The role of microRNA deregulation in the pathogenesis of adrenocortical carcinoma
Deniz M Özata
Endocr Relat Cancer. 2011 December; 18(6): 643–655.
Adrenocortical carcinoma (ACC) is an aggressive tumor showing frequent metastatic spread and poor survival. Although recent genome-wide studies of ACC have contributed to our understanding of the disease, major challenges remain for both diagnostic and prognostic assessments. The aim of this study was to identify specific microRNAs (miRNAs) associated with malignancy and survival of ACC patients. miRNA expression profiles were determined in a series of ACC, adenoma, and normal cortices using microarray. A subset of miRNAs showed distinct expression patterns in the ACC compared with adrenal cortices and adenomas. Among others, miR-483-3p, miR-483-5p, miR-210, and miR-21 were found overexpressed, while miR-195, miR-497, and miR-1974 were underexpressed in ACC. Inhibition of miR-483-3p or miR-483-5p and overexpression of miR-195 or miR-497 reduced cell proliferation in human NCI-H295R ACC cells. In addition, downregulation of miR-483-3p, but not miR-483-5p, and increased expression of miR-195 or miR-497 led to significant induction of cell death. Protein expression of p53 upregulated modulator of apoptosis (PUMA), a potential target of miR-483-3p, was significantly decreased in ACC, and inversely correlated with miR-483-3p expression. In addition, high expression of miR-503, miR-1202, and miR-1275 were found significantly associated with shorter overall survival among patients with ACC (P values: 0.006, 0.005, and 0.042 respectively). In summary, we identified additional miRNAs associated with ACC, elucidated the functional role of four miRNAs in the pathogenesis of ACC cells, demonstrated the potential involvement of the pro-apoptotic factor PUMA (a miR-483-3p target) in adrenocortica.
The ACC cell line NCI-H295R was purchased from the American Type Culture Collection (ATCC# CRL-2128; LGC Standards, Middlesex, UK). Cells were maintained in DMEM:F12 medium containing 2.5% of NuSerum (cat. no. 355500; BD Biosciences, Bedford, MA, USA), 1% penicillin/streptomycin and 1% insulin-transferin-sodium selenite (ITS+1) liquid media supplement (cat. no. I2521; Sigma–Aldrich Logistik GmbH) at 37 °C and 5% CO2. Authentication of the cell line was evaluated and verified by Bio-Synthesis, Inc. (Lewisville, TX, USA) employing genotyping of 15 short tandem repeat (STR) loci and the amelogenin gene (AMEL), and comparison with genotype information at the ATCC (Supplementary Table 2, see section on supplementary data given at the end of this article).