ATM Requirement in Gene Expression Responses to Ionizing Radiation in Human Lymphoblasts and Fibroblasts
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
SUBMITTER: Kristina Flores
PROVIDER: E-GEOD-3174 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
Publications
ATM requirement in gene expression responses to ionizing radiation in human lymphoblasts and fibroblasts.
Innes Cynthia L CL Heinloth Alexandra N AN Flores Kristina G KG Sieber Stella O SO Deming Paula B PB Bushel Pierre R PR Kaufmann William K WK Paules Richard S RS
Molecular cancer research : MCR 20060301 3
The heritable disorder ataxia telangiectasia (AT) is caused by mutations in the AT-mutated (ATM) gene with manifestations that include predisposition to lymphoproliferative cancers and hypersensitivity to ionizing radiation (IR). We investigated gene expression changes in response to IR in human lymphoblasts and fibroblasts from seven normal and seven AT-affected individuals. Both cell types displayed ATM-dependent gene expression changes after IR, with some responses shared and some responses v ...[more]
Similar Datasets
Project description:This study examined the renal gene expression profiles between spontaneously-hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats at a pre-hypertensive stage (3 weeks of age) and hypertensive stage (9 weeks of age). In addition, age-related changes in gene expression patterns were examined from 3 to 9 weeks in both WKY and SHR. Five to six individual kidney samples of the same experimental group were pooled together and quadruplicate hybridizations were performed on the NIEHS Rat v2.1, 2.2 and 3.0 arrays.
2010-06-05 | E-GEOD-1932 | biostudies-arrayexpress
Project description:The parent S. cerevisiae strain, BY4741, and specific gene deletions of the parent S.cerevisiae strain BY4741 were treated with 100uM NaAsO2 (AsIII) for 2h and compared to the untreated counterpart (yap1Î? vs. yap1Î? 2h 100uM AsIII, cad1Î? vs. cad1Î? 2h 100uMAsIII, rpn4Î? vs. rpn4Î? 2h 100uM AsIII, arr1Î? vs. arr1Î? 2h 100uM AsIII, parent vs. parent with 2h 100uM AsIII). The untreated specific deletion strain was also compared to the untreated parent strain in order to identify differential expression of genes due to the knockout alone. Keywords: other
2010-06-05 | E-GEOD-1798 | biostudies-arrayexpress
Project description:The abf1-1 mutant and wild type cells were grown at 30 degrees C in YPAD until OD600 reached 1.0-1.2. The same volume of 42 degrees C preheated YPAD was added for each flask. After 45 min incubation at 36 degrees C, cells were harvested and RNA was isolated. Four repeats of microarray results are in this series.
2010-06-09 | E-GEOD-1492 | biostudies-arrayexpress
Project description:Sarcomas are a biologically complex group of tumors of mesenchymal origin. By using gene expression microarray analysis, we aimed to find clues into the cellular differentiation and oncogenic pathways active in these tumors as well as potential biomarkers and therapeutic targets. We examined 181 tumors representing 16 classes of human bone and soft tissue sarcomas on a 12,601-feature cDNA microarray. Remarkably, 2,766 probes differentially expressed across this sample set clearly delineated the various tumor classes. Several genes of potential biological and therapeutic interest were associated with each sarcoma type, including specific tyrosine kinases, transcription factors, and homeobox genes. We also identified subgroups of tumors within the liposarcomas, leiomyosarcomas, and malignant fibrous histiocytomas. We found significant gene ontology correlates for each tumor group and identified similarity to normal tissues by Gene Set Enrichment Analysis. Mutation analysis done on 275 tumor samples revealed that the high expression of epidermal growth factor receptor (EGFR) in certain tumors was not associated with gene mutations. Finally, to further the investigation of human sarcoma biology, we have created an online, publicly available, searchable database housing the data from the gene expression profiles of these tumors (http://watson.nhgri.nih.gov/sarcoma), allowing the user to interactively explore this data set in depth. Keywords = sarcoma Keywords = cDNA Keywords = tumor profiling Common reference, multiclass gene profiling
2010-06-09 | E-GEOD-2553 | biostudies-arrayexpress
Project description:Duplicate hybridizations with fluorochrome switching were performed following the first two fractions of total body irradiation for accumulated doses of 1.5 Gy and 3.0 Gy. Control samples for all hybridizations were from the same patient before the beginning of treatment.
