Analysis of sex hormone-responsive element in sexually dimorphicly expressing CYP2B subfamily gene

CYP2B亚家族基因性二态性表达的性激素反应元件分析

• 批准号: 10672105
• 负责人: NEMOTO Nobuo
• 金额: $ 2.24万
• 依托单位: TOYAMA MEDICAL AND PHARMACEUTICAL UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10672105/
• 关键词: CYP2B; Sex hormone; Glucocorticoid hormone; Phenobarbital; 肝細胞初代培養系; マウス肝細胞初代培養系

CYP2B subfamily constitutively and sexually dimorphicly expresses in the liver. In the mouse, the expression in female is far higher than that in male and the expressed species are different between male (Crp2b10>Cyp2b9) and female (Cyp2b9>Cyp2b10). The expression of these species is inducible by phenobarbital (PB), and the induction mechanism has been extensively revealed at molecular level. However, the mechanism of their constitutive expression has been scarcely investigated.Female and glucocorticoid hormones may participate the expression of Cyp2b9 and Cyp2b10, respecitively, and the latter hormone simultaneously suppresses Cyp2b9 expression. The role of male hormone is also considered to suppress Cyp2b9 expression. The sequence from -2331 to -2281 bp of the 5'-flanking region of Cyp2b10 gene is revealed to be responsible for the estradiol induction using luciferase assay. This region corresponds to the core element of PB-responsive enhancer module (PBREM). Several nucleotide mutations in the PBREM core element show that the NR1 (nuclear receptor 1) site is required for estradiol induction, the same element required for PB induction. The element also plays a critical role for induction by DDT, indicating that NR1 site responds to structurally-diversed inducers. Although Cyp2b9 is a major species expressed in female liver, no information is available for its expression mechanism, but estradiol is found as a potent inducer for investigating of the regulatory pathway.

CYP2B 亚家族在肝脏中组成型和性二态性表达。在小鼠中，雌性中的表达量远高于雄性，且雄性(Crp2b10>Cyp2b9)和雌性(Cyp2b9>Cyp2b10)表达的种类不同。这些物种的表达可由苯巴比妥（PB）诱导，并且诱导机制已在分子水平上得到广泛揭示。然而，其组成型表达的机制却很少被研究。女性激素和糖皮质激素可能分别参与Cyp2b9和Cyp2b10的表达，并且后者同时抑制Cyp2b9的表达。雄性激素的作用也被认为是抑制Cyp2b9的表达。使用荧光素酶测定发现，Cyp2b10 基因 5' 侧翼区域的 -2331 至 -2281 bp 序列负责雌二醇诱导。该区域对应于 PB 响应增强子模块 (PBREM) 的核心元件。 PBREM 核心元件中的几个核苷酸突变表明 NR1（核受体 1）位点是雌二醇诱导所必需的，这与 PB 诱导所需的元件相同。该元素对于 DDT 的诱导也起着关键作用，表明 NR1 位点对结构多样化的诱导剂有反应。虽然Cyp2b9是在女性肝脏中表达的主要物种，但没有关于其表达机制的信息，但发现雌二醇是研究调节途径的有效诱导剂。

项目成果

Jarkumjorn K et al.: "Discriminating activation of CYP2B9 expression in male C57BL/6 mouse liver by β-estradiol."Biochem Biophys Res Commun. 279巻. 288-292 (2000)

Jarkumjorn K 等人：“通过 β-雌二醇区分雄性 C57BL/6 小鼠肝脏中 CYP2B9 表达的激活。”Biochem Biophys Res Commun. 279. 288-292 (2000)

Miyahara T, Harada M, Kozakai A, Matsumoto M, Hashimoto K, Inoue H, Yoda K, Nakatsu T, Kajita S, Yamazaki R, Higuchi S, Kozuka H, Nemoto N.: "Comparison of 26,27-hexafluoro-1α,25-dihydroxyvitamin D_3 and 1α,25-dihydroxyvitamin D_3 on the resorption of bon

宫原T、原田M、小坂井A、松本M、桥本K、井上H、尤达K、中津T、梶田S、山崎R、樋口S、小冢H、根本N.：“26,27-六氟的比较-1α,25-二羟基维生素D_3和1α,25-二羟基维生素D_3对bon吸收的影响

Sakuma T et al.: "Induction of CYP1A2 by phenobarbital in the livers of aryl hydrocarbon-responsive and -nonresponsive mice."Drug Metabol Dispos. 27巻. 379-384 (1999)

Sakuma T 等人：“苯巴比妥在芳烃反应性和非反应性小鼠肝脏中的诱导”，《药物代谢处​​置》，第 27 卷，379-384。

Miyamoto M et al.: "CYP2A6 gene deletion reduces susceptibility to lung cancer."Biochem Biophys Res Commun. 261巻. 658-660 (1999)

Miyamoto M 等人：“CYP2A6 基因缺失降低了对肺癌的易感性。”Biochem Biophys Res Commun. 261. 658-660 (1999)

Chida M, Yokoi T, Nemoto N, Inaba M, Kinoshita M, Kamataki T.: "A new variant CYP2D6 allele(CYP2D6^*21) with a single base insertion in exon 5 in Japanese population associated with a poor metabolizer phenotype."Pharmacogenetics. 9. 287-293 (1999)

Chida M、Yokoi T、Nemoto N、Inaba M、Kinoshita M、Kamataki T.：“一种新的变异 CYP2D6 等位基因 (CYP2D6^*21)，在日本人群的外显子 5 中插入一个单碱基，与代谢不良表型相关。”