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Epigenetic typing of human cancer cell lines by BlsI- and GlaI-PCR assays

This email address is being protected from spambots. You need JavaScript enabled to view it. , Danila A. Gonchar, Murat A. Abdurashitov and Sergey Kh. Degtyarev

Translated from "Ovchinnikov bulletin of biotechnology and physical and chemical biology" V.7, No 3, pp 5-16, 2011

 

BlsI- and GlaI-PCR assays have been applied to determine a methylation status of regulatory regions of SEPT9b, IGFBP3, CEBPD, MGMT and RASSF1A tumor suppressor genes in malignant cell lines HeLa, Raji, U-937, Jurkat and in the control fibroblast cell line L-68. GlaI- and BlsI-PCR assays have shown either presence or absence of 5’-R(5mC)GY-3’ sites in the regulatory regions of the studied tumor suppressor genes depending on malignant cell line, which was a source of DNA preparation. Regulatory regions are methylated in different combinations in the studied malignant cell lines. At the same time no 5’-R(5mC)GY-3’ sites have been found in these regulatory regions in DNA from normal fibroblast cell line L-68. These results show that method of BlsI- and GlaI-PCR assays allow to determine and to discriminate malignant cell lines. Thus, BlsI- and GlaI-PCR assays may be used for epigenetic typing of malignant cell lines.

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BlsI- and GlaI-PCR assays – a new method of DNA methylation study

This email address is being protected from spambots. You need JavaScript enabled to view it. , A. G. Akishev, S. Kh. Degtyarev

 

 

BlsI- and GlaI-PCR assays have been developed to study DNA methylation. A new method includes DNA hydrolysis by methyl-directed site-specific DNA endonucleases GlaI or BlsI with subsequent routine PCR. Study of DNA methylation in regulation region of human tumor suppressor genes has been performed for a new method evaluation. BlsI- and GlaI-PCR assays have revealed different methylation patterns in promoter region of DAPK1, in promoter and first exon region of RARB and in first exon region of RASSF1A tumor suppressor genes in malignant cell lines HeLa, Raji, U-937, Jurkat and control L-68 cells. GlaI-PCR assay has shown methylation of RARB promoter and first exon region in DNA from all malignant cell lines, but not in control L-68 cells. GlaI- and BlsI-PCR assays have displayed DNA methylation of RASSF1A first exon region in Raji and Jurkat cells only. BlsI-PCR assay of DAPK1 promoter region has demonstrated an additional DNA methylation in Raji cells only. GlaI- and BlsI-PCR assays may be useful both in determination of malignant cells and their discrimination. 

Read more: BlsI- and GlaI-PCR assays – a new method of DNA methylation study