5-carboxytetramethylrhodamine-succinimidyl-ester and Carcinoma--Krebs-2

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.

Topics: Alu Elements; Animals; Biomarkers, Tumor; Carcinoma, Krebs 2; Cell Differentiation; DNA; Fluorescent Dyes; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Gene Regulatory Networks; High-Throughput Nucleotide Sequencing; Neoplastic Stem Cells; Phenotype; Real-Time Polymerase Chain Reaction; Rhodamines; Sequence Analysis, RNA; Signal Transduction; Transcriptome