Methylation-specific ligation detection reaction (msLDR): A new approach for multiplex evaluation of methylation patterns

Publikation: Beitrag in FachzeitschriftForschungsartikelBeigetragenBegutachtung

Beitragende

  • Felix Bormann - , Charité – Universitätsmedizin Berlin (Autor:in)
  • Christine Sers - , Charité – Universitätsmedizin Berlin (Autor:in)
  • Barbara Seliger - , Martin-Luther-Universität Halle-Wittenberg (Autor:in)
  • Diana Handke - , Martin-Luther-Universität Halle-Wittenberg (Autor:in)
  • Thomas Bergmann - , Max Planck Institut für Molekulare Genetik (Autor:in)
  • Stephanie Seibt - , Charité – Universitätsmedizin Berlin (Autor:in)
  • Hans Lehrach - , Max Planck Institut für Molekulare Genetik (Autor:in)
  • Andreas Dahl - , DRESDEN-concept Genome Center (CMCB Core Facility) (Autor:in)

Abstract

A new sensitive method for multiplex gene-specific methylation analysis was developed using a ligation-based approach combined with a TaqMan-based detection and readout employing universal reporter probes. The approach, termed methylation-specific Ligation Detection Reaction (msLDR), was applied to test 16 loci in 8 different colorectal cancer cells in parallel. These loci encode immune regulatory genes involved in T-cell and natural killer cell activation, whose silencing is associated with the development or progression of colorectal cancer. Parallel analysis of HLA-A, HLA-B, STAT1, B2M, LMP2, LMP7, PA28α, TAP1, TAP2, TAPBP, ULBP2 and ULBP3 by msLDR in eight colorectal cancer cell lines showed preferential methylation at the HLA-B, ULBP2 and ULBB3 loci, but not at the other loci. MsLDR was found to represent a suitable and sensitive method for the detection of distinct methylation patterns as validated by conventional bisulphite Sanger sequencing and COBRA analysis. Since gene silencing by epigenetic mechanisms plays a central role during transformation of a normal differentiated somatic cell into a cancer cell, characterization of the gene methylation status in tumours is a major topic not only in basic research, but also in clinical diagnostics. Due to a very simple workflow, msLDR is likely to be applicable to clinical samples and thus comprises a potential diagnostic tool for clinical purposes.

Details

OriginalspracheEnglisch
Seiten (von - bis)279-291
Seitenumfang13
FachzeitschriftMolecular genetics and genomics
Jahrgang286
Ausgabenummer3-4
PublikationsstatusVeröffentlicht - Okt. 2011
Peer-Review-StatusJa

Externe IDs

PubMed 21879293

Schlagworte

ASJC Scopus Sachgebiete

Schlagwörter

  • Cancer, DNA-methylation, Epigenetics, Immune genes, Multiplex