Malyshev B S, Netesova N A, Smetannikova N A, Abdurashitov M A, Akishev A G, Dubinin E V, Azanov A Z, Vihlyanov I V, Nikitin M K, Karpov A B, Degtyarev S Kh
State Research Center of Virology and Biotechnology «Vector», Novosibirsk region, Koltsovo, 630559 Russia.
EpiGene LLC, Novosibirsk, 630090 Russia.
Acta Naturae. 2020 Jul-Sep;12(3):124-133. doi: 10.32607/actanaturae.11070.
At early stages of carcinogenesis, the regulatory regions of some tumor suppressor genes become aberrantly methylated at RCGY sites, which are substrates of DNA methyltransferase Dnmt3. Identification of aberrantly methylated sites in tumor DNA is considered to be the first step in the development of epigenetic PCR test systems for early diagnosis of cancer. Recently, we have developed a GLAD-PCR assay, a method for detecting the R(5mC)GY site in the genome position of interest even at significant excess of DNA molecules with a non-methylated RCGY site in this location. The aim of the present work is to use the GLAD-PCR assay to detect the aberrantly methylated R(5mC)GY sites in the regulatory regions of tumor suppressor genes (, , , , , , , , , , , , , , , , , , , , , , and ) in DNA samples obtained from gastric cancer (GC) tissues. The study of the DNA samples derived from 29 tumor and 25 normal gastric tissue samples demonstrated a high diagnostic potential of the selected RCGY sites in the regulatory regions of the , and genes; the total indices of sensitivity and specificity for GC detection being 96.6% and 100%, respectively.
在致癌作用的早期阶段，一些肿瘤抑制基因的调控区域在RCGY位点发生异常甲基化，这些位点是DNA甲基转移酶Dnmt3的作用底物。鉴定肿瘤DNA中的异常甲基化位点被认为是开发用于癌症早期诊断的表观遗传PCR检测系统的第一步。最近，我们开发了一种GLAD-PCR检测方法，该方法即使在感兴趣的基因组位置存在大量未甲基化RCGY位点的DNA分子时，也能检测该位置的R(5mC)GY位点。本研究的目的是使用GLAD-PCR检测方法，检测从胃癌（GC）组织获得的DNA样本中肿瘤抑制基因（、、、、、、、、、、、、、、、、、、、、、和）调控区域中异常甲基化的R(5mC)GY位点。对来自29个肿瘤和25个正常胃组织样本的DNA样本进行的研究表明，、和基因调控区域中选定的RCGY位点具有很高的诊断潜力；GC检测的敏感性和特异性总指标分别为96.6%和100%。
