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

Bormann, F., Sers, C., Seliger, B., Handke, D., Bergmann, T., Seibt, S., et al. (2011). Methylation-specific ligation detection reaction (msLDR): a new approach for multiplex evaluation of methylation patterns. Molecular Genetics and Genomics: MGG, 286(3-4), 279-91. Retrieved from http://www.ncbi.nlm.nih.gov/pubmed/21879293 http://www.springerlink.com/content/eq76180447161h22/fulltext.pdf.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-78C8-E Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-78C9-C
Genre: Journal Article

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Bormann, F., Author
Sers, C., Author
Seliger, B., Author
Handke, D., Author
Bergmann, T.1, Author              
Seibt, S., Author
Lehrach, H.1, Author              
Dahl, A.1, Author              
Affiliations:
1Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1433550              

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Free keywords: Antigen Presentation/genetics; Cell Line, Tumor; Colorectal Neoplasms/chemistry/genetics/immunology; *DNA Methylation; DNA, Neoplasm/chemistry/genetics; GPI-Linked Proteins/genetics; Genes, MHC Class I; *Genetic Techniques/statistics & numerical data; Humans; Intercellular Signaling Peptides and Proteins/genetics; Killer Cells, Natural/immunology; Miniaturization; Polymerase Chain Reaction/methods; T-Lymphocytes/immunology
 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, PA28alpha, 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.

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 Dates: 2011
 Publication Status: Published in print
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Title: Molecular Genetics and Genomics : MGG
Source Genre: Journal
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Pages: - Volume / Issue: 286 (3-4) Sequence Number: - Start / End Page: 279 - 91 Identifier: ISSN: 1617-4623 (Electronic) 1617-4623 (Linking)