Measurement of miRNAs in Chronic Lymphocytic Leukemia Patient Samples by Quantitative Reverse Transcription PCR

MicroRNAs (miRNAs) are small noncoding RNAs that target specific mRNAs through interaction with complementary sequences usually found in the 3'-untranslated regions (UTRs) of target mRNAs. miRNAs have been shown to play a fundamental role in the management of chronic lymphocytic leukemia (CLL) by modulating gene expression patterns and cellular signaling pathways. In recent years, several studies have focused on the role of regulatory miRNAs in the pathogenesis of CLL. Aberrant expression of CLL-specific miRNAs has emerged as therapeutic and diagnostic biomarkers in patients with CLL. Here, we describe a method for the quantification of miRNAs in malignant B cells from the mononuclear cell compartment, isolated from peripheral blood. We focus on the isolation of human blood monocytes by Ficoll-Paque gradient centrifugation, total RNA extraction from human peripheral blood mononuclear cells, and quantitative reverse transcription (qRT)-PCR, which is useful for the measurement of miRNAs in monocytes isolated from blood samples.