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Carcinogenesis Advance Access published online on July 20, 2005
Carcinogenesis, doi:10.1093/carcin/bgi184
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© The Author 2005. Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oupjournals.org
Received April 29, 2005
Revised July 10, 2005
Accepted July 12, 2005

CANCER BIOLOGY

Screening for genomic fragments that are methylated specifically in colorectal carcinoma with a methylated MLH1 promoter

Koji Koinuma 1, Ruri Kaneda 2, Minoru Toyota 3, Yoshihiro Yamashita 2, Shuji Takada 2, Young Lim Choi 2, Tomoaki Wada 2, Masaki Okada 4, Fumio Konishi 5, Hideo Nagai 4, and Hiroyuki Mano 6*

1 Division of Functional Genomics, Jichi Medical School, Tochigi 329-0498, Japan; Department of Surgery, Jichi Medical School, Tochigi 329-0498, Japan
2 Division of Functional Genomics, Jichi Medical School, Tochigi 329-0498, Japan
3 Department of Molecular Biology, Cancer Research Institute, Sapporo Medical University, Hokkaido 060-8556, Japan
4 Department of Surgery, Jichi Medical School, Tochigi 329-0498, Japan
5 Department of Surgery, Omiya Medical Center of Jichi Medical School, Saitama 330-8503, Japan
6 Division of Functional Genomics, Jichi Medical School, Tochigi 329-0498, Japan; CREST, Japan Science and Technology Agency, Saitama 332-0012, Japan

* To whom correspondence should be addressed.
Hiroyuki Mano, E-mail: hmano{at}jichi.ac.jp


  Abstract

A subset of colorectal carcinomas (CRCs) is associated with microsatellite instability (MSI) of the genome. Although extensive methylation of CpG islands within the promoter regions of DNA mismatch repair genes such as MLH1 is thought to play a central role in tumorigenesis for MSI-positive sporadic CRCs, it has been obscure whether such aberrant epigenetic regulation occurs more widely and affects other cancer-related genes in vivo. Here, by using the methylated CpG island amplification coupled with representational difference analysis (MCA-RDA), we screened genomic fragments that are selectively methylated in CRCs positive for MLH1 methylation, resulting in the identification of hundreds of CpG island-containing genomic fragments. Methylation status of such CpG islands was verified for 28 genomic clones in eight CRC specimens positive for MLH1 methylation and the corresponding paired normal colon tissue as well as in eight CRC specimens negative for MLH1 methylation. Many of the CpG islands were preferentially methylated in the MLH1 methylation-positive CRC specimens, although methylation of some of them was more widespread. These data provide insight into the complex regulation of the methylation status of CpG islands in CRCs positive for MSI and MLH1 methylation.


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