Suppression subtraction法による消化器疾患の分子病態解析

抑制减法法对胃肠道疾病的分子病理学分析

• 批准号: 13204027
• 负责人: 榎本 信幸
• 金额: $ 4.16万
• 依托单位: Tokyo Medical and Dental University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research on Priority Areas (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 无数据
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13204027/
• 关键词: 消化器疾患; 遺伝子発現; subtraction

(1)生検材料から得られる微量のtotal RNAよりmRNAを特異的にlong distance PCRにより増幅するSMART PCR法を導入し、lugのtotal RNAから数十microgramのmRNA由来のcDNAを得ることが可能となった。さらに、suppressive subtraction法による高効率のsubtraction cloningを行い、疾患特異的発現をする遺伝子のPCR産物を得ることができた。(2)肝癌cDNAと周囲非癌部cDNAをsubtractionすることにより、FAK, GS, DCC, Gia, PGC, decorinなどの遺伝子の肝癌における特異的な発現変化を見いだした。(3)大腸癌から周辺正常大腸粘膜のsubtractionでは、同様にCEA, NCA, Bub1, Maspin, T0G, GABA-R, dkk4などの遺伝子の発現を確認した。(4)慢性C型肝炎および自己免疫性肝炎ではchemokineであるIP-10が特異的に発現していた。(5)原発性胆汁性肝硬変では種々のミトコンドリア由来遺伝子が発現していることを見いだした。最近、消化器疾患においても遺伝子発現変化の解析にmicroarrayが導入されているが、個々の解析結果は多様であり、これらの疾患の病態の全体像を解明しうる普遍的な結果は未だ得られていないと思われる。本研究で同定された遺伝子変化もこれらのmicroarrayでの結果とは異なるものもあり、当面、これらの解析手法は相互に補完的な情報をもたらすと考えられる。一方、本研究で慢性肝炎で発現が上昇していることが明らかとIP-10は急性C型肝炎動物モデルのmicroarray解析でも最も発現変化の大きな遺伝子の一つであり、病態の解明に重要な分子と考えらる。

(1) the raw material is used to obtain trace amounts of total RNA, mRNA, long distance PCR, SMART PCR, lug, total RNA, and dozens of micrograms of mRNA are derived from cDNA. The results of suppressive subtraction method and suppressive subtraction method showed that the rate of subtraction cloning was high, and that of patients with special diseases was higher than that of patients with diseases. (2) cDNA, subtraction, DCC, Gia, PGC, decorin in the non-cancerous part of the cDNA week. (3) subtraction positive and homologous CEA, NCA, Bub1, Maspin, T0G, GABA-R, dkk4 were found in the normal mucosa of patients with cancer. (4) chronic hepatitis C, autoimmune hepatitis, chemokine hepatitis, IP-10, chronic hepatitis C, autoimmune hepatitis, chronic hepatitis C, chronic hepatitis C, autoimmune hepatitis, chronic hepatitis C, chronic hepatitis C, autoimmune hepatitis, chronic hepatitis C, chronic hepatitis C, autoimmune hepatitis, chronic hepatitis C, chronic hepatitis C, autoimmune hepatitis, chronic hepatitis C. (5) the etiology of various types of biliary cirrhotic liver disease is different from that of the original biliary cirrhosis. Recently, the digestive tract disease patients were diagnosed and analyzed. The results of microarray analysis showed that the symptoms of multiple diseases and diseases of digestive diseases were common. The results showed that the results did not meet the needs of the general population. The purpose of this study is to determine the results of this study. The results show that the results of this study are similar to the results of this study. In the same way, the results of this study are similar to the results of this study. In this study, the results of this study are as follows: in this study, the results of this study are similar to the results of this study. On the one hand, in the current study of chronic hepatitis, we need to understand that IP-10 is the most important molecule in the study of acute hepatitis C. microarray analysis is the most effective way to realize the disease.

项目成果

Nagayama K: "Overexpression of interferon gamma-inducible protein 10 in the liver of patients with type I autoimmune hepatitis identified by suppression subtractive hybridization"Am J Gastroenterol. 96. 2211-2217 (2001)

Nagayama K：“通过抑制消减杂交鉴定出 I 型自身免疫性肝炎患者肝脏中干扰素 γ 诱导蛋白 10 的过度表达”Am J Gastroenterol。

Miyasaka Y: "Analysis of differentially expressed genes in human hepatocellular carcinoma using suppression subtractive hybridization"Br J Cancer. 85. 228-234 (2001)

Miyasaka Y：“使用抑制消减杂交分析人肝细胞癌中差异表达的基因”Br J Cancer。