Abstract

Salivary microRNAs (miRNAs) are of high interest as diagnostic biomarkers for non-oral cancer. However, little is known about their value for colorectal cancer (CRC) detection. Our study aims to characterize salivary miRNAs in order to identify non-invasive markers for CRC diagnosis. The screening of 754 miRNAs was performed in saliva samples from 14 CRC and 10 healthy controls. The differential expressed miRNAs were validated by RT-qPCR in 51 CRC, 19 adenomas and 37 healthy controls. Receiver operating characteristic (ROC) curves and logistic regression models were performed to analyze the clinical value of these miRNAs. Twenty-two salivary miRNAs were significantly deregulated in CRC patients vs. healthy individuals (p < 0.05) in the discovery phase. From those, five upregulated miRNAs (miR-186-5p, miR-29a-3p, miR-29c-3p, miR-766-3p, and miR-491-5p) were confirmed to be significantly higher in the CRC vs. healthy group (p < 0.05). This five-miRNA signature showed diagnostic value (72% sensitivity, 66.67% specificity, AUC = 0.754) to detect CRC, which was even higher in combination with carcinoembryonic antigen (CEA) levels. Overall, after the first global characterization of salivary miRNAs in CRC, a five-miRNA panel was identified as a promising tool to diagnose this malignancy, representing a novel approach to detect cancer-associated epigenetic alterations using a non-invasive strategy.

Highlights

  • Colorectal cancer (CRC) is the third most commonly diagnosed cancer in men and the second in women worldwide

  • This study was designed as two phases: a discovery phase including 24 saliva samples (10 healthy controls and 14 CRC, including 7 non-metastatic and 7 metastatic patients) and a validation phase comprising 107 saliva samples (37 healthy controls, 19 adenomas and 51 CRC), including those used for the discovery phase (Figure S1)

  • 754 miRNAs were profiled in saliva samples from 14 CRC patients and 10 healthy controls

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Summary

A Novel Saliva-Based miRNA Signature for Colorectal Cancer Diagnosis

Óscar Rapado-González 1,2,3 , Blanca Majem 4,5, Ana Álvarez-Castro 6 , Roberto Díaz-Peña 2,7 , Alicia Abalo 2, Leticia Suárez-Cabrera 4, Antonio Gil-Moreno 3,8 , Anna Santamaría 4 , Rafael López-López 3,9 , Laura Muinelo-Romay 2,3,* and María Mercedes Suarez-Cunqueiro 1,10,*.

Introduction
Study Subjects and Sample Collection
Saliva RNA Extraction
Salivary MiRNA Discovery Profiling with TaqMan Low-Density Arrays
MiRNA Expression Analysis Using RT-qPCR
Functional Enrichment Analysis
Statistical Analysis
Results
A Panel of Salivary miRNAs Was Validated for CRC
Functional Annotation Analysis of Salivary miRNAs
Conclusions
Full Text
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