ELAVL1-KO Affects Steroid Synthesis in ACC Cell Line NCI-H295R and Reduces Colony-Forming Abilities

Adrenocortical carcinoma (ACC) is a rare malignancy of the adrenal gland for which no curative treatment options exist in advanced stages. Analyzing the ACC cohort of The Cancer Genome Atlas, we identified that expression of the mRNA-stabilizing protein Embryonic-Lethal-Abnormal-Vision-Like RNA-Binding Protein 1 (ELAVL1) was negatively associated with patient survival. To investigate the functional role in ACC, we generated a CRISPR/Cas9-mediated ELAVL1 knockout (KO) in the ACC cell lines NCI-H295R and HAC15 and performed Next-Generation RNA Sequencing (NGS) to characterize transcriptomic changes. In NCI-H295R, NGS analysis revealed 3468 upregulated genes and 3458 downregulated genes in ELAVL1-deficient cells compared with controls. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis identified significant enrichment of aldosterone biosynthesis-related genes and enriched downregulation of genes associated in with "pathways in cancer." Functionally, ELAVL1-KO cells exhibited impaired colony-forming ability, indicating reduced proliferative or clonogenic potential. Steroid profiling of cell culture supernatants further demonstrated increased aldosterone synthesis and decreased cortisol and androgen production in the KO cells, consistent with the observed transcriptional changes. Given that hypercortisolism is an established negative prognostic factor in ACC, these data suggest that ELAVL1 may represent a potential therapeutic target worth further investigation for modulating steroidogenesis in ACC patients.