Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

RNAi knock down of RET finger protein/ Tripartite protein in human osteosarcoma cell lines and analysis of Retinoblastoma protein dependent transcriptional changes

ABSTRACT: Analysis of Retinoblastoma protein (Rb) (Hs.408528) dependent transcriptional changes following siRNA mediated ablation of the RET finger protein/ Tripartite protein (RFP/TRIM27) (Hs.440382). Common reference design, two biological replicates with two technical replicates each.

ORGANISM(S): Homo sapiens

SUBMITTER: Sibylle Mittnacht

PROVIDER: E-MEXP-278 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Selective ablation of retinoblastoma protein function by the RET finger protein.

Krützfeldt Maja M Ellis Mark M Weekes Daniel B DB Bull Jonathan J JJ Eilers Martin M Vivanco Maria D M MD Sellers William R WR Mittnacht Sibylle S

Molecular cell 20050401 2

The retinoblastoma tumor suppressor protein (Rb) affects gene transcription both negatively and positively and through this regulates distinct cellular responses. Although cell cycle regulation requires gene repression, Rb's ability to promote differentiation and part of its antiproliferative activity appears to rely on the activation of gene transcription. We present evidence here that the RET finger protein (RFP)/tripartite motif protein 27 (TRIM 27) inhibits gene transcription activation by R ...[more]

PMID: 15837424

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Project description:In addition to their role in the development and function of the reproductive system, estrogens have significant anti-inflammatory properties. Although both estrogen receptors (ERs) can mediate anti-inflammatory actions, ERbeta is a more desirable therapeutic target because ERalpha mediates the proliferative effects of estrogens on the mammary gland and uterus. In fact, selective ERbeta agonists have beneficial effects in preclinical models involving inflammation without causing growth-promoting effects on the uterus or mammary gland. However, their mechanism of action is unclear. The purpose of this study was to use microarray analysis to determine whether ERbeta-selective compounds produce their anti-inflammatory effects by repressing transcription of proinflammatory genes. We identified 49 genes that were activated by TNF-alpha in human osteosarcoma U2OS cells expressing ERbeta. Estradiol treatment significantly reduced the activation by TNF-alpha on 18 genes via ERbeta or ERalpha. Most repressed genes were inflammatory genes, such as TNF-alpha, IL-6, and CSF2. Three ERbeta-selective compounds, ERB-041, WAY-202196, and WAY-214156, repressed the expression of these and other inflammatory genes. ERB-041 was the most ERbeta-selective compound, whereas WAY-202196 and WAY-214156 were the most potent. The ERbeta-selective compounds repressed inflammatory genes by recruiting the coactivator, SRC-2. ERB-041 also repressed cytokine genes in PBMCs, demonstrating that ERbeta-selective estrogens have anti-inflammatory properties in immune cells. Our study suggests that the anti-inflammatory effects of ERB-041 and other ERbeta-selective estrogens in animal models are due to transcriptional repression of proinflammatory genes. These compounds might represent a new class of drugs to treat inflammatory disorders. Experiment Overall Design: Expression of ER in the U2OS-ERÃ cells was induced with doxycycline. The cells were treated in the absence or presence of 10 nM E2 for 18 h followed by incubation with TNF-{alpha} for 1 h. Total RNA were extracted, followed by cRNA labelling, and hybridization of fragmented samples to the U95Av2 arrays (n = 3 for untreated, n = 4 for TNF-{alpha}-treated, n = 4 for TNF-{alpha}- and E2-treated samples). The data were analyzed using the Microarray Suite Version 5.0 with the default parameters. The comparative data generated for each treated group were analyzed further in Microsoft Excel. TNF-{alpha}-induced genes were selected for further analysis only if they had a 1.2 signal log ratio mean value (2.3-fold change) and were statistically significant (p < 0.05) in at least three separate experiments.

Dataset Information

RNAi knock down of RET finger protein/ Tripartite protein in human osteosarcoma cell lines and analysis of Retinoblastoma protein dependent transcriptional changes

Publications

Selective ablation of retinoblastoma protein function by the RET finger protein.

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