed in osteosarcoma cells with TRIM66 silenced. This evidence concerns the gene TRIM66 and osteosarcoma.