Masato Maekawa, Kokichi Sugano, Mineko Ushiama, Noriko Fukayama, Kiyoaki Nomoto, Hidefumi Kashiwabara, Shin Fujita, Tadao Kakizoe
Heterogeneity of DNA Methylation Status Analyzed by Bisulfite-PCR-SSCP and Correlation with Clinico-Pathological Characteristics in Colorectal Cancer
Aberrant DNA methylation has been identified as an
important mechanism for inactivation of tumor suppressor
genes and mismatch repair genes during carcinogenesis.
We used b?i?sulfite treatment and the P?CR s?ingle
strand conformation polymorphism (S?SCP)
(BiPS) technique to analyze methylation status of the
promoter regions of the hMLH1, p16, and HIC1 genes in
several cancer cell lines and colorectal cancer tissues.
The methylation of the hMLH1, p16 and HIC1 genes
was observed in 2, 8, and 13 of 13 cancer cell lines, respectively.
The SSCP for p16 and HIC1 in each of the
methylation-positive cell lines were similar, indicating
relative homogeneity of methylation status and complete
methylation in the cell lines. Methylation was observed
in 8, 5, and 21 of 25 colorectal cancer tissues for
the hMLH1, p16, and HIC1 genes, respectively. The
methylated bands revealed by BiPS analysis of the
hMLH1 gene were homogeneous, whereas those of
the p16 and HIC1 genes were different in each case.
The methylation of the promoter region of the HIC1
gene in colorectal cancer was observed most frequently
and could serve as a sensitive marker for colorectal
cancer. Methylation status of the hMLH1 and
p16 gene promoters was correlated with microsatellite
instability status, tumor location, and differentiation
but not with K-ras mutation or allelic loss of p53.
Clinical Chemical Laboratory Medicine, Walter de Gruyter
Print ISSN: 1434-6621
Volume: 39, 04/2001
Pages: 121 - 128
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