It is well-established that SPIB is essential for the survival of mature B cells, playing a key role in diffuse large B-cell lymphoma, colorectal cancer, and lung cancer. However, no study has hitherto conducted a systematic pan-cancer analysis on SPIB. Herein, we analyzed the differential expression of SPIB in pan-cancer using The Cancer Genome Atlas (TCGA) and Genotype-Tissue Expression (GTEx) databases and found that SPIB was significantly upregulated in most cancers. In addition, SPIB was positively or negatively associated with prognosis in different cancers. We found that SPIB was significantly associated with tumor immune infiltration and immune checkpoint genes in more than 35 tumors by TIMER database analysis. In addition, SPIB was negatively correlated with Tumor mutational burden (TMB) and Microsatellite instability (MSI) in most tumors. Finally, GO/KEGG enrichment analysis revealed the possible involvement of SPIB in NF-kappa B and B-cell receptor signaling pathways. In conclusion, our comprehensive pan-cancer analysis of SPIB reveals its important role in tumor immunity, suggesting it has huge prospects for clinical application in cancer therapy.