Transcriptome-wide analysis of mRNA decay in normal and osteoarthritic human articular chondrocytes
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
SUBMITTER: Simon Tew
PROVIDER: E-MTAB-2723 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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2016-05-17 | E-GEOD-78209 | biostudies-arrayexpress
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2011-07-29 | E-GEOD-26829 | biostudies-arrayexpress
Project description:We performed whole-genome stability measurements for MDA-MB-231 and its highly metastatic derivative MDA-LM2. Our goal was to identify post-transcriptonal regulons that are deregulated en route to higher metastatic capacity. Cells were pulsed with 4-thiouridine for 2 hours and then RNA was extracted at 0, 2, 4, and 7 hr time-points in quadruplicate from each cell line. 4sU labeling followed by RNA-seq was then used to measure the abundance of transcripts in each population. A decay rate was estimated based on the rate at which transcript abundance was reduced at these time-points.
2014-07-24 | E-GEOD-49608 | biostudies-arrayexpress
Project description:Total protein extracts (days 1, 2, 3 and 4 of the growth curve; separation of peanut lectin-agglutinating and non-agglutinating choanomastigotes at day 4). Taxonomy: Crithidia fasciculata (Kinetoplastida, Trypanosomatidae)
2015-07-10 | PXD001331 | Pride
Project description:Wild-type and talA talB double knockout cells were grown in 50 ml of MOPS medium supplemented with 0.2% glucose or xylose in 500 ml Erlenmeyer flasks until the OD600 reached 1. At the sampling RNA was stabilized by mixing with RNAprotect Bacteria regent (Qiagen) and total RNA were isolated, by RNeasy mini Kit (Qiagen). Duplicate experiments employing dye swapping were performed for comparison. Protocol and conditions used for hybridization and washing were as described {Oshima, 2002 #712}. Images were scanned by Affymetrix 428 in 10µm resolution using Jaguar 2.0 software and processed by Imagene 4.0 software (BioDiscovery, Segundo ,USA) by default setting.?The raw numerical data files from Imagene were further processed by GeneSpring 7.3 (Agilent) software package. A Lowess curve was fit to the log-intensity versus log-ratio plot. 20.0% of the data was used to calculate the Lowess fit at each point. This curve was used to adjust the control value for each measurement.
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2014-11-17 | E-MTAB-2123 | biostudies-arrayexpress
Project description:Certain oncolytic viruses exploit activated Ras signalling in order to replicate in cancer cells. Constitutive activation of the Ras/MEK pathway is known to suppress the effectiveness of the interferon (IFN) antiviral response, which may contribute to Ras-dependent viral oncolysis. Here, we identified 10 human cancer cell lines (out of 16) with increased sensitivity to the anti-viral effects of IFN-α after treatment with the MEK inhibitor U0126, suggesting that the Ras/MEK pathway underlies their reduced sensitivity to IFN. To determine how Ras/MEK suppresses the IFN response in these cells, we used DNA microarrays to compare IFN-induced transcription in IFN-sensitive SKOV3 cells, moderately resistant HT1080 cells, and HT1080 cells treated with U0126. We found that 267 genes were induced by IFN in SKOV3 cells, while only 98 genes were induced in HT1080 cells at the same time point. Furthermore, the expression of a distinct subset of IFN inducible genes, that included RIGI, GBP2, IFIT2, BTN3A3, MAP2, MMP7 and STAT2, was restored or increased in HT1080 cells when the cells were co-treated with U0126 and IFN. Bioinformatic analysis of the biological processes represented by these genes revealed increased representation of genes involved in the anti-viral response, regulation of apoptosis, cell differentiation and metabolism. Furthermore, introduction of constitutively active Ras into IFN sensitive SKOV3 cells reduced their IFN sensitivity and ability to activate IFN-induced transcription. This work demonstrates for the first time that activated Ras/MEK in human cancer cells induces downregulation of a specific subset of IFN-inducible genes. HT1080 cancer cells treated for 6 hours or 12 hours with interferon-alpha (500U/ml), the MEK inhibitor U0126 (20uM) or both, triplicate biological samples (18 samples). SKOV3 cells treated with interferon-alpha (500U/ml) for 6h, triplicate biological samples (6 samples).
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Project description:Background: We seek to understand correlates of chronic stress and their influence on the development of substance use in a population-based cohort of young adults (OYSUP). Methodology: We evaluated clinical features, salivary RNA metrics and differential expression of candidate genes in unfractionated saliva samples from 48 individuals (31% female, 55% ever-smoking) randomly selected from two groups stratified by high versus low severe negative life events and difficulties within the last year as as measured by the Life Events and Difficulties Schedule (LEDS, Brown & Harris, 1978). We used multiple analytical platforms, multiple reference genes and 18 replicates of all gene expression assays (assays). We chose 38 candidate genes previously identified as differentially expressed in a genome-wide scan of monocyte RNA in 11 familial caregivers of glioblastoma multiforme patients and 10 control subjects matched on age (middle-aged), gender (57% female), ethnicity and marital status and free of major stressors during the previous year. Principal Findings: We observed significant differences in smoking prevalence (Pearson’s ?22d.f.=4.5, P=0.035) and RNA integrity score (t=2.12, P=0.039) between stress strata. We analyzed quantitative PCR data using both the comparative standard curve (CSC) and relative standard curve (RSC) methods because 34% of assays had efficiency estimates <90%. Assay data exhibited excellent linearity and low variability. We observed significant under expression of five assays in the high stress stratum using both methods, four of which interrogate glucocorticoid receptor regulated genes. Post-hoc analyses identified significant associations of RNA yield, integrity score, gender, ever-smoking and/or stress with results of normalized gene expression in 22/37 assays. IL8 expression remained significantly associated with chronic stress after adjustment for clinical and salivary RNA variables. Salivary IL8 expression has been previously associated with oropharyngeal squamous cell cancer. Conclusions: A gene expression signature of chronic stress previously observed in adult hematopoietic samples is observed in the unfractionated saliva of young adults. Future investigation will need to analyze associated clinical, metabolic, meta-genomic, and proteomic components of unfractionated saliva to elucidate factors influencing the salivary transcriptome. Gene expression was performed on a Fluidigm 48 x 48 array on 48 individuals for 38 target genes and 6 possible reference genes (represented by 9 reference assays) in triplicate. Quantitative Q-PCR datas were normalized according to relative levels of four reference genes (CDKN1B, YWHAZ, RPL13A, and UBC).
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