miRNA as markers for the diagnostic screening of colon cancer.

Early screening for colon cancer (CC) allows for early stage diagnosis of the malignancy and potentially reduces disease mortality as the cancer is most likely curable at its earliest stages. Early detection would be desirable if accurate, practical and cost-effective diagnostic measures for this cancer were available. Mortality and morbidity from CC represent a major health problem involving a malignant disease that is theoretically preventable through screening. Current screening methods (e.g., the convenient and inexpensive immunological fecal occult blood test, FOBTi, obtained from patients' medical records) either lack sensitivity and require dietary restriction, which impedes compliance and use; are costly (e.g., colonoscopy), which decreases compliance; or could result in mortality. In comparison with the FOBT test, a non-invasive sensitive screen for which there is no requirement for dietary restriction would be a more convenient test. Colorectal cancer is the only cancer for which colonoscopy is recommended as a screening method. Although colonoscopy is a reliable screening tool, the invasive nature, abdominal pain, potential complications and high cost have hampered the application of this procedure worldwide. A screening approach using the stable miRNA molecules, which are relatively non-degradable when extracted from non-invasive stool and semi-invasive blood samples by commercially available kits and manipulated thereafter, would be preferable to a transcriptomic mRNA-, a mutation DNA-, an epigenetic- or a proteomic-based test. The approach uses reverse transcriptase, modified real-time quantitative PCR. Although exosomal RNA would be missed, using a restricted extraction of total RNA from stool or blood, a parallel test could also be carried out on RNA obtained from stool or plasma samples, and appropriate corrections for exsosomal loss can be made for accurate and quantitative test result. Eventually, a chip can be developed to facilitate diagnosis, as has been done for the quantification of genetically modified organisms in foods. The gold standard to which the molecular miRNA test is compared is colonoscopy, which can be obtained from patients' medical records. If performance criteria are met, as detailed herein, a miRNA test in human stool or blood samples based on high-throughput automated technologies and quantitative expression measurements commonly used in the diagnostic clinical laboratory should be advanced to the clinical setting, which will make a significant impact on CC prevention.