Background Colorectal cancer (CRC) is, at present, the most frequently and most lethal gastrointestinal tract cancer, leading to significant mortality and a large global cancer burden. TSPEAR has been reported to associate with the development of human tooth and hair follicle morphogenesis. However, there is no study on TSPEAR in any cancers have been reported. Methods The information of TSPEAR gene expression in CRC was collected from TCGA database and GTEx. The Rt-qPCR and WB were used to compare the TSPEAR expression level between normal colon cells NCM-460 and human CRC cell lines, included SW-480, SW-620, HCT-116, Ht-29, and LoVo. Then, we performed the survival, gene function/pathway enrichment, and tumor immune infiltration analyses via using R packages and TIMER database. Results It was demonstrated that TSPEAR gene showed high expression in CRC tissue and cell lines and up-regulated expression correlating with a poor overall prognosis. Besides, the up-regulated TSPEAR was correlated with the M1 stage, CEA level, and residual tumor. Multivariate analysis further confirmed that TSPEAR remained an independent CRC patient-related risk factors, whereas gene function/pathway enrichment analysis indicated that it may can regulate Wnt, Hippo signaling pathway, as well as many cell metabolism activities. For immune infiltration, the expression level of TSPEAR was positively correlated with immune cells and showed a strong correlation with diverse immune marker sets, especially in COAD. Conclusions The results demonstrated that TSPEAR is possibly a clinically valuable predictive biomarkers for CRC patients, as well as associated with tumor infiltrating immune cells.