MiRNA Analysis by Quantitative PCR in Preterm Human Breast Milk Reveals Daily Fluctuations of hsa-miR-16-5p

• DOI: 10.1371/journal.pone.0140488
• Publication Date: 2015-10-16T18:18:38Z
• Authors: Ilaria Floris; Hélène Billard; Clair-Yves Boquien; Evelyne Joram-Gau...; Laure Simon; Arnaud Legrand; Cécile Boscher; Jean-Christophe Rozé; Francisco Bolaños...; Bertrand Kaeffer
• Keywords: expression génique; composition chimique; [SDV]Life Sciences [q-bio]; Science; pcr quantitative; 610; Real-Time Polymerase Chain Reaction; micro rna; fluctuation diurne; 03 medical and health sciences; Humans; 2. Zero hunger; 0303 health sciences; Milk, Human; Q; R; Circadian Rhythm; [SDV] Life Sciences [q-bio]; MicroRNAs; lait de femme; Medicine; variation qualitative; Female; Infant, Premature; Research Article

Background and Aims Human breast milk is an extremely dynamic fluid containing many biologically-active components which change throughout the feeding period day. We designed a miRNA assay on minimized amounts of raw obtained from mothers preterm infants. investigated changes in expression within month 2 lactation then over course 24 hours. Materials Methods Analyses were performed pooled milk, made by combining samples collected at different clock times same mother donor, along with time series hours four unsynchronized mothers. Whole lipids or skim fractions processed analyzed qPCR. measured hsa-miR-16-5p, hsa-miR-21-5p, hsa-miR-146-5p, hsa-let-7a, d g (all -5p). Stability endogenous controls was evaluated using RefFinder, web tool integrating geNorm, Normfinder, BestKeeper comparative ΔΔCt method. Results MiR-21 miR-16 stably expressed whole Analysis revealed that miR-146b let-7d better references both fractions. Time (5H-23H) allowed identification set three reference genes (hsa-let-7d, hsa-let-7g miR-146b) to normalize quantification cycle (Cq) data. identified daily oscillation miR-16-5p. Perspectives Our allows exploring levels baby 48–72