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The Retinoblastoma Protein pp 35–47Cite as

Detection of Aberrant DNA Methylation Patterns in the RB1 Gene

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1726))

Abstract

The retinoblastoma protein (pRb) plays a central role in the regulation of cell cycle by interaction with members of the E2F transcription factor family. As a tumor suppressor protein, pRb is frequently dysregulated in several major cancers. In addition to mutations, inactivation of pRb is also caused by epigenetic mechanisms including alterations of DNA methylation. There are three CpG islands located within the RB1 gene that encodes pRb that are closely associated with the regulation of pRb expression. Aberrant DNA methylation at the RB1 gene has been reported in sporadic retinoblastoma as well as other cancers including glioblastoma, hepatocellular carcinoma, and breast cancer. Recent studies have revealed that the RB1 gene is imprinted. Therefore, quantitative analysis is required to detect aberrations in DNA methylation associated with imprint deregulation. Pyrosequencing® is considered as the method of choice for quantitative and reproducible analysis of DNA methylation with single base resolution. In this chapter, we provide a detailed protocol for the quantitative analysis of RB1 gene methylation using bisulfite Pyrosequencing®.

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Authors and Affiliations

  1. Department of Surgery, Faculty of Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia

    Sumadi Lukman Anwar

  2. Institute of Pathology, Medizinische Hochschule Hannover, Hannover, Germany

    Ulrich Lehmann

Authors
  1. Sumadi Lukman Anwar
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  2. Ulrich Lehmann
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Corresponding author

Correspondence to Ulrich Lehmann .

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Editors and Affiliations

  1. Biochemistry and Cancer Biology Divisions Basic Science Department, Ponce Health Sciences University, Ponce, Puerto Rico

    Pedro G. Santiago-Cardona

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Anwar, S.L., Lehmann, U. (2018). Detection of Aberrant DNA Methylation Patterns in the RB1 Gene. In: Santiago-Cardona, P. (eds) The Retinoblastoma Protein. Methods in Molecular Biology, vol 1726. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7565-5_5

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  • DOI: https://doi.org/10.1007/978-1-4939-7565-5_5

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7564-8

  • Online ISBN: 978-1-4939-7565-5

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