Abstract
DTX3L plays pivotal roles in diverse biological processes, yet its multifaceted functions and the molecular mechanisms driving tumorigenesis and cancer progression remain incompletely defined. Here, we systematically investigated DTX3L’s expression profiles, prognostic value, and potential regulatory mechanisms across pan-cancers. By integrating data from multiple databases, we analyzed the expression levels of DTX3L and its correlations with clinicopathological features, prognosis, and immune infiltration. Leveraging the STRING database, we identified DTX3L-associated genes and performed functional enrichment analysis. Finally, we experimentally validated the impact of DTX3L knockdown on the proliferation and migration of ovarian cancer cells. We found DTX3L was significantly upregulated in most human tumors, with its expression correlating with patient age, TNM stage, grade, and clinical stage—indicating DTX3L as a biomarker of poor prognosis. DTX3L showed extensive co-expression with key RNA modification genes and correlations with immune checkpoint genes, immune cells, and immune infiltration scores. GO and KEGG analyses linked DTX3L to DNA damage repair, ubiquitination regulation, and innate immune signaling pathways. In ovarian cancer cells, DTX3L knockdown markedly reduced cell proliferation, migration, and increased apoptosis. These findings demonstrate that DTX3L is upregulated in various cancers and is associated with poor patient prognosis, suggesting its potential as both a prognostic biomarker and a therapeutic target in pan-cancer immunotherapy. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1038/s41598-025-32552-3.