Cloning of the hamster p16 gene 5' upstream region and its aberrant methylation patterns in pancreatic cancer.
The hamster model of pancreatic carcinogenesis is useful for understanding the development of human pancreatic cancer. However, there is only a small amount of hamster genetic information available for analyzing the gene alterations in hamster pancreatic cancers. Here, we determined the nucleotide sequence of the 5' upstream region of the hamster p16 gene using a suppression polymerase chain reaction method combined with gene-specific primers. Based on this sequence, we analyzed the methylation status of the 5' region by bisulfite sequencing in three normal pancreatic tissues and five pancreatic duct adenocarcinomas (PDAs). All five PDAs were highly methylated in the 5' upstream region and showed reduced expressions of the p16 gene, while the three normal samples were demethylated. The method described in this study is a highly effective and rapid technique for determining the 5' upstream region, and is applicable to epigenetic studies of the methylation status of this region.
Laboratory of Cancer Biology and Bioinformatics, Department of Life Science, Faculty of Science and Engineering, Kinki University, 3-4-1, Kowakae, Higashiosaka, Osaka 577-8502, Japan.
Madoka
M
Hanaoka
Kyoko
K
Shimizu
Mayumi
M
Shigemura
Ayumi
A
Kato
Hiromasa
H
Fujii
Kanya
K
Honoki
Toshifumi
T
Tsujiuchi
D024506:5' Flanking Region / Q000235:genetics*; D000818:Animals; D002869:Chromosome Aberrations; D003001:Cloning, Molecular / Q000379:methods*; D006224:Cricetinae; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000235:genetics*; D019175:DNA Methylation; D005260:Female; D019942:Genes, p16; D006801:Humans; D008297:Male; D008647:Mesocricetus; D010190:Pancreatic Neoplasms / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods; D011401:Promoter Regions, Genetic; D017422:Sequence Analysis, DNA / Q000379:methods
D019941:Cyclin-Dependent Kinase Inhibitor p16