Project description:DNA methylation profiling of gene promoters in the human myeloid cell line U937. Bisulphite converted DNA from wildtype U937 cells was hybridised to the Illumina Infinium HumanMethylation27 BeadChip to determine the methylation pattern of the U937 cell line.
Project description:DNA methyltransferase 3A (DNMT3A) gene is mutated in various myeloid neoplasms including acute myeloid leukemia (AML), especially at the Arg882 and associated with inferior outcomes. Despite the current progress of functional role of DNMT3A mutations, the molecular pathogenesis of myeloid malignancies remains poorly understood. The mechanisms of AML transformation and functional role of DNMT3A mutations through its target genes in the leukemogenesis remain to be explored. Here we wished to perform the differential genomic-methylation profile in U937 cells over-expressed with DNMT3A-Arg882His/Cys (R882H/C) mutations including DNMT3A-WT and vector using Illumina MethylationEPIC BeadChip microarray. Results: Differential genomic-methylation assess identified both hypo- and hypermethylation features in different regions throughout the whole genome of DNMT3A mutants-transduced U937 cells.
Project description:This study aims to determine the chromosome content and organisation of two myeloid leukaemia cell lines, HEL and U937. This will be done not only with SNP array data to determine breakpoints and copy number of copy number aberrations, but also with FISH (multicolour FISH, multicolour banding, centromere and single locus FISH) to identify translocation partners, chromosome organistion, centromere content, and provide some information on genome evolution in the cell line. Although several HEL SNP array karyotypes have been published and are available online the information here shows that there are some differences, and the additional FISH tests provide a greater depth of information on genome organisation and derivation of the abnormal chromosomes. The U937 cell line was also studied using DNA from fresh and fixed specimens for comparison of the quality of the SNP array data. Data from cells fixed using standard cytogenetic fixative (3:1 methanol:acetic acid) were compared to data from cells processed directly from tissue culture.
Project description:Pirin (PIR) is a putative transcriptional regulator whose expression is silenced in cells bearing the AML1/ETO and PML/RAR leukemogenic fusion proteins and is significantly repressed in a large proportion of acute myeloid leukemias. PIR expression increases during in vitro myeloid differentiation of primary hematopoietic precursor cells, and ablation of PIR in the U937 myelomonocytic cell line or in murine primary hematopoietic precursor cells results in impairment of terminal myeloid differentiation. Keywords: Transcriptional regulation, knock-down using shRNA We analyzed gene expression profiles of U937 cells after knock-down of PIR using the shPIR#7 oligonucleotides cloned in the pSICO-R lentiviral vector. No residual PIR protein is detectable in U937-shPIR#7 cells by Western blotting. U937 cells containing the empty cloning vector (U937-pSICO) were used as control. shPIR#7 oligonucleotide sequences: Human PIR#7-for: TGAAGCCACTTTGTCTTAATTTCAAGAGAATTAAGACAAAGTGGCTTCTTTTTTC Human PIR#7-rev: TCGAGAAAAAAGAAGCCACTTTGTCTTAATTCTCTTGAAATTAAGACAAAGTGGCTTCA For each sample, an RNA pool was obtained by mixing equal quantities of total RNA from each of three independent RNA extractions. Each biotin-labeled target was hybridized to two GeneChip HG-U133 Plus v.2 arrays.
Project description:We analyzed the transcriptional effect of IRF2 in the U937 human monocyte cell line. The major findings of the study is the regulation of CASP4 by IRF2
Project description:This SuperSeries is composed of the following subset Series: GSE10520: Genes regulated by AML1/ETO in U937 cells GSE10537: Gene expression profiling of AML1/ETO regulated genes and binding pattern on human promoters in U937 cells GSE10578: AML1/ETO, AML1, and HEB binding patterns on chromosme 19 GSE10579: Analysis of expression levels of genes on chromosome 19 in U937 cells expressing AML1/ETO Keywords: SuperSeries Refer to individual Series
Project description:We employed the AML cell line model U937 to determine the effects of single and dual treatment with the HMA decitabine and the HDACi Pano or VPA on gene expression and DNA methylation. We noted global alterations of gene expression and a massive effect on the methylome.
Project description:Pirin (PIR) is a putative transcriptional regulator whose expression is silenced in cells bearing the AML1/ETO and PML/RAR leukemogenic fusion proteins and is significantly repressed in a large proportion of acute myeloid leukemias. PIR expression increases during in vitro myeloid differentiation of primary hematopoietic precursor cells, and ablation of PIR in the U937 myelomonocytic cell line or in murine primary hematopoietic precursor cells results in impairment of terminal myeloid differentiation. Keywords: Transcriptional regulation, knock-down using shRNA