Summary: | <b>Introduction:</b> One-third of colorectal cancer (CRC) patients present with advanced disease, and establishing control remains a challenge. Identifying novel biomarkers to facilitate earlier diagnosis is imperative in enhancing oncological outcomes. We aimed to create miRNA oncogenic signature to aid CRC diagnosis. <b>Methods</b>: Tumour and tumour-associated normal (TAN) were extracted from 74 patients during surgery for CRC. RNA was isolated and target miRNAs were quantified using real-time reverse transcriptase polymerase chain reaction. Regression analyses were performed in order to identify miRNA targets capable of differentiating CRC from TAN and compared with two endogenous controls (miR-16 and miR-345) in each sample. Areas under the curve (AUCs) in Receiver Operating Characteristic (ROC) analyses were determined. <b>Results</b>: MiR-21 (β-coefficient:3.661, SE:1.720, <i>p</i> = 0.033), miR-31 (β-coefficient:2.783, SE:0.918, <i>p</i> = 0.002), and miR-150 (β-coefficient:−4.404, SE:0.526, <i>p</i> = 0.004) expression profiles differentiated CRC from TAN. In multivariable analyses, increased miR-31 (β-coefficient:2.431, SE:0.715, <i>p</i> < 0.001) and reduced miR-150 (β-coefficient:−4.620, SE:1.319, <i>p</i> < 0.001) independently differentiated CRC from TAN. The highest AUC generated for miR-21, miR-31, and miR-150 in an oncogenic expression assay was 83.0% (95%CI: 61.7–100.0, <i>p</i> < 0.001). In the circulation of 34 independent CRC patients and 5 controls, the mean expression of miR-21 (<i>p</i> = 0.001), miR-31 (<i>p</i> = 0.001), and miR-150 (<i>p</i> < 0.001) differentiated CRC from controls; however, the median expression of miR-21 (<i>p</i> = 0.476), miR-31 (<i>p</i> = 0.933), and miR-150 (<i>p</i> = 0.148) failed to differentiate these groups. <b>Conclusion</b>: This study identified a five-miRNA signature capable of distinguishing CRC from normal tissues with a high diagnostic test accuracy. Further experimentation with this signature is required to elucidate its diagnostic relevance in the circulation of CRC patients.
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