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Oncotarget ; 8(6): 9425-9441, 2017 Feb 07.
Article in English | MEDLINE | ID: mdl-28031533

ABSTRACT

Using the ability of poorly differentiated cells to natively internalize fragments of extracellular double-stranded DNA as a marker, we isolated a tumorigenic subpopulation present in Krebs-2 ascites that demonstrated the features of tumor-inducing cancer stem cells. Having combined TAMRA-labeled DNA probe and the power of RNA-seq technology, we identified a set of 168 genes specifically expressed in TAMRA-positive cells (tumor-initiating stem cells), these genes remaining silent in TAMRA-negative cancer cells. TAMRA+ cells displayed gene expression signatures characteristic of both stem cells and cancer cells. The observed expression differences between TAMRA+ and TAMRA- cells were validated by Real Time PCR. The results obtained corroborated the biological data that TAMRA+ murine Krebs-2 tumor cells are tumor-initiating stem cells. The approach developed can be applied to profile any poorly differentiated cell types that are capable of immanent internalization of double-stranded DNA.


Subject(s)
Biomarkers, Tumor/genetics , Carcinoma, Krebs 2/genetics , Cell Differentiation , Gene Expression Profiling/methods , Transcriptome , Alu Elements , Animals , Biomarkers, Tumor/metabolism , Carcinoma, Krebs 2/pathology , DNA/genetics , DNA/metabolism , Fluorescent Dyes/metabolism , Gene Expression Regulation, Neoplastic , Gene Regulatory Networks , High-Throughput Nucleotide Sequencing , Neoplastic Stem Cells/metabolism , Neoplastic Stem Cells/pathology , Phenotype , Real-Time Polymerase Chain Reaction , Rhodamines/metabolism , Sequence Analysis, RNA , Signal Transduction
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