Analysis of a new gene alteration related to fatty acid synthase in colorectal carcinogenesis.

结直肠癌发生中与脂肪酸合酶相关的新基因改变的分析。

• 批准号: 13671337
• 负责人: HASEGAWA Hirotoshi
• 金额: $ 2.37万
• 依托单位: Keio University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671337/
• 关键词: colorectal cancer; Restriction landmark genomic scanning; gene mutation; fatty acid; Human Not I clones

[Background] We identified a new genetic alteration in human hepatocellular carcinoma by using a high resolution technique, restriction landmark genomic scanning (RLGS). The aim of this study was to identify new genetic alterations in colorectal cancer using the same method.[Methods] High molecular weight genomic DNAs were extracted from 25 colorectal cancerous tissues and corresponding normal mucosae. Each DNA was cleaved with the restriction enzyme Not I, size fractionated by 1st-dimensional electrophoresis using Pvu II, and the DNA fragments were then cleaved by Pst I as the third enzyme in gel and separated by 2-dimensional electrophoresis. By comparing each pair of RLGS profiles, we detected 6 common changed spots. The most frequently changed spot was directly cloned from the gel. Nineteen of 25 (76.0%) colorectal cancer cases showed a spot with the same alteration, which never appeared in normal mucosae. A 410-bp DNA fragment corresponding to this spot was cloned. This sequence was found to be 69% homologous to part of the human fatty acid synthase gene (FAS), and mapped to 95% of the 3rd chromosome of the human draft sequence. The three cloned spots (Spots B, C and F) were observed in 44%(11/25), 24% (6/25) and 28% (7/25) of the cases, respectively, and found to be 271-, 168-, and 374-bp DNA fragments. The DNA sequences from spots B, C and F were also homologous to the genomic contig of the first and the ninth chromosomes, and human Not I clones, respectively.[Conclusion] Four spots were cloned, one of which was found to homologous human fatty acid synthase gene. Relations between these gene mutations and clinicopathological features will be studied in the future by making antibodies of these spots.

[背景]我们利用高分辨率技术-限制性标志基因组扫描（RLGS），在人肝细胞癌中发现了一个新的基因改变。本研究的目的是使用相同的方法鉴定结直肠癌中新的遗传改变。[方法]从25例大肠癌组织和相应的正常黏膜组织中提取高分子量的基因组DNA。用限制酶Not I切割各DNA，通过使用Pvu II的第一维电泳进行大小分级，然后用作为凝胶中的第三种酶的Pst I切割DNA片段，并通过二维电泳分离。通过比较每对RLGS图谱，我们发现了6个共同的变化点。直接从凝胶中克隆最频繁变化的斑点。25例结直肠癌中19例（76.0%）出现相同的斑点状改变，而正常粘膜未见此改变。克隆了与该点对应的410 bp DNA片段。发现该序列与人脂肪酸合成酶基因（FAS）的部分同源性为69%，并且定位于人草图序列的第3染色体的95%。3个克隆点（点B、C和F）分别在44%（11/25）、24%（6/25）和28%（7/25）的病例中观察到，它们分别为271、168和374 bp的DNA片段。来自斑点B、C和F的DNA序列也分别与第一和第九染色体的基因组重叠群以及人Not I克隆同源。[结论]克隆了4个位点，其中1个与人脂肪酸合成酶基因同源。这些基因突变与临床病理特征之间的关系将在未来通过制备这些点的抗体来研究。