Human alpha_<1a>-adrenoceptor polymorphism : Association with benign prostate hyperplasia

人类α_<1a>-肾上腺素受体多态性：与良性前列腺增生的关联

• 批准号: 09671613
• 负责人: HATANO Akihiko
• 金额: $ 2.3万
• 依托单位: Niigata University
• 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 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671613/
• 关键词: Human; Benign prostate hyperplasis; alpha_<1a>-Adrenoceptor; Polymorphism; α_<LA>-アドレナリン受容体

Molecular cloning identified three distinct cDNAs encoding alpha_1-adrenoceptors (alpha_<1a>, alpha_<1b>, and alpha_<1d>). Among subtypes, human prostate has been shown to express predominantly alpha_<1a>-adrenoceptors, and these mediate the contractile response. It is already known that two restriction fragment length poylmorphisms (RFLPs) of alpha_<1a>-adrenoceptor gene digested with Pst I restriction enzyme exist in human. This polymorphism might confer an additional putative palmitoylation site in the carboxy-terminal segment of alpha_<1a>-adrenoceptor, implying abnormal signal transduction mechanism. Also, mutation in the third cytoplas-mic loop of alpha_<1a>-adrenoceptor might induce the constitutive activation of the receptor/signal transduc-tion. In the present study, we looked for a possible mutation in the third intracellular loop of the alpha_<1a>-adrenoceptor gene. Fresh prostate tissues were obtained at the time of surgery from 27 patients, frozen in liquid nitrogen, and s … More tored at -80ﾟC.Total RNA was extracted from the prostate tissue, then reverse transcribed, and cDNA was prepared. Fragments of PCR products which encodes the third intracellular loop of alpha_<1a>-adrenoceptor were purified and analysed by DNA sequencer. As a result, no mutation in the third cytoplasmic loop of the alpha_<1a>-adrenoceptor gene was observed. Additionary, we examined whether the Pst I-RFLP of the alpha_<1a>-adrenoceptor is associated with female urethral syndrome. PCR product of 502 bp, which contains the DNA sequence corresponding to Arg492, has been amplified from genomic DNA, and 2 RFLPs of alpha_<1a>-adrenoceptor gene, which are digested with Pst I restriction enzyme into 477bp and 25 bp, were cheked in middle aged women of either control or urethral syndrome. The frequency of alpha_<1a>-adrenoceptor polymorphism was not differently distributed between patients with urethral syndrome and normal subjects in Japan. However, the frequency distribution of this polymorphism was significantly different between normal Japanese and U.S.populations. Our preliminary study does not provide any evidence to support an association between alpha_<1a>-adrenoceptor gene polymorphism and fern ale ure-thral syndrome, however, much larger studies of both Japan and U.S.are needed. Less

分子克隆鉴定出三种不同的编码α_1-肾上腺素受体的cDNA（α_<1a>、α_<1b>和α_<1d>）。在前列腺各亚型中，前列腺主要表达α<1a>-肾上腺素能受体，这些受体介导前列腺的收缩反应。已知人α<1a>-肾上腺素能受体基因在Pst Ⅰ酶切后存在两种限制性片段长度多态性。这种多态性可能使α-肾上腺素受体羧基端增加了一个棕榈酰化位点<1a>，提示信号转导机制异常。α-肾上腺素能受体第三环的突变也<1a>可能引起受体的组成性激活/信号转导。在本研究中，我们在α-肾上腺素受体基因的第三胞内环中寻找可能的突变<1a>。手术时从27名患者中获取新鲜前列腺组织，冷冻在液氮中， ...更多信息 从前列腺组织中提取总RNA，然后反转录，制备cDNA。PCR产物<1a>经纯化和DNA测序分析，得到编码α-肾上腺素能受体第三环的片段。结果表明，在α-肾上腺素能受体基因的第三胞质环中没有<1a>观察到突变。此外，我们还研究了α-肾上腺素能受体Pst Ⅰ-RFLP是否<1a>与女性尿道综合征有关。从尿道综合征患者基因组DNA中扩增出502 bp的PCR产物，其中含有与Arg 492相对应的DNA序列。<1a>用Pst Ⅰ限制性内切酶将α-肾上腺素能受体基因的两个片段分别切为477 bp和25 bp，检测两个片段的限制性内切酶多态性。α-<1a>肾上腺素受体多态性的频率在日本尿道综合征患者和正常人之间的分布没有差异。然而，这种多态性的频率分布是显着不同的正常日本和美国人口。我们的初步研究没有提供任何证据支持α-<1a>肾上腺素能受体基因多态性与蕨类植物尿道综合征之间的关联，然而，需要在日本和美国进行更大规模的研究。少

项目成果

Takako Okamoto-Koizumi: "Pharmacological and molecular biological evidence for ET_A endothelin receptor subtype me-diating mechanical responses in the detrysor smooth muscle of the human urinary bladder" Clinical Science. 96(印刷中). (1999)

Takako Okamoto-Koizumi：“ET_A 内皮素受体亚型介导人类膀胱平滑肌机械反应的药理学和分子生物学证据”《临床科学》96（出版中）。

Masayuki Takeda: "Evidence for β_3-adrenoceptor subtypes in relaxation of the human urinary blad-der detrusor : analysis by molecular biological and phrmacological methods" Journal of pharmacology and experimental therapeutics. 288(3)印刷中. (1999)

Masayuki Takeda：“β_3-肾上腺素受体亚型在人膀胱逼尿肌松弛中的证据：通过分子生物学和药理学方法进行分析”《药理学和实验治疗学杂志》288(3)，出版。

Masayuki Takeda: "Bladder neck suspension using percutaneous bladder neck stabilization to to the pubic bone with a bone-anchor suture fixation system : a new extraperitoneal laparoscopic approach" Urologia Internationalis. 848(印刷中). (1999)

Masayuki Takeda：“使用骨锚缝合固定系统经皮膀胱颈稳定到耻骨的膀胱颈悬吊：一种新的腹膜外腹腔镜方法”Urologia Internationalis 848（出版中）。

Masayuki Takeda, Akihiko Hatano, Kei Arai, Kenji Obara, Toshiki Tsutsui and Kota Takahashi: "13th IUPHAR official satellite symposium : alpha_1-adrenoceptors as targets for therapeutic agents in urology : alpha_1-and alpha_2-adrenoceptors in BPH" European

Masayuki Takeda、Akihiko Hatano、Kei Arai、Kenji Obara、Toshiki Tsutsui 和 Kota Takahashi：“第 13 届 IUPHAR 官方卫星研讨会：α_1-肾上腺素受体作为泌尿科治疗药物的靶点：BPH 中的 α_1-和 α_2-肾上腺素受体”欧洲

Masayuki Takeda, Akihiko Hatano, Shigenori Kurumada, Hitoshi, Syunji Wakatsuki, Hideto Go, Ryusuke Watanabe, Yoshihiko Tomita, Kota Takahashi: "Bladder neck suspension using prcutaneous bladder neck stabilization to to the pubic bone with a bone-anchor su

Masayuki Takeda、Akihiko Hatano、Shigenori Kurumada、Hitoshi、Syunji Wakatsuki、Hideto Go、Ryusuke Watanabe、Yoshihiko Tomita、Kota Takahashi：“膀胱颈悬吊采用经皮膀胱颈稳定，通过骨锚固固定到耻骨上