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We acknowledge the funding from Advanced Marker Discovery (AMADIX) and grants support from Fundacion Cientifica de la Asociacion Espanola contra el Cancer (GCB13131592CAST), Ministerio de Economia y Competitividad (RTC-2015-3850-1 and SAF2014-54453-R) and Instituto de Salud Carlos III (PI17/01003, co-funded by FEDER-European Union). CIBEREHD is funded by the Instituto de Salud Carlos III. We are grateful to the support of CERCA Programme and SGR 653 from Generalitat de Catalunya. This work was developed at the Centro Esther Koplowitz, Barcelona, Spain.

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Validation of miR-1228-3p as Housekeeping for MicroRNA Analysis in Liquid Biopsies from Colorectal Cancer Patients

Publicat a:Biomolecules. 10 (1): E16- - 2020-01-01 10(1), DOI: 10.3390/biom10010016

Autors: Duran-Sanchon, Saray; Vila-Navarro, Elena; Marcuello, Maria; Jose Lozano, Juan; Munoz, Jenifer; Cubiella, Joaquin; Soledad Diez, Maria; Bujanda, Luis; Lanas, Angel; Jover, Rodrigo; Hernandez, Vicent; Quintero, Enrique; Herreros-Villanueva, Marta; Carmen Martin, Ana; Perez-Palacios, Rosa; Arroyo, Rocio; Castells, Antoni; Gironella, Meritxell

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Circulating microRNA (miRNA) analysis is a growing research field. However, it usually requires an endogenous control or housekeeping (HK) in order to normalize expression of specific miRNAs throughout different samples. Unfortunately, no adequate HK for circulating miRNA analysis is still known in the colorectal cancer (CRC) context whereas several have been suggested. Hence, our aims were to validate the previously suggested miR-1228-3p as HK for CRC studies, to compare its suitability with the widely used miR-16-5p, and to evaluate the influence of hemolysis on both miRNAs.We analyzed by quantitative reverse-transcription polymerase chain reaction (qRT-PCR) the expression of miR-1228-3p, miR-16-5p and the spike-in cel-miR-39 in a set of 297 plasmas (92 CRC, 101 advanced adenomas -AA-, and 100 controls) and 213 serum samples (59 CRC, 74 AA and 80 controls). We also analyzed both miRNAs depending on the hemolysis degree in 7 plasmas and 31 serums.Levels of miR-1228-3p and miR-16-5p did not show significant differences between groups although miR-16-5p exhibited more variability in plasma and serum samples. Importantly, the combination of cel-miR-39 and miR-1228-3p was the most stable one. Moreover, we observed that miR-16-5p was significantly influenced by hemolysis in contrast with miR-1228-3p that exhibited no correlation with this confounding factor in both biofluids.MiR-1228-3p has been validated as an adequate endogenous control for circulating miRNA analysis in CRC and AA liquid biopsies.

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