RESUMEN

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.

Asunto(s)

Biomarcadores de Tumor/genética , Carcinoma Krebs 2/genética , Diferenciación Celular , Perfilación de la Expresión Génica/métodos , Transcriptoma , Elementos Alu , Animales , Biomarcadores de Tumor/metabolismo , Carcinoma Krebs 2/patología , ADN/genética , ADN/metabolismo , Colorantes Fluorescentes/metabolismo , Regulación Neoplásica de la Expresión Génica , Redes Reguladoras de Genes , Secuenciación de Nucleótidos de Alto Rendimiento , Células Madre Neoplásicas/metabolismo , Células Madre Neoplásicas/patología , Fenotipo , Reacción en Cadena en Tiempo Real de la Polimerasa , Rodaminas/metabolismo , Análisis de Secuencia de ARN , Transducción de Señal