Evdokimov A A, Netesova N A, Smetannikova N A, Abdurashitov M A, Akishev A G, Davydovich E S, Ermolaev Yu D, Karpov A B, Sazonov A E, Takhauov R M, Degtyarev S Kh
Vopr Onkol. 2016;62(1):117-21.
Aberrant methylation of regulation regions of tumorsuppressor genes is showed for many cancer diseases. In course of this modification an enzyme DNMT3 methylates RCGY sites in CpG-islands of regulation regions producing R(5mC)GY sites. Earlier we developed GLAD-PCR assay to determine R(5mC)GY site in a definite position of human genome. In this work we have applied GLAD-PCR assay to determine R(5mC)GY sites in regulation regions of ESR1 and ELMO1 tumor-suppressor genes. We have studied a fragment of first exon of ELMO1 gene and a part of ESR1 promoter region in DNA preparations from malignant cell line SW837 and colorectal tumor samples. We have checked four sites in each region and found two highly methylated sites: GCGC in first exon of ELMO1 gene and GCGT in promoter region of ESR1 gene. Site GCGT is weakly methylated in healthy tissues and more methylated in the most of colorectal samples. Site GCGC is not methylated in healthy tissues and significantly methylated in 60% of colorectal samples. A possibility to use GLAD-PCR assay for cancer diagnostics is discussed.
许多癌症疾病都表现出肿瘤抑制基因调控区域的异常甲基化。在这种修饰过程中，一种名为DNMT3的酶会使调控区域CpG岛中的RCGY位点甲基化，产生R(5mC)GY位点。此前我们开发了GLAD-PCR检测法来确定人类基因组特定位置的R(5mC)GY位点。在这项工作中，我们应用GLAD-PCR检测法来确定ESR1和ELMO1肿瘤抑制基因调控区域中的R(5mC)GY位点。我们研究了恶性细胞系SW837和结直肠肿瘤样本DNA制剂中ELMO1基因第一外显子的一个片段和ESR1启动子区域的一部分。我们在每个区域检测了四个位点，发现了两个高度甲基化的位点：ELMO1基因第一外显子中的GCGC和ESR1基因启动子区域中的GCGT。GCGT位点在健康组织中甲基化程度较低，而在大多数结直肠样本中甲基化程度较高。GCGC位点在健康组织中未甲基化，而在60%的结直肠样本中显著甲基化。本文还讨论了使用GLAD-PCR检测法进行癌症诊断的可能性。
