Carcinogenesis Advance Access originally published online on July 20, 2005
Carcinogenesis 2005 26(12):2078-2085; doi:10.1093/carcin/bgi184
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Carcinogenesis vol.26 no.12 © Oxford University Press 2005; all rights reserved.
Screening for genomic fragments that are methylated specifically in colorectal carcinoma with a methylated MLH1 promoter
1 Division of Functional Genomics and 2 Department of Surgery, 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, Omiya Medical Center of Jichi Medical School, Saitama 330-8503, Japan and 5 CREST, Japan Science and Technology Agency, Saitama 332-0012, Japan
* To whom correspondence should be addressed. Tel: +81 285 58 7449; Fax: +81 285 44 7322; E-mail: hmano{at}jichi.ac.jp
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 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 islands containing genomic fragments. Methylation status of such CpG islands was verified for 28 genomic clones in 8 CRC specimens positive for MLH1 methylation and the corresponding paired normal colon tissue as well as in 8 CRC specimens negative for 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 insights into the complex regulation of the methylation status of CpG islands in CRCs positive for MSI and MLH1 methylation.
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