Identification and Validation of Reference Genes for qPCR Detection of Serum microRNAs in Colorectal Adenocarcinoma Patients

• DOI: 10.1371/journal.pone.0083025
• Publication Date: 2013-12-11T16:34:42Z
• Authors: Guixi Zheng; Haiyan Wang; Xin Zhang; Yongmei Yang; Lili Wang; Lutao Du; Wei Li; Juan Li; Ailin Qu; Yimin Liu; Chuanxin Wang
• Keywords: Reference Genes

Serum microRNAs (miRNAs) have become a highlighted research hotspot, especially for their great potential as novel promising non-invasive biomarker in cancer diagnosis. The most frequently used approach serum miRNAs detection is quantitative real time polymerase chain reaction (qPCR). In order to obtain reliable qPCR data of expression, the use reference genes endogenous control undoubtly necessary. However, no systematic evaluation and validation normalizing analysis has been reported colorectal adenocarcinoma. We firstly profiled pooled adenocarcinoma, adenoma healthy controls selected list 13 candidate genes. U6 snRNA (U6) above-mentioned were included further confirmation by qPCR. As result, 5 (miR-151a-3p, miR-4446-3p, miR-221-3p, miR-93-5p miR-3184-3p) not detected all samples 2 (miR-197-3p miR-26a-5p) relatively low with median Cq more than 35, excluded from stability analysis. Then variable other 6 (miR-103b, miR-484, miR-16-5p, miR-3615, miR-18a-3p miR-191-5p) evaluated using two algorithms: geNorm NormFinder which both identified miR-191-5p stably expressed gene stable pair After validating an independent large cohorts selecting miR-92a-3p target miRNA evaluate effect gene, we propose that combination could be controls.