Establishment of Robust Controls for the Normalization of MiRNA Expression in Neuroendocrine Tumors of the Ileum and Pancreas
Overview
Affiliations
There is need to determine tissue-specific robust controls for normalization of microRNA expression to avoid false results and misinterpretation. The aim of this study was to evaluate the expression of different small RNAs in neuroendocrine tumors (NETs) and their suitability as normalizers in miRNA real-time PCR experiments. We investigated the expression of the nine small RNAs miR-93, miR-191, SNORD48, SNORD61, SNORD68, SNORD72, SNORD95, SNORD96a, and RNU6-2 in formalin-fixed, paraffin-embedded tissue samples of 25 ileal NETs by real-time PCR determining the most stable controls for expression normalization using four different algorithms. This analysis was expended to ten pancreatic NETs. Finally, five small RNAs were further tested as normalizers for miRNA-133a expression, which is known to be downregulated in metastases of ileal NETs, in ten matched pairs of ileal NETs and their metastases. Ranking of the expression results revealed the following order of stability from high to low: SNORD61 < SNORD95 < SNORD72 < SNORD96a < SNORD68 < miR-191 < miR-93 < RNU6-2 < SNORD48 for ileal NETs and SNORD95 < miR-93 < SNORD96a < SNORD61 < SNORD68 < SNORD72 < RNU6-2 < miR-191 < SNORD48 for pancreatic NETs. The determination of SNORD61 and SNORD95 for ileal NETs and SNORD95 and miR-93 for pancreatic NETs as good normalizers presents a useful tool for experiments involving the analysis of miRNA expression.
Blazquez-Encinas R, Moreno-Montilla M, Garcia-Vioque V, Gracia-Navarro F, Alors-Perez E, Pedraza-Arevalo S Rev Endocr Metab Disord. 2022; 24(2):267-282.
PMID: 36418657 PMC: 9685014. DOI: 10.1007/s11154-022-09771-4.
Latini A, Vancheri C, Amati F, Morini E, Grelli S, Matteucci C J Cell Mol Med. 2022; 26(19):4940-4948.
PMID: 36073344 PMC: 9538662. DOI: 10.1111/jcmm.17492.
Buglyo G, Magyar Z, Romicsne Gorbe E, Banusz R, Csoka M, Micsik T Med Sci Monit. 2021; 27:e932731.
PMID: 34608109 PMC: 8501895. DOI: 10.12659/MSM.932731.
Extracellular Vesicle-Based Detection of Pancreatic Cancer.
Verel-Yilmaz Y, Fernandez J, Schafer A, Nevermann S, Cook L, Gercke N Front Cell Dev Biol. 2021; 9:697939.
PMID: 34368146 PMC: 8343017. DOI: 10.3389/fcell.2021.697939.
An explorative study identifies miRNA signatures for the diagnosis of non-celiac wheat sensitivity.
Clemente E, Efthymakis K, Carletti E, Capone V, Sperduti S, Bologna G PLoS One. 2019; 14(12):e0226478.
PMID: 31834915 PMC: 6910677. DOI: 10.1371/journal.pone.0226478.