Transcriptomics of HL60 cell line at 30 min and 1 h after all-trans-retinoic acid (ATRA) treatment
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ABSTRACT: Transcriptome profiling of HL60 cells at the yearly time points of ATRA treatment (30 min and 1 h) allows to unveil molecular mechanisms of the onset of granulocytic maturation. Transcriptomic techniques are characterized by high sensitivity and allow for detection of low-abundant molecules, such as nuclear transcription factors.
Project description:Transcriptional profiling of HL-60 cells at 0, 3, 24, and 96 h after ATRA treatment. The differentially expressed genes was meant to be the input data for bioinformatic upstream regulator search
Project description:The nuclear proteome harbors a sheer number of regulatory proteins, including transcription factors (TFs). Profiling of nuclear proteome during all-trans-retinoid acid(ATRA)-induced differentiation of HL60 cells allows to unveil molecular mechanisms of granulocytic maturation. It is especially important to have an understanding of molecular perturbations at the early stages of the differentiation process. Applying proteomic profiling using isobaric labeling coupled with alkaline fractionation (TMT/2D) we identified 1860 nuclear proteins with high confidence (FDR<0.01, at least 2 unique peptides per protein). Among them 136, 226, 280, 312 and 241 proteins were found to be altered at 3, 6, 9, 12, and 72 h in HL60 cell nuclear fraction under ATRA treatment.
Project description:Gene expression profiles of in vitro selected highly metastatic MKN45-GFP sublines. The results were compared with MKN45-GFP control cell line to determine the metastasis associated genes. Four pairs compared experiment. Each pair was used MKN45-GFP cells as correlated control. Determining on the gene expression trends were by various metastatic ability of each subline.
Project description:Damage of the intestinal epithelial barrier by xenobiotics or reactive oxygen species and a dysregulated immune response are both factors involved in the pathogenesis of inflammatory bowel diseases (IBD). Curcumin and rutin are polyphenolic compounds known to have anti-oxidant and anti-inflammatory activities, but their mechanism(s) of action are yet to be fully elucidated. Mdr1a-/- mice spontaneously develop intestinal inflammation, predominantly in the colon, with pathology similar to IBD, so this mouse model is relevant for studying diet-gene interactions and potential effects of foods on remission or development of IBD. This study tested whether the addition of curcumin or rutin to the diet would alleviate colonic inflammation in mdr1a-/- mice. Using whole-genome microarrays, the effect of dietary curcumin on gene expression in colon tissue was also investigated. Twelve mice were randomly assigned to each of three diets; control (AIN-76A), control + 0.2% curcumin or control + 0.1% rutin and monitored from the age of 7 to 24 weeks. Curcumin, but not rutin, significantly reduced histological signs of colonic inflammation in mdr1a-/- mice. Microarray and pathway analyses suggested that the effect of dietary curcumin on colon inflammation could be via an up-regulation of xenobiotic metabolism and a down-regulation of pro-inflammatory pathways probably mediated by PXR and PPARï¡ activation of RXR. These results reveal the potential of global gene expression and pathway analyses to study and better understand the effect of foods in colonic inflammation. Experiment Overall Design: Twelve mice were randomly assigned to each of three diets; control (AIN-76A), control + 0â¢2% curcumin or control + 0â¢1% rutin and monitored from the age of 7 to 24 weeks. As only curcumin significantly reduced colonic HIS, comparison of the gene expression levels in colon was carried out using total RNA from colon tissue of four mdr1a-/- mice from the control group (high HIS) and four mdr1a-/- mice from the curcumin group (low HIS). A reference design with eight arrays was used for this comparison, where each individual RNA sample was hybridized in the array with the reference RNA, totalizing 4 biological replicates per treatment.
Project description:Municipal wastewater effluent can impact its receiving environment. In the St. Lawrence River, male fish living downstream from Montreal exhibit increased hepatic vitellogenin, intersex, delayed spermatogenesis and altered immune function. Few studies have examined genome-wide effects associated with municipal effluent exposure in fish to decipher the mechanisms of toxicity. The present objective was to identify hepatic cellular signaling pathways in fathead minnows following exposure to municipal wastewater effluent. Immature minnows were exposed for 21 days to either 0% (Control) or 20% municipal effluent, the highest concentration in the St. Lawrence River. Hepatic RNA was extracted and used to hybridize a fathead minnow oligonucleotide microarray containing approximately 15K gene sequences. Sixteen samples were examined, 8 control samples and 8 exposed samples.
Project description:Plasma-derived extracellular vesicles (EVs) have been extensively described as putative biomarkers in different diseases. More precisely, increased levels of EVs subpopulations are well-known to associate with obesity. The goal of this study was to identify EVs-derived biomarkers in plasma from obese patients in order to predict the development of pathological events associated with obesity. The study was performed combining two different proteomic approaches (2D-DIGE and label-free LC-MS/MS). Samples were obtained from 22 obese patients, with no associated comorbidities, and their lean-matched controls. EVs were isolated following a serial ultracentrifugation protocol. We detected 22 and 23 different regulated protein features from 2D-DIGE and label free LC-MS/MS respectively. Most of the differentially regulated proteins identified were involved in the coagulation and complement cascade. Interestingly, there was a clear up-regulation of complement C3, complement C4 and fibrinogen in obese patients following both approaches, which correlates with a pro-inflammatory and prothrombotic state of those individuals. On the other hand, we showed a down-regulation of adiponectin leading to an increased risk of suffering cardiovascular diseases. Our results suggest the relevance of considering plasma-derived-EVs proteins as a source of potential biomarkers for the development of atherothrombotic events in obesity.
Project description:The objective of our study was to search for survival biomarkers (SB) and treatment response monitoring biomarkers (TRMB) in the urinary proteome of dogs with renal disease secondardy to canine leishmaniosis (CanL),
Project description:Gene expression profile comparing 17-month old wild-type with psen1hu2547 zebrafish. Total RNA from 3-4 brains of 17-month old adult psen1hu2547 or WT fish was extracted using RNeasy mini columns (Qiagen) and quality-checked using NanoDrop 1000 (Thermo Scientific) and formaldehyde gel electrophoresis. cDNA was generated and labeled with Cy3 or Cy5-CTP using the low RNA input linear amplification kit (Perkin-Elmer). Upon purification and fragmentation, labeled cRNA was hybridized to the zebrafish 22K 60-mer oligonucleotide microarray (G2518A-001, Agilent Technologies, Santa Clara, CA, USA) at 65ºC for 17 hours. Three independent biological repeats were performed.
Project description:In this study, we performed temporal profiling of transcriptome and chromatin accessibility in HL-1 cells for understanding the molecular mechanisms underlying cardiac responses to hypoxia. We collected HL-1 cells under four conditions (4 h and 8 h of hypoxia exposure, 24 h reoxygenation and the normal condition), applied RNA-seq and ATAC-seq to them and performed pairwise comparison of gene expression and open chromatin status on a genome-wide scale.