Diagnosis of malignancy in adrenocortical tumors can be challenging for pathologists and prognostic of adrenocortical carcinoma (ACC) remains difficult to predict. MiR-483-5p and miR-139-5p were shown to be upregulated in ACC compared with adrenocortical adenoma (ACA), and N-myc Downstream-Regulated Gene 2 (NDRG2) and NDRG4 were identified as targets of miR-483-5p and miR-139-5p, respectively. In this study, we investigated NDRG2 and NDRG4 expression using immunohistochemistry (IHC), as well as miR-483-5p and miR-139-5p expression using in situ hybridization (ISH), by a H-score-like method on formalin-fixed and paraffin-embedded adrenocortical tissue sections. These semi-quantitative approaches were correlated with mRNA and miR expression measured by Real Time quantitative PCR (RT-qPCR) and protein levels determined by western blot in frozen tissues. Ten ACA and twenty ACC were retrospectively selected. ACC were divided into two groups, one consisting of aggressive ACC (aACC) as opposed to non-aggressive ACC (naACC). NDRG2 displayed a significant lower expression in ACC compared with ACA (p<0.001) and a score of 152.5 was determined as the cutoff value which best discriminated ACA from ACC with a sensitivity of 100% and a specificity of 95%. NDRG4 expression under 35 allowed to discriminate between aACC and naACC with a sensitivity of 100% and a specificity of 75%. A strong correlation was observed between the scores determined by IHC and their expression measured by RT-qPCR and protein levels determined by western blot. In conclusion, our study demonstrates that NDRG2 and NDRG4 are potential useful immunomarkers for ACC diagnosis and prognosis.