Regulartory Mechanism of Ovarian Aromatase by Nuclear Receptor, PPARγ : RXR

核受体 PPARγ 调节卵巢芳香酶的机制：RXR

• 批准号: 13671157
• 负责人: YANASE Toshihilo
• 金额: $ 2.62万
• 依托单位: Kyushu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13671157/
• 关键词: ovary; granulosa cell; aromatase; PPARγ; RXR

Our previous studies demonstrated that PPARgamma specific-ligand troglitazone (TGZ) and/or RXR specific ligand LG100268 (LG) decreased the P450 aromatase activity in cultured human ovarian granulosa cells as well as in the human granulose-like tumor KGN cells. At the present study we evidenced that TGZ+LG decreased P45Oarom promoter II in both the ovarian KGN cells and the fibroblast NIH-3T3 cells in a PPARgamma dependent manner. Further, the TGZ+LG inhibition to both aromatase activity and the P450arom promoter II was completely eliminated when NF-kappaB was blocked, suggesting NF-kappaB, which is endogenously expressed in both fibroblast and the granulose cells, might be the mediator of the inhibition. On the other hand, activation of NE-kappaB either by a direct cotransfection of P65 subunit or by transfection of NIK to activate the endogenous NF-kappaB, was found to up-regulate P450arom promoter II activity. Though activation of PPARgamma.RXR could not interfere with endogenous expression of IkappaBalpha nor that of P65, it did impair the transactivation ability of NF-kappaB, as documented by a NF-kappaB elements containing luciferase reporter, which was dose-dependently inhibited by IGZ+LG.Our data reinforce the potential use of synthetic PPARgamma and RXR ligands for diseases like breast cancer where estrogen plays prominent roles. And besides the classical P450aorm modulators like PKA, SF-1, nuclear receptors like PPARgamma, RXR and their cross-talk with NF-kappa B may also have important roles, wherein, PPARgama inhibits P450arom gene through NF-kappaB.

我们以前的研究表明，PPARgamma特异性配体曲格列酮（TGZ）和/或RXR特异性配体LG 100268（LG）降低培养的人卵巢颗粒细胞以及人颗粒样肿瘤KGN细胞中P450芳香化酶活性。在本研究中，我们证明TGZ+LG以PPARgamma依赖的方式降低卵巢KGN细胞和成纤维细胞NIH-3 T3细胞中的P45 Oarom启动子II。此外，TGZ+LG对芳香化酶活性和P450 arom启动子II的抑制在NF-κ B被阻断时被完全消除，这表明在成纤维细胞和颗粒细胞中内源性表达的NF-κ B可能是抑制的介质。另一方面，通过直接共转染P65亚基或通过转染NIK激活内源性NF-κ B来激活NE-κ B，发现上调P450 arom启动子II活性。虽然激活PPARgamma.RXR不能干扰IkappaB α和P65的内源性表达，但它确实损害了NF-κ B的反式激活能力，如含有荧光素酶报告基因的NF-κ B元件所证明的，IGZ+LG剂量依赖性地抑制了NF-κ B元件的表达。我们的数据加强了合成PPARgamma和RXR配体在乳腺癌等疾病中的潜在用途，其中雌激素起着重要作用。除了经典的P450 arom调节剂PKA、SF-1外，核受体PPARgamma、RXR及其与NF-κ B的相互作用也可能发挥重要作用，其中PPARgamma通过NF-κ B抑制P450 arom基因。

项目成果

Fujii A, Harada T, Yamauchi N, Iwabe T, Nishi Y, Yanase I, Nawata H, Terakawa N.: "Interleukin-8 gene and protein expression are up-regulated by intenleukin-1 beta in normal human ovarian cells and a granulosa tumor cell line."Fertil Steril.. 79. 151-157

Fujii A、Harada T、Yamauchi N、Iwabe T、Nishi Y、Yanase I、Nawata H、Terakawa N.：“在正常人卵巢细胞和颗粒细胞中，白介素 8 基因和蛋白表达被白细胞介素 1 β 上调

Mu Y, Yanase T, Nishi Y et al.: "Saturated free fatty acids, palmitic acid and stearic acid induces apoptosis of human granulosa cells"Endocrinology. 142. 3590-3597 (2001)

Mu Y、Yanase T、Nishi Y 等人：“饱和游离脂肪酸、棕榈酸和硬脂酸诱导人颗粒细胞凋亡”内分泌学。

Mu Y-M, Yanase T, Nishi Y, Takayanagi R, Goto K, Nawata H: "Nuclear receptor system constituted by PPARg:RXR heterodimers inhibits the expression of cytochrome P450arom in human granulosa cells"Mol Cell Endocrinol. 181. 239-248 (2001)

Mu Y-M、Yanase T、Nishi Y、Takayanagi R、Goto K、Nawata H：“由 PPARg:RXR 异二聚体构成的核受体系统抑制人颗粒细胞中细胞色素 P450arom 的表达”Mol Cell Endocrinol。

Fujii A, Harada T, Yamauchi N, Iwabe T, Nishi Y, Yanase T, Nawata H, Terakawa N: "Interleukin-8 gene and protein expression are up-regulated by interleukin-1beta in normal human ovarian cells and a granulosa tumor cell line."Fertil Steril.. 79. 151-157 (2

Fujii A、Harada T、Yamauchi N、Iwabe T、Nishi Y、Yanase T、Nawata H、Terakawa N：“在正常人卵巢细胞和颗粒瘤细胞中，IL-8 基因和蛋白质表达被 IL-1β 上调

Saitoh M, Yanase T, et al.: "Tributyltin or Triphenyltin Inhibits Aromatase Activity in the Human Granulosa-like Tumor Cell Line KGN"Biochem Biophys Res Commun. 289. 198-204 (2001)

Saitoh M、Yanase T 等人：“三丁基锡或三苯基锡抑制人颗粒样肿瘤细胞系 KGN 中的芳香酶活性”Biochem Biophys Res Commun。