Novel microRNAs modulating ecto-5′-nucleotidase expression [MDA-MB-231]. Novel microRNAs modulating ecto-5′-nucleotidase expression [MDA-MB-231]

the effect of NT5E enhancing miRNAs was studied by gene expression profiling to unterstand mechanism of observed NT5E upregulation. Overall design: Gene expression profiling of tumor cell lines transfected with miRNAs enhancing NT5E expression was performed as follows: MDA-MB-231 cells were transfected with 50 nM miRNA, including a non-targeting control miRNA (control-1/ath-miR-416) in a 12 well plate format, performing triplicates for each format. After 48 h, cells were harvested for RNA isolation using an RNAeasy kit (Qiagen, Hilden, Germany). Microarrays were performed by DKFZ-Genomics and Proteomics Core Facility using an Affymetrix Clariom S human chip for all samples. Raw data processing was also performed using the Core Facility. Differential gene expression was calculated by comparing miRNA-induced gene expression levels to the respective gene expression levels following mimic control-1 transfection.