OCR Text |
Show 219 THE UNIVERSITY OF UTAH Primary sclerosing cholangitis (PSC) and ulcerative colitis (UC) are diseases that target the GI tract. The causes of these diseases are generally unknown. However, if these diseases remain untreated and are allowed to continually progress, they may develop into cholongiocarcino-ma (CCA), a cancer found in the bile ducts. The primary objective of this study was to observe microRNAs (miRNAs) and their gene expression levels in PSC/UC versus CCA. Differences in the miRNA expression between PSC/UC and CCA in patient serum can alert to changes in the current status of the disease. Additionally these miRNAs can serve as GI disease biomarkers, playing an important role in early cancer detection and evaluating potential risks for develop-ing CCA. Serum and tissue samples were obtained from two PSC/UC patients and two CCA patients. The miRNA was extracted from these samples and submitted to the Agilent miRNA microarray to distinguish a specific subset of miRNAs exhibiting differential expression between the two groups. From this information, 34 miRNA primers were developed using the exact miRNA se-quence of interest. Several factors, including sequence length, GC content, melting tempera-ture, and primer stability were analyzed before reaching the optimal primer sequence. Due to the characteristic short sequences of miRNAs, Rapid Amplification of cDNA Ends (RACE) PCR was used to extend the length and to convert the Stratagene Universal Human Reference RNA to cDNA. The primers were then submitted to the Roche LightCycler 480 for quantitative Real Time (qRT) PCR to verify primer efficiency. The results are currently pending, as the primers are in the process of being validated. After primer development and verification, more miRNA samples from patient sera will be collected and analyzed using qRT-PCR. The implications of this study suggest that a blood test could be used as an early detection tool that is not only less invasive than that of a biopsy and but also less daunting to the patient. PRIMER DESIGN FOR Q-PCR OF MICRORNA FROM SERUM FOR THE DEVELOPMENT OF GI DISEASE BIOMARKERS Irene Pan (Phil Bernard) Department of Pathology University of Utah health sciences leap program Irene Pan Phil Bernard |