Methylation profiles of genes utilizing newly developed CpG island methylation microarray on colorectal cancer patients.
Aberrant methylation of DNA has been shown to play an important role in a variety of human cancers, developmental disorders and aging. Hence, aberrant methylation patterns in genes can be a molecular marker for such conditions. Therefore, a reliable but uncomplicated method to detect DNA methylation is preferred, not merely for research purposes but for daily clinical practice. To achieve these aims, we have established a precise system to identify DNA methylation patterns based on an oligonucleotide microarray technology. Our microarray method has an advantage over conventional methods and is unique because it allows the precise measurement of the methylation patterns within a target region. Our simple signal detection system depends on using an avidin-biotinylated peroxidase complex and does not require an expensive laser scanner or hazardous radioisotope. In this study, we applied our technique to detect promoter methylation status of O6-methylguanine-DNA methyltransferase (MGMT) gene. Our easy-handling technology provided reproducible and precise measurement of methylated CpGs in MGMT promoter and, thus, our method may bring about a potential evolution in the handling of a variety of high-throughput DNA methylation analyses for clinical purposes.
Kimura
Naoki
N
Research and Development Center, Nisshinbo Industries Inc, 1-2-3 Ohnodai, Midori-ku, Chiba-shi, Chiba 267-0056, Japan.
Nagasaka
Takeshi
T
Murakami
Jun
J
Sasamoto
Hiromi
H
Murakami
Masahiro
M
Tanaka
Noriaki
N
Matsubara
Nagahide
N
D001360:Avidin / Q000031:analogs & derivatives*; D015179:Colorectal Neoplasms / Q000235:genetics* / Q000378:metabolism; D018899:CpG Islands; D019175:DNA Methylation; D006735:Horseradish Peroxidase; D006801:Humans; D019853:O(6)-Methylguanine-DNA Methyltransferase / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic
C083986:avidin-horseradish peroxidase complex; D001360:Avidin; D006735:Horseradish Peroxidase; D019853:O(6)-Methylguanine-DNA Methyltransferase