alexa GLAD-PCR Assay of DNA Methylation Markers Associated with Colorectal Cancer
ISSN: 0974-8369

Biology and Medicine
Open Access

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Research Article

GLAD-PCR Assay of DNA Methylation Markers Associated with Colorectal Cancer

Alexey A Evdokimov1, Nina A Netesova1, Natalia A Smetannikova1, Murat A Abdurashitov1*, Alexandr G Akishev1, Boris S Malyshev1, Evgeniya S Davidovich1, Vladimir V Fedotov1, Vitaliy V Kuznetsov1, Yuriy D Ermolaev2, Andrey B Karpov2, Alexey E Sazonov2, Ravil M Tahauov2 and Sergey Kh Degtyarev1

1State Research Center of Virology and Biotechnology “Vector,” Novosibirsk 630559, Russia

2Seversk Biophysical Research Centre of the Federal Medico-Biological Agency, Seversk 636000, Russia

*Corresponding Author:
Abdurashitov MA
SibEnzyme, 2/12 Ak.Timakov Str.
Novosibirsk 630117, Russia
Tel: +7 383 3336854
Fax: +7 383 3336853
E-mail: [email protected]

Received Date: August 04, 2016; Accepted Date: September 01, 2016; Published Date: September 08, 2016

Citation: Evdokimov AA, Netesova NA, Smetannikova NA, Abdurashitov MA, Akishev AG, et al. (2016) GLAD-PCR Assay of DNA Methylation Markers Associated with Colorectal Cancer. Biol Med (Aligarh) 8: 342. doi:10.4172/0974-8369.1000342

Copyright: © 2016 Evdokimov et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.



Hypermethylation of the gene regulatory regions is documented for many cancer diseases. Such an aberrant DNA methylation in cancer cells is catalyzed by DNA methyltransferases Dnmt3a and Dnmt3b, which predominantly recognize and methylate RCGY sequences with formation of R(5mC)GY sites. Recently, based on a new methyl-directed DNA endonuclease GlaI, we developed a GLAD-PCR assay, which allows determining R(5mC)GY site in a defi ned position of the genomic DNA. In this work we applied GLAD-PCR assay for identifi cation of the methylated RCGY sites in the regulatory regions of some downregulated genes associated with colorectal cancer (CRC). This list includes ADHFE1, ALX4, CNRIP1, EID3, ELMO1, ESR1, FBN1, HLTF, LAMA1, NEUROG1, NGFR, RARB, RXRG, RYR2, SDC2, SEPT9, SFRP2, SOCS3, SOX17, THBD, TMEFF2, UCHL1, and VIM genes. GLAD-PCR analysis of selected RCGY sites within the regulatory regions of some of these genes demonstrates a good prognostic potential with relatively high sensitivity and specifi city of CRC detection in tumor DNA.


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