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Gene regulation in ki-ras transfected murine fibroblasts

ABSTRACT: Ki-ras gene mutations are involved in the carcinogenesis of acute myeloid leukemia, melanoma and different carcinomas. In order to define potential mutation-specific therapeutic targets, expression profiling analyses were performed using stable transfected NIH3T3 cells carrying different ki-ras gene mutations. Maybe, this analysis allows the discorvery of ras-associated cellular mechanisms, which might lead to the identification of physiological targets for pharmacological interventions of the treatment of Ki-ras-associated human tumors. Keywords: cDNA microarray, murine fibroblast, ki-ras mutations, carcinogenesis 4 transfected murine fibroblast cell lines (mock, wt-kras, Asp, Val) were compared to non-transfected cell line. For each of the four comparison 4 chip experiments were performed including 2 dye swaps.

ORGANISM(S): Mus musculus

SUBMITTER: Johannes Beckers

PROVIDER: E-GEOD-8372 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Overexpressed vs mutated Kras in murine fibroblasts: a molecular phenotyping study.

Horsch M M Recktenwald C V CV Schädler S S Hrabé de Angelis M M Seliger B B Beckers J J

British journal of cancer 20090203 4

Ras acts in signalling pathways regulating the activity of multiple cellular functions including cell proliferation, differentiation, and apoptosis. Amino-acid exchanges at position 12, 13, or 61 of the Kras gene convert the proto-oncogene into an activated oncogene. Until now, a direct comparison of genome-wide expression profiling studies of Kras overexpression and different Kras mutant forms in a single assay system has not been carried out. In our study, we focused on the direct comparison o ...[more]

PMID: 19190631

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Gene regulation in Erbb2 transfected NIH3T3 cells after Azacytidine treatment

Project description:5-Azacytidine or 5-aza-2â??-deoxycytidine is a chemical analogue of the cytosine nucleoside used in DNA and RNA. Cells in the presence of 5-azacytidine incorporate it into DNA during replication and RNA during transcription. The incorporation of 5-azacytidine into DNA or RNA inhibits methyltransferase thereby causing demethylation in that sequence, affecting the way that cell regulation proteins are able to bind to the DNA/RNA substrate. Inhibition of DNA occurs through the formation of stable complexes between the molecule and with DNA methyltransferases, thereby saturating the cells methylation machinery. 5-azacytidine is mainly used in the treatment of myelodysplastic syndrome. Changes in gene expression levels in Erbb2 transfected murine fibroblast cell line after treatment with Azacytidine in two different concentrations were analysed. Keywords: cDNA microarray, murine fibroblast, azacytidine treatment, carcinogenesis Gene expression levels of Erbb2 transfected NIH3T3 cell lines treated with 5ÂµM and 10ÂµM Azacytidine were compared to Erbb2 transfected but non-treated NIH3T3 cell line. For each Azacytidine concentration 4-8 experiments were performed including 50% dye swaps. As a controll experiments gene expression levels of NIH3T3 cells with and without Azacytidine treatment were analysed

2009-10-01 | E-GEOD-8374 | biostudies-arrayexpress

Gene regulation in brain, eye and heart of sighted C3H mice

Project description:It is unclear what role vision plays in guiding mouse behaviour, since the mouse eye is of comparably low optical quality, and mice are considered to rely primarily on other senses. All C3H substrains are homozygous for the Pde6b(rd1) mutation and get blind by weaning age. To study the impact of the Pde6b(rd1) mutation on mouse behaviour and physiology, sighted C3H (C3H.Pde6b+) and normal C3H/HeH mice were phenotyped for different aspects. We confirmed retinal degeneration 1 in C3H/HeH mice, and the presence of a morphologically normal retina as well as visual ability in C3H.Pde6b+ mice. However, C3H.Pde6b+ mice showed an abnormal retinal function in the electroretinogram response, indicating that their vision was not normal as expected. C3H.Pde6b+ mice showed reduced latencies for several behaviours without any further alterations in these behaviours in comparison to C3H/HeH mice, suggesting that visual ability, although impaired, enables earlier usage of the behavioural repertoire in a novel environment, but does not lead to increased activity levels. These results emphasize the importance of comprehensive behavioural and physiological phenotyping. RNA expression profiling of eye, brain and heart of six male animals of the C3H seh mutant mouse line. In total 32 chip hybridizations including biological and experimental replicates (colour flip) were performed. The data analysis and various statistical methods detected several genes differentially regulated between mutant and reference tissues in all analyzed organ types.

