MicroRNA expression profile analysis in human skeletal muscle tissue: Selection of critical reference - 29/04/23
Abstract |
MicroRNAs (miRNAs) are highly conserved small non-coding RNAs, that modulate gene expression by targeting messenger RNA of many processes. Thus, miRNAs are key regulators of both physiological and pathological settings. Reliable results of quantitative miRNA evaluation depend on suitable reference genes (RGs) for data normalization. To date, no consensus has been reached on the best RG for muscle tissue. We assessed RGs stability in skeletal muscle tissue in patients with spinal deformity. Ninety tissue samples were obtained from the deep paravertebral muscles from the convex and concave sides of the spinal curvature, as well as the superficial paraspinal muscles. We evaluated the stability of twelve miRNAs (hsa-miR-1–3p, hsa-miR-1–5p, hsa-miR-26b-5p, hsa-miR-92a-3p, hsa-miR-133a-3p, hsa-miR-133a-5p, hsa-miR-133b, hsa-miR-191–5p, hsa-miR-206, hsa-miR-208b-5p, hsa-miR-486–5p, hsa-miR-499a-5p), finding three to be indicative of reference miRNA, and nine as muscle-tissue specific. Stability was quantified using four statistical tools and a comprehensive ranking system. Three miRNAs were indicated as the most stable, and we assessed hsa-miR-486–5p as the most, and hsa-miR-208b-5p as the least suitable RGs for miRNA quantitative analyses. We recommend using a minimum of three RGs miRNA to normalize RT-qPCR data. Finally, qPCR efficiency should always be considered. To obtain consistent results, data normalization in muscle tissue is required.
Le texte complet de cet article est disponible en PDF.Graphical Abstract |
Highlights |
• | Quantitative real-time PCR (qPCR) data normalization in muscle tissue is required. |
• | Quantitative miRNA expression results depend on reference genes for data normalization. |
• | qPCR efficiency should always be considered. |
• | We recommend using at least three reference miRNA genes to normalize quantitative data. |
Keywords : Reference microRNA (ref miRNA), Muscle tissue, MicroRNA stability, Quantitative polymerase chain reaction normalization
Plan
Vol 162
Article 114682- juin 2023 Retour au numéroBienvenue sur EM-consulte, la référence des professionnels de santé.
L’accès au texte intégral de cet article nécessite un abonnement.
Déjà abonné à cette revue ?