Introduction: DNA polymerases are crucial for maintaining genome stability and influencing tumorigenesis. However, the clinical implications of DNA polymerases in tumorigenesis and their potential as anti-cancer therapy targets are not well understood. Methods: We conducted a systematic analysis using TCGA Pan-Cancer Atlas data and Gene Set Cancer Analysis results to examine the expression profiles of 15 DNA polymerases (POLYs) and their clinical correlations. We also evaluated the prognostic value of POLYs by analyzing their expression levels in relation to overall survival time (OS) using Kaplan-Meier survival curves. Additionally, we investigated the correlations between POLY expression and immune cells, DNA damage repair (DDR) pathways, and ubiquitination. Drug sensitivity analysis was performed to assess the relationship between POLY expression and drug response. Results: Our analysis revealed that 14 out of 15 POLYs exhibited significantly distinct expression patterns between tumor and normal samples across most cancer types, except for DNA nucleotidylexotransferase (DNTT). Specifically, POLD1 and POLE showed elevated expression in almost all cancers, while POLQ exhibited high expression levels in all cancer types. Some POLYs showed heightened expression in specific cancer subtypes, while others exhibited low expression. Kaplan-Meier survival curves demonstrated significant prognostic value of POLYs in multiple cancers, including PAAD, KIRC, and ACC. Cox analysis further validated these findings. Alteration patterns of POLYs varied significantly among different cancer types and were associated with poorer survival outcomes. Significant correlations were observed between the expression of POLY members and immune cells, DDR pathways, and ubiquitination. Drug sensitivity analysis indicated an inverse relationship between POLY expression and drug response. Conclusion: Our comprehensive study highlights the significant role of POLYs in cancer development and identifies them as promising prognostic and immunological biomarkers for various cancer types. Additionally, targeting POLYs therapeutically holds promise for tumor immunotherapy.