CLONING OF NOVEL GENE WHOSE PRODUCT MEDIATES LIGAND-INDEPENDENT INDUCTION OF CYP1A1

其产物介导 CYP1A1 配体独立诱导的新基因的克隆

• 批准号: 12480153
• 负责人: KIKUCHI Hideaki
• 金额: $ 6.66万
• 依托单位: TOHOKU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12480153/
• 关键词: Omeprazole; Ah receptor; somatic cell hybrid; CYP1A1; Reverse FISH; ダイオキシン; リガンド非依存性; ベンズイミダゾール; 細胞融合

Our aim of this project is to identify and clone gene whose product mediates ligand-independent induction of CYP1A1 Previously, we showed that CYP1A1 expression can be induced by omeprazole (OP) in the human cell line HepG2,but not in the mouse cell line Hepa-1 cells. Now we show induction of CYP1A1 by α-naphthoflavone (αNF) in Hepa-1 cells. This induction was inhibited by the tyrosine kinase inhibitor herbimycin A, but not by the aromatic hydrocarbon (Ah)-receptor antagonist, suggesting the presence of a ligand-independent signal-transduction pathway in the mouse cell line too. We utilized the lack of CYP1A1 induction by OP in Hepa-1 cells to map a putative human gene OP-responsiveness in cell hybrids produced by fusion of Hepa-1 and HepG2 cells. OP-induced CYP1A1 expression was detected in four out of the 32 Hepa-1xHepG2 cell hybrids analyzed. To help identity the gene locus, a radiation-hybrid cell (E11 )was constructed Use of reverse-fluorescence in situ hybridization revealed that these five cell lines commonly retained human chromosome 10p. These results suggest that the human gene for OP-responsiveness is present on chromosome 10p13.

本课题的目的是鉴定和克隆CYP 1A 1非配体依赖性诱导表达的基因。以前，我们发现奥美拉唑（omeprazole，OP）可以诱导人肝癌细胞系HepG 2中CYP 1A 1的表达，但不能诱导小鼠肝癌细胞系Hepa-1中CYP 1A 1的表达。现在，我们显示α-萘甲酮（αNF）在Hepa-1细胞中诱导CYP 1A 1。这种诱导抑制酪氨酸激酶抑制剂除莠霉素A，但不是由芳烃（Ah）受体拮抗剂，这表明在小鼠细胞系中存在的配体独立的信号转导通路太。我们利用Hepa-1细胞中OP对CYP 1A 1诱导的缺乏，在Hepa-1和HepG 2细胞融合产生的细胞杂交体中绘制了一个推定的人类基因OP-反应性。在分析的32个Hepa-1xHepG 2细胞杂交体中的4个中检测到OP诱导的CYP 1A 1表达。为进一步鉴定该基因的位置，我们构建了一个辐射杂交细胞（E11）。反向荧光原位杂交结果显示，这5个细胞系均保留了人类染色体10 p。这些结果表明，OP-反应性的人类基因存在于染色体10 p13上。

项目成果

Kikuchi, H. et al.: "Presence of omeprazole-sensitive gene product which is responsible for CYP1A1 induction in human HepG2 cells."Drug Metabolism Reviews. 32. 69-69 (2000)

Kikuchi, H. 等人：“奥美拉唑敏感基因产物的存在负责人 HepG2 细胞中 CYP1A1 的诱导。”药物代谢评论。

Kikuchi, H.: "Method for evaluation of immunotoxicity of dioxin compounds using human T-lymphoblastic cell line, L-MAT."Chemosphere. 43. 815-818 (2001)

Kikuchi, H.：“使用人 T 淋巴细胞系 L-MAT 评估二恶英化合物免疫毒性的方法。”Chemosphere。

Kikuchi,H. et.al.: "Presence of omeprazole-sensitive gene product which is responsible for CYP1A1 induction in human HepG2 cells."Drug Metabolism Reviews,. 32. 69-69 (2000)

菊池，H.

Itaru Ishida, Hiroshi Kubo, et al.: "Hypoxia Diminishes Toll-Like Receptor 4 Expression Through Reactive Oxygen Species Generated by Mitochondria in Endothelial Cells."The Journal of Immunology. 169. 2069-2075 (2002)

Itaru Ishida、Hiroshi Kubo 等人：“缺氧通过内皮细胞线粒体产生的活性氧减少 Toll 样受体 4 的表达。”《免疫学杂志》。

Shibazaki, M., Takeuch, T.et al.: "Suppression by p38 MAP kinase inhibitors (pyridinyl imidazole compounds) of Ah receptor target-gene activation by TCDD, and the possible mechanism."J.Biol.Chem.. 279. 3869-3876 (2004)

Shibazaki, M., Takeuch, T.等人：“p38 MAP 激酶抑制剂（吡啶基咪唑化合物）对 TCDD 对 Ah 受体靶基因激活的抑制，以及可能的机制。”J.Biol.Chem.. 279。