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

Katrien Van Roosbroeck, Recep Bayraktar, George A Calin

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

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.

LanguageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages267-276
Number of pages10
DOIs
StatePublished - Jan 1 2019

Publication series

NameMethods in Molecular Biology
Volume1881
ISSN (Print)1064-3745

Fingerprint

B-Cell Chronic Lymphocytic Leukemia
MicroRNAs
Reverse Transcription
Polymerase Chain Reaction
Monocytes
Small Untranslated RNA
Ficoll
Messenger RNA
3' Untranslated Regions
Centrifugation
Blood Cells
B-Lymphocytes
Biomarkers
RNA
Gene Expression

Keywords

  • Chronic lymphocytic leukemia
  • Ficoll-Paque density gradient
  • microRNA
  • Peripheral blood mononuclear cells
  • Quantitative reverse transcription (qRT)-PCR

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Van Roosbroeck, K., Bayraktar, R., & Calin, G. A. (2019). Measurement of miRNAs in Chronic Lymphocytic Leukemia Patient Samples by Quantitative Reverse Transcription PCR. In Methods in Molecular Biology (pp. 267-276). (Methods in Molecular Biology; Vol. 1881). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-8876-1_20

Measurement of miRNAs in Chronic Lymphocytic Leukemia Patient Samples by Quantitative Reverse Transcription PCR. / Van Roosbroeck, Katrien; Bayraktar, Recep; Calin, George A.

Methods in Molecular Biology. Humana Press Inc., 2019. p. 267-276 (Methods in Molecular Biology; Vol. 1881).

Research output: Chapter in Book/Report/Conference proceedingChapter

Van Roosbroeck, K, Bayraktar, R & Calin, GA 2019, Measurement of miRNAs in Chronic Lymphocytic Leukemia Patient Samples by Quantitative Reverse Transcription PCR. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 1881, Humana Press Inc., pp. 267-276. https://doi.org/10.1007/978-1-4939-8876-1_20
Van Roosbroeck K, Bayraktar R, Calin GA. Measurement of miRNAs in Chronic Lymphocytic Leukemia Patient Samples by Quantitative Reverse Transcription PCR. In Methods in Molecular Biology. Humana Press Inc. 2019. p. 267-276. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-4939-8876-1_20
Van Roosbroeck, Katrien ; Bayraktar, Recep ; Calin, George A. / Measurement of miRNAs in Chronic Lymphocytic Leukemia Patient Samples by Quantitative Reverse Transcription PCR. Methods in Molecular Biology. Humana Press Inc., 2019. pp. 267-276 (Methods in Molecular Biology).
@inbook{0cfa3034708542c78854cf4c114216e8,
title = "Measurement of miRNAs in Chronic Lymphocytic Leukemia Patient Samples by Quantitative Reverse Transcription PCR",
abstract = "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.",
keywords = "Chronic lymphocytic leukemia, Ficoll-Paque density gradient, microRNA, Peripheral blood mononuclear cells, Quantitative reverse transcription (qRT)-PCR",
author = "{Van Roosbroeck}, Katrien and Recep Bayraktar and Calin, {George A}",
year = "2019",
month = "1",
day = "1",
doi = "10.1007/978-1-4939-8876-1_20",
language = "English (US)",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "267--276",
booktitle = "Methods in Molecular Biology",

}

TY - CHAP

T1 - Measurement of miRNAs in Chronic Lymphocytic Leukemia Patient Samples by Quantitative Reverse Transcription PCR

AU - Van Roosbroeck, Katrien

AU - Bayraktar, Recep

AU - Calin, George A

PY - 2019/1/1

Y1 - 2019/1/1

N2 - 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.

AB - 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.

KW - Chronic lymphocytic leukemia

KW - Ficoll-Paque density gradient

KW - microRNA

KW - Peripheral blood mononuclear cells

KW - Quantitative reverse transcription (qRT)-PCR

UR - http://www.scopus.com/inward/record.url?scp=85055174412&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85055174412&partnerID=8YFLogxK

U2 - 10.1007/978-1-4939-8876-1_20

DO - 10.1007/978-1-4939-8876-1_20

M3 - Chapter

T3 - Methods in Molecular Biology

SP - 267

EP - 276

BT - Methods in Molecular Biology

PB - Humana Press Inc.

ER -