ANALYSIS OF CELL GROWTH AND TRANSFORMATION BY ORNITHINE DECARBOXYLASE

鸟氨酸脱羧酶分析细胞生长和转化

• 批准号: 09670566
• 负责人: SHIOMI Susumu
• 金额: $ 1.79万
• 依托单位: THIRD DEPARTMENT OF INTERNAL MEDICINE, OSAKA CITY UNIVERSITY MEDICAL SCHOOL
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670566/
• 关键词: ORNITHINE DECARBOXYLASE; DFMO; HepG2 cell; DIFFERNTIAL DISPLAY; POLYAMINE; PUTRESCINE; cytochrome oxidase; Hep G2細胞; Cytochrome oxidase; Differential Display法; ODC遺伝子; Nuclease Protection Assay; ポリアミン代謝; Differential dysplay法; PEST領域; SSCP法

In order to identify gene differentially expressed by putrescine, a polyamine, which play important roles in the regulation of cell proliferation and the development of cancer, we performed mRNA differential display analysis using total RNA extracted from HepG2 cells (human hepatoblastoma cell line) treated with a specific inhibitor of polyamine biosynthesis, α-difluoromethyl ornithine (DFMO). A total of 25 genes were up-regulated and 32 genes down-regulated by putrescine. Of the genes differentially expressed by putrescine, we chose three that were related to the respiratory chain and oxidative phosphorylase and analyzed them by Northern blot analysis. Cytichrome oxidase submit 1, low molecular mass ubiquinone- binding protein, and NADH dehydrogenase subunit 2 were found to down- regulated by putrescine. We examined intracellular ATP level in Hep G2 cells, and found that ATP level in DFMO- treated cells was increased by exogenous putrescine.

为了鉴定在细胞增殖调节和癌症发展中发挥重要作用的多胺腐胺差异表达的基因，我们使用从经多胺生物合成特异性抑制剂α-二氟甲基鸟氨酸（DFMO）处理的HepG2细胞（人肝母细胞瘤细胞系）中提取的总RNA进行了mRNA差异显示分析。腐胺总共上调了 25 个基因，下调了 32 个基因。在腐胺差异表达的基因中，我们选择了三个与呼吸链和氧化磷酸化酶相关的基因，并通过 Northern blot 分析对它们进行了分析。发现腐胺下调细胞色素氧化酶提交 1、低分子量泛醌结合蛋白和 NADH 脱氢酶亚基 2。我们检查了 Hep G2 细胞中的细胞内 ATP 水平，发现 DFMO 处理的细胞中 ATP 水平因外源腐胺而增加。

项目成果

藤本俊輔他: "Differential displayによる肝細胞増殖関連遺伝子の解析"第3回日本肝臓学会総会.

Shunsuke Fujimoto等人：“通过差异显示分析与肝细胞增殖相关的基因”，日本肝病学会第三届年会。

Tanaka T, Yamashita T, Goto Y, Kondo K, Monna T, Nishiguchi S, Kuroki T, Otani S.: "Relationship between glutathione and polyamine metabolism after partial hepatectomy in alcohol-treated rats."Alcohol Clin Exp Res. 22. 125-128 (1998)

Tanaka T、Yamashita T、Goto Y、Kondo K、Monna T、Nishiguchi S、Kuroki T、Otani S.：“酒精治疗大鼠部分肝切除术后谷胱甘肽和多胺代谢的关系。”酒精临床实验研究。

Kubo S, et al.: "Relationship of polyamine metabolism to degee of malignancy of human hepatocellular carcinoma."Oncology Reports. 5. 1385-1388 (1998)

Kubo S 等人：“多胺代谢与人类肝细胞癌恶性程度的关系。”肿瘤学报告。

Kuroda T, Nishiguchi S, Kuroki T, Otani S.: "Effect of alanine on polyamine metabolism in hepatocytes with cellular injury induced by D-galactosamine"Hepatol Res. 12. 31-44 (1998)

Kuroda T、Nishiguchi S、Kuroki T、Otani S.：“丙氨酸对 D-半乳糖胺诱导的细胞损伤肝细胞中多胺代谢的影响”Hepatol Res。

Tanaka T et al.: "Relationship between glutathion and polyamine metabolism after partial hapatectomy in alcohol-treated rats." Alcoholism:Clinical and Experimental Reaserch. 22. 125-128 (1998)

Tanaka T 等人：“酒精治疗大鼠部分肝切除术后谷胱甘肽和多胺代谢之间的关系。”