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Analysis of DNA methylation of isolated glands in gastric cancers and the surrounding the intestinal glands and non -intestinal glands

胃癌离体腺体及其周围肠腺和非肠腺DNA甲基化分析

基本信息

批准号：
22590317
负责人：
SUGAI Tamotsu
金额：
$ 3万
依托单位：
Iwate Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2010
资助国家：
日本
起止时间：
2010 至 2012
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-22590317/
关键词：
胃腸上皮化生メチル化腺管分離 Epigenetics

项目摘要

The crypt isolation technique was used to analysis methylation of tumor related genes of non-metaplastic g astric gland (NM), intestinal metaplasia gland (IM) and gastric cancer gland (GC). NM, IM and GC specimens were obtained from 35 patients with gastric cancer (intestinal type [n=22], diffuse type [n=13]). Three samples were obtained from cancer, antrum and body. After crypt isolation of each sample, the non-cancerous glands were classified into NM glands, incomplete intestinal metaplasia gland (IIM) and complete intestinal metaplasia gland (CIM) by double stain methods of alcian blue and MUC5AC immnostain. These samples were examined CpG island hypermethylation by Combine Bislufite Restriction Analysis (COBRA) for 8 genes (MLH1 , p16 , HPP1, RUNX3 , ZFP64 , RASSF2A, SFRP1, E-cadherin). IIM and CIM gland have significant higher methylation than NM gland. But, IIM and CIM had no significant differences in methylation status. The methylation GC gland and diffuse type GC gland had no significant differences. The methylation level of GC gland was significantly higher than corresponding NM gland, but no difference was observed in corresponding IM gland. The present data on NM gland, GIM gland and GC gland suggest that genetic alternations already occur within the surrounding noncancerous glands.status of intestinal type .

采用腺泡分离技术检测非化生胃腺（NM）、肠化生胃腺（IM）和胃癌胃腺（GC）肿瘤相关基因甲基化。35例胃癌患者（肠型22例，弥漫型13例）的NM、IM和GC标本。从癌、胃窦和身体获得三个样品。每例标本经腺泡分离后，用阿辛蓝和MUC 5AC免疫组化双重染色法将非癌腺体分为NM腺、不完全肠化腺（IIM）和完全肠化腺（CIM）。采用联合收割机双硫酶限制性分析（COBRA）检测MLH 1、p16、HPP 1、RUNX 3、ZFP 64、RASSF 2A、SFRP 1、E-cadherin等8个基因的CpG岛甲基化。IIM和CIM腺甲基化水平显著高于NM腺。但IIM和CIM的甲基化状态无显著差异。甲基化型胃癌腺体与弥漫型胃癌腺体无显著性差异。GC腺甲基化水平显著高于相应NM腺，而相应IM腺甲基化水平无显著差异。NM腺、GIM腺和GC腺的现有资料表明，在其周围的非癌腺体中已经发生了遗传变异。