Hanaoka Madoka, Shimizu Kyoko, Shigemura Mayumi, Kato Ayumi, Fujii Hiromasa, Honoki Kanya, Tsujiuchi Toshifumi
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.
Biochem Biophys Res Commun. 2005 Aug 12;333(4):1249-53. doi: 10.1016/j.bbrc.2005.06.034.
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.
仓鼠胰腺癌发生模型对于理解人类胰腺癌的发展很有用。然而，可用于分析仓鼠胰腺癌基因改变的仓鼠遗传信息很少。在此，我们使用抑制聚合酶链反应方法结合基因特异性引物，确定了仓鼠p16基因5'上游区域的核苷酸序列。基于该序列，我们通过亚硫酸氢盐测序分析了三个正常胰腺组织和五个胰腺导管腺癌（PDA）中5'区域的甲基化状态。所有五个PDA在5'上游区域高度甲基化，p16基因表达降低，而三个正常样本去甲基化。本研究中描述的方法是确定5'上游区域的高效快速技术，适用于该区域甲基化状态的表观遗传学研究。
