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J Clin Pathol ; 66(3): 249-52, 2013 Mar.
Article in English | MEDLINE | ID: mdl-23172553

ABSTRACT

Tissue biopsy specimens in the form of formalin-fixed paraffin-embedded tissue (FFPET) represent a valuable resource for biomarker identification and validation. However, to date, they remain an underused asset due to uncertainty regarding RNA extraction and the reliability of downstream techniques, including quantitative RT-PCR. Recently, much interest has emerged in the study of microRNAs; small single-stranded RNAs with a role in transcriptional regulation, that are thought to be well preserved in FFPET. In this study, we show that microRNA expression is comparable between FFPET and matched fresh-frozen samples (miR-17-5p: p=0.01, miR-92: p=0.003), and demonstrate that no significant deterioration in expression occurs over prolonged FFPET storage (p=0.06). Furthermore, microRNA expression is equivalent dependant on RNA extraction method (p<0.001) or DNAse treatment of total RNA (p<0.001). Finally, we validate miR-24 as a suitable reference microRNA for diffuse large B-cell lymphoma (DLBCL) FFPET studies.


Subject(s)
Gene Expression Profiling , Genetic Markers/genetics , Lymphoma, Large B-Cell, Diffuse/genetics , MicroRNAs/genetics , Paraffin Embedding , Tissue Fixation/methods , Biomarkers, Tumor/genetics , Biopsy , Cohort Studies , Fixatives , Formaldehyde , Humans , Lymphoma, Large B-Cell, Diffuse/pathology , Reagent Kits, Diagnostic , Reference Standards , Reproducibility of Results , Reverse Transcriptase Polymerase Chain Reaction
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