Immune, metabolic, and steroidogenesis-associated profiles of miR-940, miR-375, and miR-326 in adrenocortical carcinoma.

Adrenocortical carcinoma (ACC) has remained a highly aggressive endocrine malignancy characterized by extensive molecular heterogeneity and limited therapeutic options. In the present study, the regulatory significance of hsa-miR-940, hsa-miR-375, and hsa-miR-326, previously identified as central hubs within an ACC competing endogenous RNA (ceRNA) network, was comprehensively characterized through integrative transcriptomic, prognostic, and pathway-level analyses using TCGA-ACC tumor samples. Network topology analysis, target prediction, survival modeling, pathway enrichment, protein-protein interaction analysis, therapeutic-response evaluation, and pan-cancer expression characterization were subsequently performed to investigate the biological relevance of these central miRNAs. Distinct network-topology differences were identified between tumor and normal ceRNA networks, with hsa-miR-940 demonstrating increased tumor-associated regulatory centrality, whereas hsa-miR-326 exhibited comparatively higher centrality within normal adrenal networks. Survival analyses further demonstrated that elevated expression of the combined three-miRNA signature was associated with significantly reduced overall survival in patients with ACC (HR = 3.52, p = 1.36E-03). Functional enrichment analyses revealed extensive associations with complement activation, steroidogenic regulation, mitochondrial bioenergetics, extracellular matrix remodeling, immune-associated signaling, lipid metabolism, and apoptosis-related pathways. Protein-protein interaction analysis additionally identified highly interconnected immune- and metabolism-associated hub genes, including CCL2, C3, SERPING1, PPARGC1A, and CFH, within the hsa-miR-940 regulatory network. Cross-cancer analyses further demonstrated substantial context-dependent expression heterogeneity across tumor types. The integrative findings presented in this study support the biological plausibility that hsa-miR-940, hsa-miR-375, and hsa-miR-326 may participate in coordinated regulatory programs associated with ACC progression and endocrine tumor adaptation. Insight box This study explores how three small regulatory molecules, known as microRNAs (miR-940, miR-375, and miR-326), may influence the development and progression of adrenocortical carcinoma (ACC), a rare and aggressive adrenal cancer. By analyzing large-scale gene expression data from tumor and normal adrenal tissues, important differences were identified in immune-related, metabolic, and hormone-associated biological processes linked to these microRNAs. The findings suggest that these molecules may contribute to how ACC tumors grow, adapt, and interact with their surrounding environment. This work demonstrates how integrative computational and biological analyses can help uncover potential biomarkers and future therapeutic targets in rare cancers such as ACC.