Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Transcription profiling of human H295R Cells Treated with ANGII, Potassium, or Forskolin for 3 Hours

ABSTRACT: H295R human adrenocortical cells were cultured in H295R complete media containing DMEM:F12 (1:1) supplemented with 2% Ultroser G (Biosepra, Villeneuve-la-Garenne, France), ITS-Plus (Discovery Labware, Bedford, MA) and an antibiotic/antimycotic mixture (Invitrogen, Carlsbad, CA) in 175 cm2 flasks until subconfluente. Media was replaced with 40 ml fresh media containing different agents and cultured for 3 h. Treatments: Angiotensin II (100 nM), potassium (16 mM) and forskolin (10 uM). Total RNA extracted with RNAesy Midi Kit (Qiagen) with on-column DNase digestion Experiment Overall Design: Controls, n=2 Experiment Overall Design: Angiotensin=1 Experiment Overall Design: Potassium=1 Experiment Overall Design: Forskolin=1 Experiment Overall Design: Same biological sample used for GPL96 and GPL97

ORGANISM(S): Homo sapiens

SUBMITTER: Damian Gaston Romero

PROVIDER: E-GEOD-5553 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Adrenal transcription regulatory genes modulated by angiotensin II and their role in steroidogenesis.

Romero Damian G DG Rilli Silvia S Plonczynski Maria W MW Yanes Licy L LL Zhou Ming Yi MY Gomez-Sanchez Elise P EP Gomez-Sanchez Celso E CE

Physiological genomics 20070227 1

Transcription regulatory genes are crucial modulators of cell physiology and metabolism whose intracellular levels are tightly controlled to respond to extracellular stimuli. We studied transcription regulatory genes modulated by angiotensin II, one of the most important regulators of adrenal cortical cell function, and their role in adrenal steroidogenesis in H295R human adrenocortical cells. Angiotensin II-modulated transcription regulatory genes were identified with high-density oligonucleoti ...[more]

PMID: 17327493

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Project description:The mineralocorticoid aldosterone mainly produced by the adrenal gland is essential for life but an abnormal excessive secretion causes severe pathological effects including hypertension and target organ injury in the heart and kidney. The aim of this study was to determine the gene regulatory network triggered by aldosterone secretagogues in a non transformed cell system. Freshly isolated rat adrenal zona glomerulosa cells were stimulated with the two main aldosterone secretagogues, angiotensin II and potassium, for two hours and subjected to whole genome expression studies using multiple biological and bioinformatics tools. Several genes were differentially expressed by Ang II (n=133) or potassium (n=216). Genes belonging to the nucleic acid binding and transcription factor activity categories were significantly enriched. A subset of the most regulated genes were confirmed by real-time RT-PCR and then their expression analyzed in time curve studies. Differentially expressed genes were grouped according to their time-response expression pattern and their promoter regions analyzed for common regulatory transcription factors binding sites. Finally, data mining with gene promoters, transcription factors and literature databases were performed to generate gene interaction networks for either Ang II or potassium. This study provides for the first time a complete study of the genes that are regulated, and the interaction between them, by aldosterone secretagogues in rat adrenal cells. Increasing our knowledge of adrenal physiology and gene regulation in non transformed cell systems would lead us to a better approach for discovery of candidate genes involved pathological conditions of the adrenal cortex. Experiment Overall Design: Freshly isolated rat adrenal cells were treated with Ang II (100 nM) or potassium (16 mM) for 2 h at 37C. Experiment Overall Design: Total RNA from three independent experiments were isolated, labeled and hybridized to whole genome high-density oligonucleotide microarrays (Affymetrix Rat Genome 230 2.0). Experiment Overall Design: 3 independent cell preparations. Experiment Overall Design: In each experiement there is one tube control, one Ang II (100 nM) and potassium (16 mM).

Dataset Information

Transcription profiling of human H295R Cells Treated with ANGII, Potassium, or Forskolin for 3 Hours

Publications

Adrenal transcription regulatory genes modulated by angiotensin II and their role in steroidogenesis.

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