Gene based expression changes in glioblastoma cells after downregulation of MPS1 kinase
ABSTRACT: Gene expression changes were analyzed in U251 GBM cells after downregulation of MPS1 by RNA interference technology at different time points Microarray analysis was used to compare the mRNA expression profile of siMPS1 silenced U251 cells compared to siNegative (siNeg) and untransfected (Control) cells at 6, 24 and 48 hours post tranfection
UNLABELLED:To ensure faithful chromosome segregation, cells use the spindle assembly checkpoint (SAC), which can be activated in aneuploid cancer cells. Targeting the components of SAC machinery required for the growth of aneuploid cells may offer a cancer cell-specific therapeutic approach. In this study, the effects of inhibiting Monopolar spindle 1, MPS1 (TTK), an essential SAC kinase, on the radiosensitization of glioblastoma (GBM) cells were analyzed. Clonogenic survival was used to determi ...[more]
Project description:Gene expression changes were analyzed in U251 GBM cells after downregulation of MPS1 by RNA interference technology at different time points Overall design: Microarray analysis was used to compare the mRNA expression profile of siMPS1 silenced U251 cells compared to siNegative (siNeg) and untransfected (Control) cells at 6, 24 and 48 hours post tranfection
Project description:U251 gliolbastoma cells were treated with vehicle (DMSO) or G-TPP, a mitochondrial targeted version of 17-AAG. 6 hours after treatment RNA was harvested and subjected to microarray analysis on Human Gene 1.0 ST Array. We sought to determine a mitochdondrial gene response after targeting inhibition of mitochondrial chaperones, Hsp90.
Project description:Gene expression profiling of SNB19 and U251 glioblastoma cell lines transfected with the FGFR3-TACC3 fusion, FGFR3 wildtype and TACC3 wildtype constructs. SNB19 and U251 cells were transfected with different clones of the FGFR3-TACC3 fusion and with wildtype FGFR3 and TACC3 constructs. Total RNA was extracted and hybridized onto Agilent dual channel gene expression microarrays. In each hybridization, empty vector transfected SNB19 or U251 cells were hybridized into the reference channel.
Project description:WT-1 is a zinc-finger transcription factor with important roles during development. WT-1 is associated with the development of tumors of varied origins. Our lab has previously demonstrated aberrant expression of WT-1 in tumor cells. Necessity of WT-1 for tumor cell growth- in vitro & in vivo- was also shown. Human glioma U251 cells were treated with Oligofectamine (CTRL), Oligofectamine + non-targeting siRNA control (SCR) or Oligofectamine+ si-WT1 siRNA (Si). Biological triplicates were run on microarrays.
Project description:Enterovirus 71 (EV71) is one of the leading causes of hand, foot and mouth disease with neurological complications in some cases. To study the pathogenesis of EV71 infection, large scale analyses of EV71 infected cells have been performed. However, most of these studies employed rhabdomyosarcoma (RD) cells or used transcriptomic strategy. Here, we performed SILAC-based quantitative proteomic analysis of EV71-infected U251 cells, a human glioma cell line. A total of 3,125 host proteins were quantified, 451 of which were differentially regulated as a result of EV71 infection at 8 hpi or 20 hpi or both.
Project description:We identified GSK3 as a regulator of GBM cell survival using microarray analysis, small molecule and genetic inhibitors of GSK3 activity. Various molecular and genetic approaches were then employed to dissect out the molecular mechanisms responsible for GSK3 inhibition-induced cytotoxicity. Experiment Overall Design: RNA extracted from U251 cells treated with DMSO or Enzastaurin for indicated time were hybridized to Affymetrix expression arrays (HG_U133-Plus_2) to detect changes in gene expression caused by Enzastaurin. Three replicates per sample were used.
Project description:The conserved RNA-binding protein Musashi1 (MSI1) has emerged as a key oncogenic factor in numerous solid tumors, including glioblastoma. However, its mechanism of action has not yet been established comprehensively. We set out to map its impact on the transcriptome in U251 cells using RNA-seq and iCLIP. Examination of gene expression and splicing changes upon KD of Musashi1 in U251 cells and link to iCLIP-identified Musashi1 RNA binding sites
Project description:To evaluate gene expression alteration following miR-139-5p transfection in glioma cells. We find a significant downregulation of two transcriptional factors, E2F3 and HoxA9. Total RNA were extracted from U87, LN229 and U251 glioma cells transfected with miR-139-5p or miRNA negative control.
Project description:To identify the epigenetic signature that controls cancer cell migration, we performed integrated gene expression, miRNA and epigenetic profiling of 486 selected invasion-associated genes in non-migratory MCF-7 breast carcinoma and highly migratory U251 glioma cells. We used a custom-built chromatin immunoprecipitation (ChIP)-on-Chip microarray that included complimentary oligonucleotide probes for the genes that were functionally linked to proteolysis, migration and tumorigenesis such as extracellular matrix proteins, cellular proteinases and their inhibitors, growth factors and cytokines, and adhesion and signaling receptors. As a result, we determined the role histone H3 modifications [(Lys-4 dimethylation (H3K4me2), Lys-27 trimethylation (H3K27me3) and acetylation (H3ac)] play in transcriptional regulation of the multiple invasion-associated genes. Predominantly, transcriptional silencing of the pro-invasive genes in MCF-7 cells involved the repressive H3K27me3 mark and, frequently, the presence of the stem cell-like bivalent epigenetic mark (enrichment in both H3K27me3 and H3K4me2). In turn, pro-invasive genes in U251 cells were epigenetically stimulated by a gain in H3K4me2 and histone H3 hyperacetylation, and by a global reduction of H3K27me3. Intriguingly, the expression of multiple collagen genes was highly enhanced in glioma cells, thus suggesting that gliomas themselves deposit a specialized, invasion-promoting matrix. miRNA global profiling complements the ChIP-on-Chip experiment with U251 and MCF-7 cells. Two-condition experiment, MCF7 vs.U251 cells. 2 Biological replicates for each cell line
Project description:We have found that cyclophilin B (CypB) expression is important for malignant glioblastoma multiforme (GBM) cell proliferation. To identify molecular mechanisms that could explain CypB-dependent survival in human GBM cells, a microarray analysis was performed using RNA prepared from U251MG GBM cells transduced with lentiviral CypB shRNA. These data revealed that about 130 genes were more than 2-fold affected by CypB depletion. Significant alterations in the expression of genes related to cell death, cell proliferation and cell migration were found in the shCypB cells. U251 glioblastoma cells transduced with lentivirus expressing non-target control shRNA (shCON) were compared to cells transduced with lentivirus expressing shRNA sequence against cyclophilin B (shCypB). Cells transduced with the lentiviral shRNA (shCON VS shCypB) for 5 days before total RNA extraction. Three biological replicates of cells treated with each shRNA (shCON or shCypB) were profiled.