MicroRNA profiling analysis of differences between bone marrow mouse stem cells versus leukemic and teratocarcinomic cell lines

Our aim is to identify any hallmark differentially expressed miRNAs between mouse bone marrow cells and different cancer cell lines (leukemic with lymphoid and myeloid origin and teratocarcinomic with dedifferentiated characteristics). MicroRNAs (miRNAs) are an important member of posttrascriptional regulators and its pathological level changes have been implicated in various cancer types. This is the first step of the study focused at cancer stem cell biology of cancer in our in vivo mouse models. We conclude by high-throughput microRNA profiling analysis,that exists a group of miRNAs candidates to explore their biological function in mice bone marrow stem cells an its role in carcinogenesis. We isolated the bone marrow cells from mice by femur and tibial flush, then wash and collected. We isolate the same number of cells from fresh cultures of different cell lines types at passage 3, then wash and collected. Total RNA was extracted for the TaqMan Low Density Array v2.0.RNA quality was assessed using Nanodrop for concentration and purity (OD 260/280 and 260/230) and the Agilent Bioanalyzer 2100 for integrity (RNA integrity number (RIN) > 9). Normalization was performed with common endogenous controls for A card and B card (MummU6, snoRNA135, snoRNA202 reference snRNAs for mouse).Comparative real-time PCR was performed in 3 replicates, including no-template controls.