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

分子克隆鉴定出编码α_1-肾上腺素受体的3个不同的cDNA(α_1-肾上腺素受体1a&gt；、α_1b&gt；和α_1-1d&gt；)。在亚型中，人类前列腺主要表达α-肾上腺素能受体，这些肾上腺素能受体介导收缩反应。已知人类存在Pst I限制性内切酶消化的肾上腺素能受体基因的两个限制性片段长度多态性(RFLP)。这种多态可能在α-肾上腺素受体的羧基末端增加了一个棕榈酰化位点，暗示了异常的信号转导机制。此外，肾上腺素能受体第三个胞质环的突变也可能导致受体/信号转导的结构性激活。在目前的研究中，我们寻找了α1a和gt；肾上腺素能受体基因的第三细胞内环中可能的突变。27例患者手术时获得新鲜前列腺组织，冰冻于液氮中，S…在-80ﾟC温度下，从前列腺组织中提取总RNA，逆转录，制备成c DNA。用DNA测序仪对编码肾上腺素能受体胞内第三环的聚合酶链式反应产物片段进行纯化和分析。结果，α-肾上腺素能受体基因的第三个胞质环未发生突变。此外，我们还研究了肾上腺素能受体基因Pst I-RFLP是否与女性尿路综合征有关。从基因组DNA中扩增出502bp的扩增产物，其中含有Arg492的DNA序列。用Pst I限制性内切酶对正常对照组和尿路综合征中年妇女进行了α_1a和α_1a和α_肾上腺素能受体基因的2个RFLP的检测。在日本，尿路综合征患者和正常人的α-肾上腺素能受体基因频率分布差异无统计学意义。然而，这种多态在日本和美国正常人群中的频率分布明显不同。我们的初步研究没有提供任何证据支持肾上腺素能受体基因多态性与蕨类尿路综合征之间的关联，然而，日本和美国都需要进行更大规模的研究。较少

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