Project description:Purpose: The management of adrenocortical tumors (ACTs) is complex, compounded by the difficulty in discriminating benign from malignant tumors using conventional histology. The Weiss score is the current most widely used system for ACT diagnosis but it has limitations, particularly with ACTs with a score of 3. The am of this study was to identify molecular markers whose expression can discriminate adrenocortical carcinomas (ACCs) from adrenocortical adenomas (ACAs) by microarray gene expression profiling and to determine their clinical applicability by using immunohistochemistry (IHC). Experimental design: Microarray gene expression profiling was used to identify 7 molecular markers which were significantly differentially expressed between ACCs and ACAs. These results were confirmed with quantitative PCR for all 7 genes and IHC for 3 protein. Results: Microarray gene expression profiling was able to accurately categorize ACTs into ACCs and ACAs. All 7 genes were strong discriminators of ACCs from ACAs on qPCR. IHC with IGF2, MAD2L1, CCNB1 and Ki-67, but not ACADVL or ALOX15B, had high diagnostic accuracy in differentiating ACCs from ACAs. The best results however were obtained with a combination of IGF2 and Ki-67 with 96% sensitivity and 100% specificity in diagnosing ACCs. Conclusion: Microarray gene expression profiling accurately differentiates ACCs from ACAs. The combination of IGF2 and Ki-67 IHC is also highly accurate in distinguishing between the 2 groups and is particularly helpful in ACTs with Weiss score of 3. Keywords: Adrenocortical carcinoma, adrenocortical adenoma, differential gene expression, immunohistochemistry, qPCR
Project description:Mutations of β-catenin gene (CTNNB1) are frequent in adrenocortical adenomas (AA) and carcinomas (ACC). However, the target genes of CTNNB1 have not yet been identified in adrenocortical tumors. Our objective was to identify genes de-regulated in adrenocortical tumors harbouring CTNNB1 genetic alterations.
Project description:Genome wide DNA methylation profiling of normal adrenocortical tissue, adrenocortical adenomas and adrenocortical carcinomas. The Illumina Infinium 27k Human DNA methylation Beadchip v1.2 was used to obtain DNA methylation profiles. Samples included 6 normal adrenocortical tissue samples, 27 adenomas and 15 carcinomas.
Project description:Mutations of β-catenin gene (CTNNB1) are frequent in adrenocortical adenomas (AA) and carcinomas (ACC). However, the target genes of CTNNB1 have not yet been identified in adrenocortical tumors. Our objective was to identify genes de-regulated in adrenocortical tumors harbouring CTNNB1 genetic alterations. We compared gene expression profiles of AA with (n: 3) and without (n: 4) CTNNB1 mutations using Affymetrix arrays.
Project description:Micro-RNA sequencing of adrenocortical tumors and normal adrenal samples. miRNA sequencing of 45 adrenocortical carcinomas (ACC), 30 adrenocortical adenomas (ACA) and 3 normal adrenal samples.
Project description:MicroRNAs are small, non-coding RNAs that regulate gene expression at post-transcriptional levels. There is increasing evidence to suggest that miRNAs could be useful in cancer diagnosis, prognosis, and therapy. The aim of our study was to identify miRNAs predictors of poor prognosis in adrenocortical cancer. miRNA microarray expression profiling was performed on a cohort of 6 adenomas, 6 non-recurrent carcinomas (Carc_B) and 6 recurrent carcinomas (Carc_A). We identified several miRNAs that were differentially expressed between adenomas and carcinomas as well as between Carc_A and Carc_B. We found that the best discriminatory miRNAs between carcinomas and adenomas were miR-195 and miR-335 which were down-regulated in carcinomas. MiR-139-5p was the most powerful discriminatory miRNA between Carc_A and Carc_B subtypes with consistent up-regulation in the recurrent carcinoma subgroup (Carc_A). Target prediction analysis showed that predicted targets of these miRNAs could be involved in biological processes and pathways that enhance tumor progression. Our data suggest that adrenocortical cancer cells progressively switch from a high miR-195 and miR-335 status to a low miR-195 and miR-335 phenotype. MiR-139-5P is a potential prognostic biomarker of recurrent adrenocortical carcinomas.
