Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Transcription profiling of human mesenchymal stem cells transformed with lentiviral vectors of TERT, E6, SV40 and RAS

ABSTRACT: Mesenchymal Stem Cells are transformed by step-wise stable transfection with lentiviral vectors of TERT, E6, E7, SV40, and Ras.

ORGANISM(S): Homo sapiens

SUBMITTER: Stephen Henderson

PROVIDER: E-MEXP-563 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Transformation of human mesenchymal stem cells increases their dependency on oxidative phosphorylation for energy production.

Funes Juan M JM Quintero Marisol M Henderson Stephen S Martinez Dolores D Qureshi Uzma U Westwood Claire C Clements Mark O MO Bourboulia Dimitra D Pedley R Barbara RB Moncada Salvador S Boshoff Chris C

Proceedings of the National Academy of Sciences of the United States of America 20070323 15

An increased dependency on glycolysis for ATP production is considered to be a hallmark of tumor cells. Whether this increase in glycolytic activity is due mainly to inherent metabolic alterations or to the hypoxic microenvironment remains controversial. Here we have transformed human adult mesenchymal stem cells (MSC) using genetic alterations as described for differentiated cells. Our data suggest that MSC require disruption of the same pathways as have been shown for differentiated cells to c ...[more]

PMID: 17384149

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Project description:Most adult patients have a D816V mutation in phosphotransferase domain (PTD), we have described that half of the children carry mutations in extracellular domain (ECD). KIT-ECD versus IT-PTD-mutants were introduced into rodent Ba/F3, EML, Rat2 and human TF1 cells to investigate their biological effect. ECD- and PTD-mutants also displayed distinct whole-genome transcriptional profiles in EML cells. We observed differences in their signaling properties: they both activated STAT pathways, whereas AKT pathway was only activated by ECD-mutants. Consistently, AKT inhibitor suppressed ECD-mutant-dependent proliferation, clonogenicity and erythroid differentiation. Expression of myristoylated AKT restored erythroid differentiation in EMLPTD cells, suggesting the differential role of AKT in those mutants. Overall, our study implied different pathogenesis of pediatric versus adult mastocytosis, which might explain their diverse phenotypes. Each EML cell line (Del417-419insY mutant and D816V mutant) was cultured with or without 250 ng/mL SCF for 48h. Gene expression profile analysis was performed using whole-genome microarrays. RNA expression profiling of cell lines was done with Affymetrix M430 2.0 mouse oligonucleotide microarrays containing 45.101 probe sets, representing 21.408 transcripts and variants including 17.482 well-characterized mouse genes. Preparation of cRNA, hybridizations, washes, detection and quantification were done as recommended by the supplier. For each sample, synthesis of the first-strand cDNA was done from 3 μg total RNA by T7-oligo(dT) priming, followed by second-strand cDNA synthesis. After purification, in vitro transcription associated with amplification generated cRNA-containing biotinylated pseudouridine. Biotinylated cRNA was purified, quantified and chemically fragmented (95°C for 35 min), then hybridized to microarrays in 200 μL hybridization buffer at 45°C for 16 h. Automated washes and staining with streptavidin-phycoerythrin were done as recommended. Signal amplification was done by biotinylated antistreptavidin antibody with goat-IgG blocking antibody. Scanning was done with Affymetrix GeneArray scanner and quantification with Affymetrix GCOS software.

Dataset Information

Transcription profiling of human mesenchymal stem cells transformed with lentiviral vectors of TERT, E6, SV40 and RAS

Publications

Transformation of human mesenchymal stem cells increases their dependency on oxidative phosphorylation for energy production.

Similar Datasets

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