2010-05-23 | E-GEOD-7855 | biostudies-arrayexpress

Gene expression profile of a murin neuroblastoma cell line infected with the prion strain 22L

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2010-05-26 | E-GEOD-7119 | biostudies-arrayexpress

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2010-06-21 | E-GEOD-6672 | biostudies-arrayexpress

Ovalbumin sensitized and challenged induces differential gene expression in mouse lungs

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2010-06-10 | E-GEOD-6496 | biostudies-arrayexpress

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2010-05-22 | E-GEOD-9065 | biostudies-arrayexpress

UCP exposure of ovalbumin sensitized and challenged mice

Project description:To gain insight into the promoting effect of ultrafine particle inhalation on development and progression of allergic asthma, we selected an experimental approach involving exposure to ultrafine carbon particles (UCP) and gene expression profiling of lungs from mice with experimental, ovalbumin induced allergy. Comparative gene expression analysis was performed by hybridizing pooled cDNA samples from lavaged lungs of different groups. The results suggest that allergic sensitization may represent an susceptibility factor for effects of UCP on gene expression in the lung. In sensitized individuals UCP exposure, such as found in polluted air, thus may contribute to the development and/or aggrevation of allergic asthma. Keywords: Particle Inhalation, lung, ovalbumin sensitized and challanged, expression profling Lungs of groups of six sensitized or sensitized and challanged BALB/cJ mice, either subjected to particle-free or UCP containing air; two replicates including one dye swap experiment have been performed for lungs: a) sensitized particle free air versus sensitized UCP exposure; b) sensitized and challanged particle free air versus sensitized and challanged UCP exposure

2010-06-10 | E-GEOD-6571 | biostudies-arrayexpress

Identification and validation of novel ERBB2 (HER2, NEU) targets including genes

Project description:V-erb-b2 erythroblastic leukemia viral oncogene homolog 2 (ERBB2; synonyms HER2, NEU) encodes a transmembrane glycoprotein with tyrosine kinase-specific activity that acts as a major switch in different signal-transduction processes. ERBB2 amplification and overexpression have been found in a number of human cancers, including breast, ovary and kidney carcinoma. Our aim was to detect ERBB2- regulated target genes that contribute to its tumorigenic effect on a genomewide scale. The differential gene expression profile of ERBB2- transfected and wild-type mouse fibroblasts was monitored employing DNA microarrays. Regulated expression of selected genes was verified by RT-PCR and validated by Western blot analysis. Genomewide gene expression profiling identified (i) known targets of ERBB2 signaling, (ii) genes implicated in tumorigenesis but so far not associated with ERBB2 signaling as well as (iii) genes not yet associated with oncogenic transformation, including novel genes without functional annotation. We also found that at least a fraction of coexpressed genes are closely linked on the genome. ERBB2 overexpression suppresses the transcription of antiangiogenic factors (e.g., Sparc, Timp3, Serpinf1) but induces expression of angiogenic factors (e.g., Klf5, Tnfaip2, Sema3c). Profiling of ERBB2-dependent gene regulation revealed a compendium of potential diagnostic markers and putative therapeutic targets. Identification of coexpressed genes that colocalize in the genome may indicate gene regulatory mechanisms that require further study to evaluate functional coregulation. Erbb2 transfected NIH3T3 cells versus un-transfected cells, 4 technical replicates including two colour swap experiments

2010-06-25 | E-GEOD-16445 | biostudies-arrayexpress

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2010-05-19 | E-GEOD-15336 | biostudies-arrayexpress

MFAP4 is an integrin M-NM-1VM-NM-23/5 ligand involved in vascular remodeling

Project description:Arterial injury or occlusive arterial disease may stimulate healing responses, which when overexuberant, leads to restenosis of the injured vessel. This response is influenced by specific integrin signalling in vascular smooth muscle cells (VSMCs). Microfibril-associated protein 4 (MFAP4) colocates in blood vessels with elastic fibers. MFAP4 contains an N-terminal RGD-motiv, which is a potential integrin binding site. The role for MFAP4 in vascularproliferative disease is so far unknown and the subject for these investigations. Here we show that MFAP4 is expressed and secreted by VSMCs and binds elastin and collagen. MFAP4 mediated adhesion, and migration, and proliferation of VSMCs in an integrin M-NM-1VM-NM-23/5 dependent manner and the effects were inhibited by MFAP4 blocking antibodies. MFAP4 deficient mutant mice were generated and appeared with a normal cardiophysiological phenotype. When challenged by carotid artery ligation, the MFAP4 deficient mice had delayed neointimal formation and the compensatory outward remodeling of the vessel diameter and thus the vessel lumen was reduced. MFAP4 expression appeared unaffected by the induced pathology. HUMANE DATA This new MFAP4 mediated molecular mechanism for regulation of integrin M-NM-1VM-NM-23/5 signalling may have therapeutic implications in diseases where VSMC migration and proliferation are involved in the pathogenesis. For genome-wide expression analysis total RNA from heart of four male MFAP4-/- and four control mice was isolated using RNeasy Midi kit (Qiagen, Hilden, Germany). The cDNA microarrays were generated, hybridized and analysed as described (Horsch et al 2009). Two chip hybridizations were performed with total RNA for each individual mutant mouse against a reference RNA pool of the same organ.

2013-04-01 | E-GEOD-40480 | biostudies-arrayexpress

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