Objective <p>Zinc finger protein 367 (ZNF367) acts as a transcription factor and has been identified as overexpressed in numerous cancers. However, the clinical significance of ZNF367 in various cancers remains fragmentary. Therefore, this study performs a comprehensive pan-cancer analysis of ZNF367 for its diagnostic and prognostic value and underlying biological mechanisms across multiple cancer types.</p> Methods <p>Multi-omics data from the website databases were utilized for analyzing the pan-cancer expression landscape of ZNF367 and its potential diagnostic and prognostic significance, and validating its expression in some cancers by immunohistochemistry, followed by a series of biological analyses using online bioinformatics tools to investigate the underlying mechanisms, including the genetic and epigenetic alterations of the ZNF367 gene, interaction networks of ZNF367 with proteins and genes, associations of ZNF367 with tumor genetic heterogeneity and tumor microenvironment characteristics.</p> Results <p>ZNF367 expression levels were elevated in the majority of cancers, validated its overexpression of protein in some cancers, and showed moderate to high diagnostic performance in 26 cancer types. Overexpression of ZNF367 was connected with unfavorable prognostic outcomes across several malignancies and acted as an independent risk factor using multivariate Cox regression analysis in some tumors. Common genetic alterations of ZNF367 included mutations and amplifications. KEGG functional analysis uncovered that ZNF367-associated genes participated in multiple cancer-related pathways. Significant correlations were observed between ZNF367 and tumor genetic heterogeneity, DNA methylation, immune infiltration and ZNF367-related genes in most cancers.</p> Conclusion <p>ZNF367 may emerge as a valuable pan-cancer diagnostic and prognostic biomarker supported by biological evidence.</p>