Induced expression of the C19MC miRNAs in HTR-8/SVneo cells (mRNA)
Ontology highlight
ABSTRACT: Analyze gene expression profiles in HTR-8/SVeo cells stably transfected with a BAC plasmid containing the entire C19MC miRNA cluster HTR-8/SVeo cells were transfected with a modified BAC plasmid containing the entire C19MC miRNA cluster and carrying a zeocin selection cassette. Several independent clones and a mixed population of transfected cells were analyzed and compared to non-transfected HTR-8/SVeo cells and primary human trophoblasts that express the C19MC miRNAs endogenously
Project description:Analyze miRNA expression levels in HTR-8/SVeo cells stably transfected with a BAC plasmid containing the entire C19MC miRNA cluster HTR-8/SVeo cells were transfected with a modified BAC plasmid containing the entire C19MC miRNA cluster and carrying a zeocin selection cassette. Several independent clones and a mixed population of transfected cells were analyzed and compared to non-transfected HTR-8/SVeo cells and primary human trophoblasts that express the C19MC miRNAs endogenously
Project description:Analyze gene expression profiles in HTR-8/SVeo cells stably transfected with a BAC plasmid containing the entire C19MC miRNA cluster
Project description:Analyze miRNA expression levels in HTR-8/SVeo cells stably transfected with a BAC plasmid containing the entire C19MC miRNA cluster
Project description:SPARC is a matricellular glycoprotein that plays critical roles in the pathologies associated with obesity and diabetes, as well as tumorigenesis. The objective of this study was to investigate the role of SPARC in the process of trophoblast invasion which shares many similarities with tumor cells invasion. Our results reveals that hormones, cell adhesion molecules, ECM molecules, growth factors and cytokines all are mediated by SPARC in EVT invasion. HTR-8/SVneo cells were transfected with SPARC siRNA or a 25-nucleotide universal negative control siRNA using Lipofectamine 2000 according to the manufacturer’s protocol. Seventy-two hours after transfection, cells were harvested and RNA was isolated using standard procedures.
Project description:IL-11 increases the invasiveness of JEG-3 cells while, reduces the invasiveness of HTR-8/SVneo cells. This study examines the effect of IL-11 on gene expression in trophobalstic cell models viz. JEG-3 and HTR-8/SVneo cells to resolve the controversies associated with the IL-11 mediated regulation of the invasiveness of these two cell lines. JEG-3 and HTR-8/Svneo cells were stimulated with IL-11 (200 ng/ml) for 24 h in pain medium keeping un-treated cells as control. After 24 h of stimulation, total RNA was isolated from these cells and used for the microarray experiments. These experiments were performed once.
Project description:Both leukemia inhibitory factor (LIF) and interleukin-6 (IL-6) increase the invasiveness of JEG-3 and HTR-8/SVneo cells. This study examines the effect of LIF and IL-6 on gene expression in trophoblastic cell models viz. JEG-3 and HTR-8/SVneo cells to decipher the molecular basis of the increase in invasiveness. JEG-3 and HTR-8/Svneo cells were stimulated with LIF (50 ng/ml) and IL-6 (100 ng/ml) for 24 h in plain medium keeping untreated cells as a control. After 24 h of stimulation, total RNA was isolated from these cells and used for microarray experiments. These experiments were performed once.
Project description:Abnormal trophoblast invasion is associated with the most common and most severe complications of human pregnancy. The biology of invasion, as well as the etiology of abnormal invasion remains poorly understood. The aim of this study was to characterize the transcriptome of the HTR-8/SVneo human cytotrophoblast cell line which displays well characterized invasive and non-invasive behaviors, and to correlate the activity of the transcriptome with nuclear matrix attachment and cell phenotype. Interestingly comparison of the transcriptome did not reveal an obvious significant difference between the transcriptomes of invasive and non-invasive HTR cells. In contrast, comparison of the MARs on chromosomes 14-18 revealed an increased number of MARs associated with an invasive phenotype. These attachment areas were more likely to be associated with silent (rather than actively transcribed) genes. DNA extraction with a 2 M NaCl solution followed by restriction digestion with EcoR1 facilitates the separation of matrix bound DNA from non matrix bound DNA. Genomic CGH arrays were used to map the relative differences between attached (scaffold/matrix) and non-attached (loop) portions of the genome in non-invasive (proliferative) and invasive trophoblasts. The matrix association was assessed relative to gene expression measured on Illumina expression beadchips.
Project description:Immature cell populations, including stem cells and progenitor cells, can be found in “side-population (SP)” cells. Although SP cells isolated from some adult tissues have been reported elsewhere, isolation and characterization of human trophoblast SP cells remained to be reported. We used microarrays to detail the global program of gene expression underlying cell differentiation and identified up-regulated genes of SP cells. HTR-8/SVneo SP cells or NSP cells were isolated using a cell sorter for RNA extraction and hybridization on Affymetrix microarrays. To obtain many SP or NSP cells, we cultured a large amount of HTR-8/SVneo cells (1×10^7 cells). The cells were labeled with 2.5µg/ml Hoechst 33342 dye (Molecular Probes, Eugene, OR) for 120 minutes at 37 ℃, either alone or in combination with 50 µmol/L verapamil (Sigma-Aldrich, ST Louis, MO). Finally, the cells were then passed through EPICS ALTRA HyPerSort (Beckman Coulter, Fullerton, CA) using dual wavelength analysis (blue, 424-444 nm; red, 675 nm) after excitation with 350 nm UV light.
Project description:Immature cell populations, including stem cells and progenitor cells, can be found in M-bM-^@M-^\side-population (SP)M-bM-^@M-^] cells. Although SP cells isolated from some adult tissues have been reported elsewhere, isolation and characterization of human trophoblast SP cells remained to be reported. We used microarrays to detail the global program of gene expression underlying cell differentiation and identified up-regulated genes of SP HTR-8/SVneo SP cells or NSP cells were isolated using a FACS Vantage fluorescence-activated cell sorter (BD Bioscience, San Jose, California) using dual wavelength analysis (blue, 424 - 444nm; red 675 nm) after excitation with 350 nm UV light. PI-positive dead cells were excluded from the analysis. Sorted SP and NSP cells' RNA had been analyzed by microarray analysis.