Project description:Pediatric adrenocortical tumors (ACT) are rare and often fatal malignancies; little is known regarding their etiology and biology. To provide additional insight into the nature of ACT, we determined the gene expression profiles of 24 pediatric tumors (five adenomas, 18 carcinomas, and one undetermined) and seven normal adrenal glands. Distinct patterns of gene expression, validated by quantitative real-time PCR and Western blot analysis, were identified that distinguish normal adrenal cortex from tumor. Differences in gene expression were also identified between adrenocortical adenomas and carcinomas. In addition, pediatric adrenocortical carcinomas were found to share similar patterns of gene expression when compared with those published for adult ACT. This study represents the first microarray analysis of childhood ACT. Our findings lay the groundwork for establishing gene expression profiles that may aid in the diagnosis and prognosis of pediatric ACT, and in the identification of signaling pathways that contribute to this disease. We used microarrays to explore the expression profiles differentially expressed in childhood adrenocortical tumors and in normal adrenal gland tissues. Pediatric adrenocortical adenoma and carcinoma patients were enrolled on the International Pediatric Adrenocortical Tumor Registry (IPACTR) and Bank protocol. Tumor specimens were harvested during surgery and snap frozen in liquid nitrogen to preserve tissue integrity. Data have been compiled for eight males and 15 females between 0 and 16 years of age. Table 1 (West et al, Cancer Research 67:601-608, 2007) summarizes the primary clinical information for each subject (excluding sample Unk1 with ACT of undetermined histology), including stage of the disease, tumor class, sex, age, relapse-free survival, and overall survival.
Project description:MicroRNAs are small, non-coding RNAs that regulate gene expression at post-transcriptional levels. There is increasing evidence to suggest that miRNAs could be useful in cancer diagnosis, prognosis, and therapy. The aim of our study was to identify miRNAs predictors of poor prognosis in adrenocortical cancer. miRNA microarray expression profiling was performed on a cohort of 6 adenomas, 6 non-recurrent carcinomas (Carc_B) and 6 recurrent carcinomas (Carc_A). We identified several miRNAs that were differentially expressed between adenomas and carcinomas as well as between Carc_A and Carc_B. We found that the best discriminatory miRNAs between carcinomas and adenomas were miR-195 and miR-335 which were down-regulated in carcinomas. MiR-139-5p was the most powerful discriminatory miRNA between Carc_A and Carc_B subtypes with consistent up-regulation in the recurrent carcinoma subgroup (Carc_A). Target prediction analysis showed that predicted targets of these miRNAs could be involved in biological processes and pathways that enhance tumor progression. Our data suggest that adrenocortical cancer cells progressively switch from a high miR-195 and miR-335 status to a low miR-195 and miR-335 phenotype. MiR-139-5P is a potential prognostic biomarker of recurrent adrenocortical carcinomas. Six tumor samples from patients with adrenocortical adenomas (Adenoma_1 to Adenoma_6), six tumor samples from patients with aggressive carcinomas (Carc_A1 to Carc_A6) and six tumor samples from patients with non-aggressive carcinoma (Carc_B1 to Carc_B6) were used to prepare total RNA for microarray analysis using MiRXploreTM Microarrays. miRXplore Universal Reference was used as control. One tumor sample was used per array.
Project description:Pediatric adrenocortical tumors (ACT) are rare and often fatal malignancies; little is known regarding their etiology and biology. To provide additional insight into the nature of ACT, we determined the gene expression profiles of 24 pediatric tumors (five adenomas, 18 carcinomas, and one undetermined) and seven normal adrenal glands. Distinct patterns of gene expression, validated by quantitative real-time PCR and Western blot analysis, were identified that distinguish normal adrenal cortex from tumor. Differences in gene expression were also identified between adrenocortical adenomas and carcinomas. In addition, pediatric adrenocortical carcinomas were found to share similar patterns of gene expression when compared with those published for adult ACT. This study represents the first microarray analysis of childhood ACT. Our findings lay the groundwork for establishing gene expression profiles that may aid in the diagnosis and prognosis of pediatric ACT, and in the identification of signaling pathways that contribute to this disease.
Project description:We performed miRNA expression profiling in a series of adrenocortical carcinomas, adrenocortical adenomas and normal adreno cortex using a microarray approach. Significant differentially expressed miRNAs among groups were identified using SAM analysis. Agilent microarray platform containing 903 miRNAs was used to determine miRNA expression profiles in 4 normal cortices, 26 adenomas and 22 adrenocortical carcinomas. SAM analysis was adopted to identify significant differentially expressed miRNAs between groups. SAM survival analysis was used to determine the association between miRNA expression and survival among carcinoma cases. The expression levels of candidate prognostic miRNAs were evaluated using qRT-PCR in the same cohort of adrenocortical carcinomas and association with survival was evaluated using Kaplan Meier curves and log rank tests.