Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Genome wide expression analysis in AtT-20 cells following Dnmt1 knockdown

ABSTRACT: Efficient and sustained knockdown of DNMT1 transcript and protein was achieved using a consecutive transfection protocol in the mouse pituitary adenoma cell line, AtT-20. Genome wide microarray analysis identified 91 transcripts that were significantly differentially expressed relative to cells treated with a non-targeting control. Keywords: Differential expression AtT-20 cells were repeatedly treated with 20 nM siRNA (siDNMT1 or siNT) over an eight day period. After eight days total RNA was extracted and prepared for microarray analysis. The procedure was repeated three times independently.

ORGANISM(S): Mus musculus

SUBMITTER: Kevin Dudley

PROVIDER: E-GEOD-10552 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Genome-wide analysis in a murine Dnmt1 knockdown model identifies epigenetically silenced genes in primary human pituitary tumors.

Dudley Kevin J KJ Revill Kate K Whitby Philip P Clayton Richard N RN Farrell William E WE

Molecular cancer research : MCR 20081001 10

DNA methylation at promoter CpG islands (CGI) is an epigenetic modification associated with inappropriate gene silencing in multiple tumor types. In the absence of a human pituitary tumor cell line, small interfering RNA-mediated knockdown of the maintenance methyltransferase DNA methyltransferase (cytosine 5)-1 (Dnmt1) was used in the murine pituitary adenoma cell line AtT-20. Sustained knockdown induced reexpression of the fully methylated and normally imprinted gene neuronatin (Nnat) in a tim ...[more]

PMID: 18922972

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Project description:While the hypothalamo-pituitary-adrenal axis (HPA) activates a general stress response by increasing glucocorticoid (Gc) synthesis, biological stress resulting from infections triggers the inflammatory response through production of cytokines. The pituitary gland integrates some of these signals by responding to the pro-inflammatory cytokines IL6 and LIF and to a negative Gc feedback loop. The present work used whole-genome approaches to define the LIF/STAT3 regulatory network and to delineate cross-talk between this pathway and Gc action. Genome-wide ChIP-chip identified 3 449 STAT3 binding sites, whereas 2 396 genes regulated by LIF and/or Gc were found by expression profiling. Surprisingly, LIF on its own changed expression of only 85 genes but the joint action of LIF and Gc potentiated the expression of more than a thousand genes. Accordingly, activation of both LIF and Gc pathways also potentiated STAT3 and GR recruitment to many STAT3 targets. Our analyses revealed an unexpected gene cluster that requires both stimuli for delayed activation: 83% of the genes in this cluster are involved in different cell defense mechanisms. Thus, stressors that trigger both general stress and inflammatory responses lead to activation of a stereotypic innate cellular defense response. Experiment Overall Design: 1) Pituitary corticotrophs cell line (AtT-20) response to glucocorticoids and LIF: Experiment Overall Design: AtT-20 cells mRNA extracted after treatment with PBS as control, LIF for 18h, Dex+LIF for 3h and Dex+LIF for 18h. Each treatment were made in duplicate. [GSM471246-GSM471253] Experiment Overall Design: 2) Pituitary corticotrophs cell line (AtT-20) response to LIF after 3h treatment: Experiment Overall Design: AtT-20 cells mRNA extracted after treatment with PBS as control or LIF for 3h. Each treatment were made in duplicate. [GSM471254-GSM471257] Experiment Overall Design: 3) Pituitary corticotrophs cell line (AtT-20) response to Dex after 3h treatment: Experiment Overall Design: AtT-20 cells mRNA extracted after treatment with PBS as control or Dex for 3h. Each treatment were made in duplicate. [GSM471258-GSM471261] Experiment Overall Design: 4) Pituitary corticotrophs cell line (AtT-20) response to Dex after 18h treatment: Experiment Overall Design: AtT-20 cells mRNA extracted after treatment with PBS as control or Dex for 18h. Each treatment were made in duplicate. [GSM471262-GSM471269]

Dataset Information

Genome wide expression analysis in AtT-20 cells following Dnmt1 knockdown

Publications

Genome-wide analysis in a murine Dnmt1 knockdown model identifies epigenetically silenced genes in primary human pituitary tumors.

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