title: Allele-specific methylation analysis on upstream promoter region of H19 by methylation-specific PCR with confronting two-pair primers.
abstract: H19 and IGF2 genes are imprinted genes and expressed differently depending on whether they are carried by a chromosome of maternal or paternal origin; H19 is expressed only from the maternal allele and IGF2 only from the paternally inherited allele. The upstream promoter region of H19 has the imprinting-control region (ICR) or CTCF binding sites, where the methylation status of this region is critical to the regulation of imprinting of the H19/IGF2 locus located in chromosome 11p15. There are various reports on imprinting disorders in this region. In colorectal cancer aberrant biallelic methylation of CTCF binding site has been reported, and aberrant hypomethylation of this region in bladder cancer. Thus, certain human neoplasms have either hyper- or hypo-methylation in the ICR. Hence it is still difficult to analyze allele-specific methylation disorder of the region, or differentially methylated regions (DMR), locate upstream of H19. Here we report a new method, which could distinguish paternal epigenetic or maternal epigenetic pattern by a single PCR assay, to combine methylation-specific PCR and PCR with confronting two-pair primers (MSP-CTPP). Using this method, we investigated the region close to H19 ICR in 161 colorectal cancer and 65 gastric cancer cases.
authors.affiliation: Department of Gastroenterological Surgery and Surgical Oncology, Okayama University Graduate School of Medicine and Dentistry, 2-5-1 Shikata-cho, Okayama 700-8558, Japan.
authors.forename: Hiromi
authors.initials: H
authors.lastname: Sasamoto
authors.forename: Takeshi
authors.initials: T
authors.lastname: Nagasaka
authors.forename: Kenji
authors.initials: K
authors.lastname: Notohara
authors.forename: Kazuhide
authors.initials: K
authors.lastname: Ozaki
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Isozaki
authors.forename: Noriaki
authors.initials: N
authors.lastname: Tanaka
authors.forename: Nagahide
authors.initials: N
authors.lastname: Matsubara
mesh_terms: D000368:Aged; D000483:Alleles; D015179:Colorectal Neoplasms / Q000235:genetics*; D019175:DNA Methylation; D017931:DNA Primers; D005260:Female; D018392:Genomic Imprinting; D006801:Humans; D007413:Intestinal Mucosa; D008297:Male; D018895:Microsatellite Repeats; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D011401:Promoter Regions, Genetic; D062085:RNA, Long Noncoding; D022661:RNA, Untranslated / Q000235:genetics* / Q000378:metabolism*; D013274:Stomach Neoplasms / Q000235:genetics*
chemical_list: D017931:DNA Primers; C094335:H19 long non-coding RNA; D062085:RNA, Long Noncoding; D022661:RNA, Untranslated