BioProject

In our study, we proposed first to reduce the profiling variability by selecting the miRNA isolation kit offering the higher input RNA concentration for microarray profiling and second to control the profiling step variability by adapting the widely used exogenous normalization of the validation step to the profiling one. More...

In our study, we proposed first to reduce the profiling variability by selecting the miRNA isolation kit offering the higher input RNA concentration for microarray profiling and second to control the profiling step variability by adapting the widely used exogenous normalization of the validation step to the profiling one. The new strategy proposed could improve the overlap between independent studies using microarray profiling, but also potentially with other high-throughput methods, to finally identify pertinent miRNA signatures in biofluids. Because this project is only a methodological study, the cohort of serum and plasma samples used in this study were all obtained from patients with FIGO stages III/IV serous ovarian cancer. This study was approved by the ethics committee Nord-Ouest III [CPP N° 2011-02] and national authorities (AFSSAPS N°B110260-20). Plasma samples were obtained from the Comprehensive Cancer Center F. Baclesse biological collections that have been declared (# DC-2010-1243) to the French Ministry of Research and Superior Education. Prior to scientific use of samples and data, patients were appropriately informed and asked to consent in writing, in compliance with French and European regulations. Overall design: 13 samples of serum from patients with homogeneous clinical status were used to purify circulating miRNAs. Purified miR levels were assessed by qPCR and by hybridization on Affymetrix miRNA V4 arrays. Arrays data were normalized by different methods. Comparison between the data from qPCR and arrays were performed to identify the best normalization approach. Less...