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]我们通过使用高分辨率技术，限制性地标基因组扫描（RLGS）确定了人肝细胞癌的新遗传改变。这项研究的目的是使用相同的方法鉴定结直肠癌的新遗传改变。[方法]从25种结直肠癌组织和相应的正常粘膜中提取高分子量基因组DNA。将每个DNA用限制酶裂解，而不是I，使用PVU II通过一维电泳分离的大小，然后将DNA片段通过PST I裂解为凝胶中的第三个酶，并通过2维电泳分离。通过比较每对RLGS轮廓，我们检测到6个常见的更改点。最常见的斑点直接从凝胶中克隆。 25（76.0％）结直肠癌病例中有19例显示出相同改变的斑点，在正常粘膜中从未出现过。克隆了与该位置相对应的410 bp DNA片段。发现该序列与部分人脂肪酸合酶基因（FAS）同源69％，并将其映射到人类草稿序列的第三染色体的95％。在44％（11/25），24％（6/25）和28％（7/25）中观察到三个克隆斑点（B，C和F），分别为271-，168-和374-BP DNA片段。来自斑点B，C和F的DNA序列也与第一和第九染色体的基因组重叠群同源，分别是人类不是I克隆。[结论]四个斑点被克隆了，其中一个被发现是同源的人脂肪酸合酶基因。这些基因突变与临床病理学特征之间的关系将在未来通过制作这些斑点的抗体来研究。