Project description:Tamoxifen is an anti-estrogen drug used in the treatment of Estrogen Receptor (ER) positive breast cancer. Effects and side effects of tamoxifen is the sum of tamoxifen and all its metabolites. Using concentrations that mimic the clinical situation we examined effects of 4OHtam, 4OHNDtam and NDtam on global gene expression in 17beta-estradiol (E2) treated MCF-7 cells.
Project description:Determine differentially expressed human genes in A549 tumors when co-cultured in spheroids with mouse fibroblast harbouring a mutation in the heparan sulphate elongation enzyme Ext1 compared to corresponding wild-type fibroblast/A549-containing spheroids.
Project description:The aim was to compare the global gene expression of epididymal white adipose tissue (eWAT) of WT and Irx5-KO mice. Mice 10 weeks of age were fed a high-fat diet for 10 weeks before eWAT was dissected out, RNA extracted and microarray performed
Project description:The aim of the project was to compare global gene expression in adipocytes from obese patients and lean controls. Subcutaneous adipose tissue was collected from severely obese patients undergoing bariatric surgery (average body-mass index (BMI) of 45.5 kg/m2 (n = 12, thereof 4 men) and healthy lean patients undergoing hernia repairs (average BMI of 24.2 kg/m2 (n = 12, thereof 7 men), between 27 and 56 years of age. Adipocytes were isolated by collagenase treatment of adipose tissue, followed by filtering and centrifugation. Floating adipocytes were lysed in Qiazol before RNA purification and microarray analysis.
Project description:We generated 77-bp Illumina reads from single messenger RNA libraries from four diverse developmental stages of the two-spotted spider mite to maximally capture the complement of transcribed sequences across development. Adult, nymphal, larvae and embryonic stages were separated using sieves of various pore sizes, and mites of various developmental stages were carefully selected for transcriptome library preparation. Samples were a mix of males and females to capture male and female patterns of transcription, and were reared on beans (Phaseolus vulgaris cv California Red Kidney). The RNA-Seq data was used for validation of gene models predicted by EuGene, and to study patterns of gene expression across development. Gene expression for spider mites from adult, nymph, larvae and embryonic developmental stages was examined (technical replicates were generated).
Project description:Pseudogenes are thought to be inactive gene sequences, but recent evidence of extensive pseudogene transcription raised the question of potential function. Here we discover and characterize the sets of lncRNAs induced by inflammatory signaling via TNFα. TNFα regulates hundreds of lncRNAs, including 54 pseudogene lncRNAs, several of which show exquisitely selective expression in response to specific cytokines and microbial components in a NF-κB-dependent manner. Lethe, a pseudogene lncRNA, is selectively induced by proinflammatory cytokines via NF-κB or glucocorticoid receptor agonist, and functions in negative feedback signaling to NF-κB. Lethe interacts with NF-κB subunit RelA to inhibit RelA DNA binding and target gene activation. Lethe level decreases with organismal age, a physiological state associated with increased NF-κB activity. These findings suggest that expression of pseudogenes lncRNAs are actively regulated and constitute functional regulators of inflammatory signaling. RNA profiles of wild type (WT) MEFs treated with TNF-alpha were generated by deep sequencing using Illumina GAIIx. Examination of H3K4me3 histome modification in MEF.
Project description:Tumor-associated stromal cells can enable cancer cells to become insensitive to therapy. They can promote aggressive phenotype in cancer cells, which become less responsive to drugs such as BRAF inhibitors (BRAFi) used to treat melanomas. To clarify potential mechanism behind stromal influence on melanoma, we analyzed gene expression in Melmet 5 melanoma cells grown as mono-cultures or co-cultures with lung fibroblasts with/without BRAFi. We have shown that Melmet 5 growing as co-cultures gained a de-differentiated, invasive transcriptional state, which is known to be linked to BRAFi-resistance. The transcriptional changes induced by BRAFi were much larger in Melmet 5 mono-cultures compared to co-cultures, indicating a much dampened transcriptional response to BRAFi in melanoma under the influence of fibroblasts. We conclude that interaction with the stromal cells stimulate melanoma cell transition to the invasive de-differentiated phenotype, leading to a worse response to BRAF inhibitors. Total RNA was isolated from Melmet 5 cell line growing as mono- or co-cultures with fibroblasts for 72 hours and treated with BRAFi for the last 24 hours.
Project description:In order to elucidate the role of DNA methylation in the DME gene regulation, global DNA methylation and mRNA expression profiles of human tissues and cell lines were examinde by HumanMethylation450 Bead Chip and SurePrint G3 Human Gene Expression 8×60K v2. We demonstrated DNA methylation landscape of the DME genes in human tissues. Although a fraction of DME genes can be regulated by their DNA methylation, the variable DNA methylation status probably affects drug metabolism and response. Bisulphite converted DNA from the 4 samples were hybridized to the Illumina HumanMethylation450 BeadChip (Sample L and SI were examined twice).
Project description:The integrin α11-overexpressing transgenic mouse strain was generated to further understand the function of α11, after generation of our integrin α11-KO strain. Analysis of the strain showed that integrin α11 was highly expressed in heart, which correlated with a fibrotic heart phenotype. To understand the molecular mechanisms underlying this phenotype, we then performed Microarray to investigate the differentially expressed genes between the WT and TG mice (α11-overexpressing mice) which may result from integrin α11 overexpression. The microarray was performed on RNA-isolates from the heart apex of 1-year-old WT (n=4) and α11-TG (n=6) mice. Total RNA isolation was performed using RNeasy Fibrous Tissue Mini Kit (Qiagen) in accordance with the manufacturerâs protocol. Quality of RNA samples was assessed using Agilent Bioanalyzer 2100. Microarray was carried out at the Norwegian Microarray Consortium/University of Bergen Microarray Core Facility at Haukeland University Hospital. Illumina Bead Array Technology was used, and the raw microarray data was quality examined in GenomeStudio. Differentially expressed genes were analysed using the Ingenuity Pathways Analysis (IPA) software (Ingenuity Systems, Redwood City, CA).
Project description:Pseudogenes are gene copies presumed to mainly be functionless relics of evolution due to acquired deleterious mutations or transcriptional silencing. When transcribed, pseudogenes may encode proteins or enact RNA-intrinsic regulatory mechanisms. However, the extent, characteristics and functional relevance of the human pseudogene transcriptome are unclear. Short-read sequencing platforms have limited power to resolve and accurately quantify pseudogene transcripts owing to the high sequence similarity of pseudogenes and their parent genes. Using deep full-length PacBio cDNA sequencing of normal human tissues and cancer cell lines, we identify here hundreds of novel transcribed pseudogenes. Pseudogene transcripts are expressed in tissue-specific patterns, exhibit complex splicing patterns and contribute to the coding sequences of known genes. We survey pseudogene transcripts encoding intact open reading frames (ORFs), representing potential unannotated protein-coding genes, and demonstrate their efficient translation in cultured cells. To assess the impact of noncoding pseudogenes on the cellular transcriptome, we delete the nucleus-enriched pseudogene PDCL3P4 transcript from HAP1 cells and observe hundreds of perturbed genes. This study highlights pseudogenes as a complex and dynamic component of the transcriptional landscape underpinning human biology and disease.