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Expression of miRNAs in pancreatic adenocarcinoma-relationship to morphological characteristics and disease prognosis.
Szabó, Arpád ; Mandys, Václav (advisor) ; Hermanová, Markéta (referee) ; Daum, Ondřej (referee)
Ductal adenocarcinoma (PDAC) is the most common primary pancreatic neoplasm. It's frequently diagnosed in late inoperable stage and has high resistance to chemotherapy; this situation contributes to its unfavourable prognosis. Therefore, there is a need for biomarkers enabling early detection of PDACs and by this way to improve patient prognosis. MicroRNAs (miRNA), short non-coding RNA molecules involved in post-transcriptional regulation of gene expression, belong to such markers. Contrary to other RNA molecules, miRNAs are stable in biological samples. Their expression is measured by several analytical methods, including real-time quantitative PCR (RT-qPCR). Normalization of methods determining miRNA levels requires adequate endogenous controls. However, variable expression of endogenous controls in tumors may cause bias in determining miRNA levels. The aim of the first study was to investigate the expression of six miRNAs isolated from formalin fixed paraffin embedded (FFPE) samples of PDACs. Four controls were chosen for RT-qPCR result normalization: artificial spike miR-39 from C. elegans, U6 snRNA, miR-16 and snoRNA U91. Expression values of all studied miRNAs in tumors were significantly different depending on selected endogenous controls. Additionally, stability of the controls varied...
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