Heterogeneity of DNA methylation status analyzed by bisulfite-PCR-SSCP andcorrelation with clinico-pathological characteristics in colorectal cancer

Citation
M. Maekawa et al., Heterogeneity of DNA methylation status analyzed by bisulfite-PCR-SSCP andcorrelation with clinico-pathological characteristics in colorectal cancer, CLIN CH L M, 39(2), 2001, pp. 121-128
Citations number
29
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Medical Research Diagnosis & Treatment
Journal title
CLINICAL CHEMISTRY AND LABORATORY MEDICINE
ISSN journal
1434-6621 → ACNP
Volume
39
Issue
2
Year of publication
2001
Pages
121 - 128
Database
ISI
SICI code
1434-6621(200102)39:2<121:HODMSA>2.0.ZU;2-0
Abstract
Aberrant DNA methylation has been identified as an important mechanism for inactivation of tumor suppressor genes and mismatch repair genes during car cinogenesis. We used bisulfite treatment and the PCR-single strand conforma tion polymorphism (SSCP) (BiPS) technique to analyze methylation status of the promoter regions of the hMLH1, p16, and HIC1 genes in several cancer ce ll 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 h omogeneity 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 reve aled 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 fre quently and could serve as a sensitive marker for colorectal cancer. Methyl ation status of the hMLH1 and p16 gene promoters was correlated with micros atellite instability status, tumor location, and differentiation but not wi th K-ras mutation or allelic loss of p53.