2010-06-09 | E-GEOD-1366 | biostudies-arrayexpress
Project description:This series contains microarrays for ten types of mouse lymphomas, nude mouse purified resting B cells, and nude mouse purified B cells stimulated with LPS. Microarray chips containing 70-mer oligonucleotides representing approximately 6,800 genes were prepared by the Microarray Center of the National Institute of Allergy and Infectious Diseases, Bethesda, MD. Keywords = mouse Keywords = lymphoma Keywords = B cells
2004-11-30 | E-GEOD-1908 | biostudies-arrayexpress
Project description:We investigated the effects of PACAP treatment, and endogenous PACAP deficiency, on infarct volume, neurological function, and the cerebrocortical transcriptional response in a mouse model of stroke, middle cerebral artery occlusion (MCAO). PACAP-38 administered i.v. or i.c.v. one hour after MCAO significantly reduced infarct volume, and ameliorated functional motor deficits measured 24 hours later in wild-type mice. Infarct volumes and neurological deficits (walking faults) were both greater in PACAP-deficient than in wild-type mice, but treatment with PACAP reduced lesion volume and neurological deficits in PACAP-deficient mice to the same level of improvement as in wild-type mice. A 35,546-clone mouse cDNA microarray was used to investigate cortical transcriptional changes associated with cerebral ischemia in wild-type and PACAP-deficient mice, and with PACAP treatment after MCAO in wild-type mice. 229 known (named) transcripts were increased (228) or decreased (1) in abundance at least 50% following cerebral ischemia in wild-type mice. 49 transcripts were significantly up-regulated only at one hour post-MCAO (acute response transcripts), 142 were up-regulated only at 24 hours post-MCAO (delayed response transcripts) and 37 transcripts were up-regulated at both times (sustained response transcripts). More than 50% of these are transcripts not previously reported to be associated with brain ischemia responses. Many of the acutely regulated neurotrauma-associated transcripts, but a larger percentage of the delayed ones, require endogenous PACAP for up-regulation by ischemia, suggesting a more prominent role for PACAP in later response to injury than in the initial response, and consistent with a neuroprotective role for PACAP in late response to injury and neuroprotective efficacy when given post-MCAO. Putative injury effector transcripts regulated by PACAP include ãÃ-actin, midline 2, and metallothionein 1. Potential neuroprotective transcripts include several demonstrated to be PACAP-regulated in other contexts. Prominent among these were transcripts encoding the PACAP-regulated gene Ier3, and the neuropeptides enkephalin, substance P (tachykinin 1), and neurotensin. In the hybridization method employed, equal amounts (2-5ãÃg/15.5 ãÃl) of total RNA obtained from brain tissue of different experimental groups were separately labeled with two different fluorescent dyes (Cy3 or Cy5) and applied on the same NIMH Mouse 36K cDNA microarray chip containing 13217 distinct Entrez gene IDs (formerly known as LocusLink) and 15888 distinct unigene IDs, the remainders are incompletely annotated IMAGE clones. Probe preparation and hybridization were performed as follows. The total RNA (15-50 ãÃg in 15.5 ãÃl) was incubated with amine-modified random primer (2 ãÃg/ãÃl, 2 ãÃl) and RNase inhibitor (5 units/ãÃl, 1 ãÃl) at 70 äaC for 10 min. Primer-RNA solution was then incubated at 42 äaC for 2 hr with the reverse transcriptase mix containing 5X first-strand buffer, 50X aa-dUTP/dNTPs (25 mM dATP, 25 mM dGTP, 25 mM dCTP, 15 mM dTTP and 10 mM aminoallyl dUTP), 0.1 M DDT, and Superscript II reverse transcriptase. The cDNA were labeled with NHS-ester Cy3 or Cy5 dye in the presence of 1 M sodium bicarbonate. Array slides placed in a hybridization chamber (Corning, Corning, NY) were incubated at 42 äaC for 16-24 hr, and successively washed with 0.5X SSC, 0.01% SDS, and 0.06X SSC at room temperature for 10 min each. Arrays were scanned with a GenePix 4000A scanner (Axon, Foster City, CA), and the resulting images were analyzed using IPlab (Fairfax, VA) and a FileMaker Pro 5 (Santa Clara, CA) relational database designed by Zedong Chen, NHGRI. Following background subtraction and normalization, a calculated ratio of Cy3 to Cy5 signal intensities was used to define the relative increase or decrease of a particular transcript. Ratios were calculated only from those spots with a combined ratio quality above 0.3.
2010-07-01 | E-GEOD-5902 | biostudies-arrayexpress
Project description:Experimental studies confirmed that seleium induced a global change of gene expression in HuH7 cells. Several novally induced secretory growth factors may contribute to the autocrine mode of growth elicited by selenium treatment. Keywords: Treatment HuH7 were cultured in 0.1 μM sodium selenite-containing medium for 15 passages (SELT) or in serum-free medium for 30 h (SF30) or in the regular complete medium containing 10 % fetal calf serum (HUH7) for 30 h. The total RNA were reverse transcried and hybridized to Human 8K-1 (Egenomix Technology Corp., Taiwan). The hybridization time and temperature was 16 h and 62â??, respectively. SELT and HUH7 samples were labeled with Cy5 while SF30 sample was labeled with Cy3. Local background correction and interchip global normalization were accomplished by GenePix Pro 5.0 (Molecular Devices, Sunnyvale, CA ). Genes with differential up- or down-regulation by 1.5 fold with respect to the SF30 cells were obtained using GeneSpring software 7.0 (Silicon Genetics, Agilent Technologies).
