SERIAL ANALYSIS OF GENE EXPRESSION (SAGE) IN HUMAN STOMACH CANCER

人类胃癌基因表达 (SAGE) 的系列分析

• 批准号: 11671218
• 负责人: MINAMOTO Toshinari
• 金额: $ 2.18万
• 依托单位: KANAZAWA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671218/
• 关键词: STOMACH CANCER; GENE EXPRESSION; SERIAL ANALYSIS OF GENE EXPRESSION (SAGE)

The purpose of this project is to conduct serial analysis of gene expression (SAGE) for human stomach cancer. Applying SAGE, that was originally developed in the tissue culture system, for global analysis of gene expression in clinical stomach cancer requires purification of high quality mRNA from tissue samples that contain minimal amount of stromal cells. In 1999, prior to examine tissue samples, we analyzed by SAGE for gene expression in a cancer cell line, KKLS that was established from undifferentiated stomach carcinoma in our institute. This preliminary analysis could quantitatively identify 2969 unique genes expressed among 5182 SAGE tags obtained from the cell line, with a linker ratio of 1.02%. Among highly expressed genes were included mitochondrial genes and several unknown genes unregistered to the Gene Bank. We assume that the gene expression profile of KKLS would be fundamental for further analysis of gene expression in human stomach cancer showing different histological types.In 2000, human stomach cancer tissues has been subjected to SAGE.During this analysis we have had difficulty in preparation of highly purified mRNA from the surgical materials, which was later overcome by modification of our method for extraction and purification of mRNA.Similar to the gene expression pattern in KKLS cell line, unknown nuclear genes and mitochondrial genes were quantitatively detected in the library of SAGE tags yielded from human stomach cancer samples. Recent reports showing frequent genetic alteration in mitochondrial DNA suggest an involvement of altered expression of mitochondrial genes in development and progression of stomach cancer. Although we could not complete this project in the given term (1999 to 2000), SAGE for human stomach cancer is being in progress in our laboratory, targeting nuclear as well as mitochondrial genes.

本项目的目的是对人类胃癌进行基因表达序列分析(SAGE)。应用最初在组织培养系统中开发的SAGE，用于临床胃癌基因表达的全球分析，需要从含有最少间质细胞的组织样本中提纯高质量的mRNA。1999年，在检查组织样本之前，我们用SAGE分析了我们研究所从未分化的胃癌建立的癌细胞系KKLS的基因表达。这一初步分析可以在5182个SAGE标签中定量鉴定2969个表达的独特基因，连接子比率为1.02%。在高表达的基因中，包括线粒体基因和几个未在基因库注册的未知基因。我们认为KKLS的基因表达谱将为进一步分析不同组织类型的人胃癌组织的基因表达奠定基础。2000年，我们对人胃癌组织进行了SAGE分析，在此过程中，我们难以从手术材料中制备高纯度的mRNA，后来我们改进了提取和纯化mRNA的方法，克服了这一问题。与KKLS细胞系的基因表达谱相似，从人胃癌样本产生的SAGE标签库中定量检测到了未知的核基因和线粒体基因。最近的报道显示线粒体DNA的频繁遗传改变提示线粒体基因的表达改变参与了胃癌的发生和发展。虽然我们不能在给定的期限内(1999年至2000年)完成这项计划，但我们实验室正在进行针对人类胃癌的SAGE，目标是核基因和线粒体基因。

项目成果

Minamoto T,Mai M,Ronai Z.: "Environmental factors as regulators and effectors of multistep carcinogenesis."Carcinogenesis. 20. 519-527 (1999)

Minamoto T，Mai M，Ronai Z.：“环境因素作为多步致癌作用的调节器和效应器。”致癌作用。

Minamoto T, Mai M, Ronai Z.: "K-ras mutation : early detection in molecular diagnosis and risk assessment of colorectal, pancreas and lung cancers-a review"Cancer Detec Prev. 24(1). 1-12 (2000)

Minamoto T、Mai M、Ronai Z.：“K-ras 突变：结直肠癌、胰腺癌和肺癌分子诊断和风险评估的早期检测 - 综述”Cancer Detec Prev。

Minamoto T, Mai M, Ronai Z.: "Environmental factors as regulators and effectors of multistep carcinogenesis"Carcinogenesis. 20(4). 519-527 (1999)

Minamoto T、Mai M、Ronai Z.：“环境因素作为多步致癌作用的调节器和效应器”致癌作用。

Ougolkov A, Mai M, Takahashi Y, Bilim V, Shimizu A, Minamoto T.: "Altered expression of β-catenin and c-erbB-2 in early gastric cancer"J Exp Clin Cancer Res. 19(3). 349-355 (2000)

Ougolkov A、Mai M、Takahashi Y、Bilim V、Shimizu A、Minamoto T.：“早期胃癌中 β-catenin 和 c-erbB-2 的表达改变”J Exp Clin Cancer Res 19(3)。 355 (2000)

Minamoto T,Mai M,Ronai Z: "Environmental factors as regulators and effectors of multistep carcinogenesis"Carcinogenesis. 20・4. 519-527 (1999)

Minamoto T、Mai M、Ronai Z：“环境因素作为多步致癌作用的调节因素和效应因素”致癌作用20・4（1999）。