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Japanese Journal of Clinical Oncology Advance Access originally published online on March 19, 2008
Japanese Journal of Clinical Oncology 2008 38(4):317-322; doi:10.1093/jjco/hyn021
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© The Author (2008). Published by Oxford University Press. All rights reserved

Fluorescence-labeled Methylation-sensitive Amplified Fragment Length Polymorphism (FL-MS-AFLP) Analysis for Quantitative Determination of DNA Methylation and Demethylation Status

Shinji Kageyama1,2, Kazuya Shinmura1, Hiroko Yamamoto1,2, Masanori Goto1, Koichi Suzuki3, Fumihiko Tanioka1,4, Toshihiro Tsuneyoshi2 and Haruhiko Sugimura1

1 1st Department of Pathology, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka
2 Department of Materials and Life Science, Shizuoka Institute of Science and Technology, Fukuroi, Shizuoka
3 Department of Surgery, Omiya Medical Center, Jichi Medical School, Saitama
4 Department of Pathology, Iwata City Hospital, Iwata, Shizuoka, Japan

For reprints and all correspondence: Haruhiko Sugimura, 1st Department of Pathology, Hamamatsu University School of Medicine, 1-20-1 Handayama, Higashi Ward, Hamamatsu, Shizuoka 431-3192, Japan. E-mail: hsugimur{at}hama-med.ac.jp

Received November 27, 2007; accepted February 16, 2008

The PCR-based DNA fingerprinting method called the methylation-sensitive amplified fragment length polymorphism (MS-AFLP) analysis is used for genome-wide scanning of methylation status. In this study, we developed a method of fluorescence-labeled MS-AFLP (FL-MS-AFLP) analysis by applying a fluorescence-labeled primer and fluorescence-detecting electrophoresis apparatus to the existing method of MS-AFLP analysis. The FL-MS-AFLP analysis enables quantitative evaluation of more than 350 random CpG loci per run. It was shown to allow evaluation of the differences in methylation level of blood DNA of gastric cancer patients and evaluation of hypermethylation and hypomethylation in DNA from gastric cancer tissue in comparison with adjacent non-cancerous tissue.

Key Words: DNA fingerprinting • FL-MS-AFLP • gastric cancer • hypermethylation • hypomethylation


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