2010-06-30 | E-GEOD-4593 | biostudies-arrayexpress
Project description:Changes of genome-wide mRNA transcription levels of human ciliary smooth muscle (hCSM) cells were determined by treating hCSM cells in culture with 200 nM of either an prostaglandin E2 receptor subtype EP2 or subtype EP4 selective agonist for 6 hours in comparison to untreated controls. This was followed by competitive hybridization of fluorescent Cy3 or Cy5 labeled cRNA probes derived from the treated versus untreated control total RNA samples onto an Agilent Human Whole Genome Expression oligonucleotide microarray. Log 2 (LN) of the intra-slide ratios (RATIO, PRE_VALUE) of treated versus untreated samples was reported as VALUE in the sample files. Experiment Overall Design: Human Ciliary Smooth Muscle Cell Culture: Experiment Overall Design: Human ciliary smooth muscle (hCSM) cells were isolated from a female donor, received from the San Diego Eye Bank® (San Diego, CA) and cultured in DMEM with 10% fetal bovine serum and 0.5% penicillin/streptomycin according to the method reported previously by Woldemussie et al. Experiment Overall Design: For transcriptome analysis, cells were used for experiments starting at passage 4. Three independent experiments at consecutive cell passages were performed. For each experiment 0.6 x 106 cells were plated onto 15 cm dishes in MEM D-Valine medium (PromoCell, Germany) supplemented with 10% FCS, L-glutamine and antibiotics/antimycotics. Cells were grown to ~80% confluency for 6 days, followed by starvation in absence of FCS and presence of 100 nM of the cyclooxygenase inhibitor indomethacin, 20 µg/ml fatty acid-free BSA and 4 µg/ml transferrin for 17 hours. Cells were then exposed for 6 hours to 200 nM of either the EP2 agonist AGN210937 or the EP4 agonist AGN202280 (Allergan, Irvine, CA) or the corresponding amount of DMSO in the absence of FCS and presence of transferrin. Experiment Overall Design: Total RNA Extraction: Experiment Overall Design: Total RNA was isolated using a RNA isolation kit (RNAqueous; Ambion, USA) which was followed by DNase I treatment (Turbo DNA-free; Ambion) and purification, according to the manufacturerâ??s protocols. RNA was quantitated using a spectrophotometer ND-1000 (Nanodrop Technologies, USA). RNA quality was assessed regarding purity and stability using a Bioanalyser 2100 (Agilent Technologies, USA). Extracted total RNA aliquots were snap-frozen in liquid nitrogen and stored at -800C for single use. Experiment Overall Design: cRNA Labelling and Oligonucleotide Microarray Hybridization: Experiment Overall Design: Total RNA was linearly amplified and labelled with Cy3 or Cy5 using a low RNA input Fluor Linear Amp Kit (Agilent Technologies, USA). Internal RNA controls from Agilentâ??s RNA Spike-in Kit were included, comprised of mixtures of 11 in vitro synthesized transcripts derived from the Adenovirus E1A transcriptome at various ratios. The final cRNA concentration of 500 ng total RNA used was typically 590-650 ng/μl and the Cy3- or Cy5-cytidine incorporation was 5.4 -6.0 pmol/μg cRNA as determined using the Nanodrop spectrophotometer ND-1000. Experiment Overall Design: 3.5 μg of Cy3 and Cy5 labeled cRNA (treated vs. untreated samples) were competitively hybridized to high-density DNA arrays from Agilent Technologies (Whole Human Genome Oligo Microarray 44K) for 17 hours at 650C according to the manufacturerâ??s protocol. Each set of three microarray experiments for triplicate analysis contained one dye swap experiment. All microarrays were scanned and the intensities normalized over background as well as to eliminate signal intensity-dependent bias from the ratio of the two channels (Lowess normalization) using a microarray scanner from Agilent Technologies including proprietary software. All microarray data sets were imported into the microarray data analysis software Genespring 7.3 (Agilent Technologies) followed by comparison of normalized intensities. Gene identities were updated according to Agilentâ??s latest list of gene annotations (G4112F, 07Feb07). Experiment Overall Design: Consistency of microarray data quality was assessed by monitoring performance of the spiked internal RNA controls (330 microarray spots/array) at various ratios (Supplementary Data, Fig. S1), by carrying out a global analysis of all absolute fluorescent intensity values, and by analysis of negative control spots (314 microarray spots/array) for determination of detection thresholds.
2008-04-07 | E-GEOD-10621 | biostudies-arrayexpress
Project description:Human HaCaT keratinocytes were cultured to confluence (day10). Differentiated confluent cells were irradiated at 0.5 Gy or 2 Gy. RNA from irradiated cells was compared to RNA from non-irradiated reference cells in a dye-swap hybridization procedure 24 h after irradiation. Keywords = human keratinocytes, gamma irrdiation
2003-03-25 | E-GEOD-356 | biostudies-arrayexpress