The application of methylation specific electrophoresis (MSE) to DNA methylation analysis of the 5' CpG island of mucin in cancer cells.

BMC Cancer
Seiya YokoyamaSuguru Yonezawa

Abstract

Methylation of CpG sites in genomic DNA plays an important role in gene regulation and especially in gene silencing. We have reported mechanisms of epigenetic regulation for expression of mucins, which are markers of malignancy potential and early detection of human neoplasms. Epigenetic changes in promoter regions appear to be the first step in expression of mucins. Thus, detection of promoter methylation status is important for early diagnosis of cancer, monitoring of tumor behavior, and evaluating the response of tumors to targeted therapy. However, conventional analytical methods for DNA methylation require a large amount of DNA and have low sensitivity. Here, we report a modified version of the bisulfite-DGGE (denaturing gradient gel electrophoresis) using a nested PCR approach. We designated this method as methylation specific electrophoresis (MSE). The MSE method is comprised of the following steps: (a) bisulfite treatment of genomic DNA, (b) amplification of the target DNA by a nested PCR approach and (c) applying to DGGE. To examine whether the MSE method is able to analyze DNA methylation of mucin genes in various samples, we apply it to DNA obtained from state cell lines, ethanol-fixed colonic crypts and human pancre...Continue Reading

References

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Citations

Jun 21, 2013·Expert Review of Molecular Diagnostics·Jesse HintonJinping Li
Jan 12, 2016·Biology·Sergey Kurdyukov, Martyn Bullock
Feb 5, 2014·Biochemical and Biophysical Research Communications·Wei GuangErik P Lillehoj
Jul 7, 2017·Genes & Cancer·Seiya YokoyamaAkihide Tanimoto
Jan 30, 2020·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Seiya YokoyamaAkihide Tanimoto
Apr 8, 2021·BioMed Research International·Ehsan KhodadadiHossein Samadi Kafil

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Datasets Mentioned

BETA
MDA-MB-453

Methods Mentioned

BETA
PCR
electrophoresis
Assay

Software Mentioned

MassARRAY
Image J
methylight

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