Purkinje Cell Protein 4 Expression Is Associated With DNA Methylation Status in Aldosterone-Producing Adenoma
| Autor: | Kenji Oki, Kazuhiro Kobuke, Masayasu Yoneda, Celso E. Gomez-Sanchez, Haruya Ohno, Noboru Hattori, Kiyotaka Itcho, Yoko Yoshii |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Adult Male medicine.medical_specialty Genotype Endocrinology Diabetes and Metabolism Clinical Biochemistry Nerve Tissue Proteins Biology Methylation Site Biochemistry 03 medical and health sciences Endocrinology Calmodulin Transcription (biology) Enhancer binding Internal medicine CEBPA medicine Tumor Cells Cultured Humans Epigenetics Calcium Signaling RNA Messenger RNA Neoplasm Promoter Regions Genetic Aldosterone Biochemistry (medical) Methylation DNA Neoplasm DNA Methylation Middle Aged Molecular biology Adrenal Cortex Neoplasms Gene Expression Regulation Neoplastic 030104 developmental biology DNA methylation Adrenocortical Adenoma Mutation Female Chromatin immunoprecipitation |
| Zdroj: | The Journal of clinical endocrinology and metabolism. 103(3) |
| ISSN: | 1945-7197 |
| Popis: | Context Aldosterone production is stimulated by activation of calcium signaling in aldosterone-producing adenomas (APAs), and epigenetic factors such as DNA methylation may be associated with the expression of genes involved in aldosterone regulation. Objective Our aim was to investigate the DNA methylation of genes related to calcium signaling cascades in APAs and the association of mutations in genes linked to APAs with DNA methylation levels. Methods Nonfunctioning adrenocortical adenoma (n = 12) and APA (n = 35) samples were analyzed. The KCNJ5 T158A mutation was introduced into human adrenocortical cell lines (HAC15 cells) using lentiviral delivery. DNA methylation array analysis was conducted using adrenal tumor samples and HAC15 cells. Results The Purkinje cell protein 4 (PCP4) gene was one of the most hypomethylated in APAs. DNA methylation levels in two sites of PCP4 showed a significant inverse correlation with messenger RNA expression in adrenal tumors. Bioinformatics and multiple regression analysis revealed that CCAAT/enhancer binding protein alpha (CEBPA) may bind to the methylation site of the PCP4 promoter. According to chromatin immunoprecipitation assay, CEBPA was bound to the PCP4 hypomethylated region by chromatin immunoprecipitation assay. There were no significant differences in PCP4 methylation levels among APA genotypes. Moreover, KCNJ5 T158A did not influence PCP4 methylation levels in HAC15 cells. Conclusions We showed that the PCP4 promoter was one of the most hypomethylated in APAs and that PCP4 transcription may be associated with demethylation as well as with CEBPA in APAs. KCNJ5 mutations known to result in aldosterone overproduction were not related to PCP4 methylation in either clinical or in vitro studies. |
| Databáze: | OpenAIRE |
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