The analysis of molecular mechanism for IRS-1 ubiquitination by nitric oxide

一氧化氮泛素化IRS-1的分子机制分析

• 批准号: 18591517
• 负责人: SUGITA Hiroki
• 金额: $ 2.53万
• 依托单位: Kumamoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18591517/
• 关键词: nitric oxide; TRS(insulin receptor substrate)-1; IGF(insulin like growth factor)-1; insulin; ubiquitin; Akt; pancreatic cancer; GSK(glvcogen svnthase kinase)-3; insulin receptor substrate-1; ユビキチン化; 膵癌細胞; insulin like growth factor-1; 一酸化窒素(NO

(1) The effects of nitric oxide on IGF-R/1RS-1 /PI3-K/Akt pathway and MAPK pathway in cancer cells.Nitric oxide (NO) donor down-regulated insulin/IGF signal including IRS-1, Akt, and GSK-3 in MIAPaCa-2 cells (derived from pancreatic cancer). On the other hand, NO donor up-regulated the phosphorylation of Erk 1/2.(2) IRS-1 ubiquitination by NO donorProtease inhibitor completely inhibited the degradation of IRS-1 protein by NO in MIAPaCa-2. The result suspected that NO regulated the ubiquitination and the degradation of IRS-1 protein in MIAPaCa-2. We produced the some constructs of IRS-1 delation mutants, and made the MIAPaCa-2 express them. Then, we found out the important site for IRS-I protein degradation by NO.(3) The effects of NO on cancer proliferationNO down-regulated the cell growth in some cancer cell lines.We performed the cloning of stable cell lines; MIAPaCa-2 over-expressing IRS-1 wild type protein, MIAPaCa-2 over-expressing IRS-1 dominant negative. The cell line, over-expressing IRS-1 wild type protein, show high seed for cell growth, and greater sensitivity for NO. On the other hand, cell line, over-expressing IRS-1 dominant negative, show low speed of cell growth, and smaller sensitivity for NO. These results indicates that NO down-regulates IGF signal by the depression of IRS-1 protein. That may be one of the mechanisms for the down-regulation of cancer proliferation by NO.

(1)一氧化氮（NO）对胰腺癌MIAPaCa-2细胞胰岛素样生长因子（IGF）信号通路（IRS-1、Akt和GSK-3）的影响。另一方面，NO供体上调Erk 1/2的磷酸化。(2)IRS-1被NO供体蛋白酶抑制剂泛素化完全抑制了MIAPaCa-2中IRS-1蛋白被NO降解。本研究结果推测NO对MIAPaCa-2细胞IRS-1蛋白的泛素化和降解有调节作用。我们构建了IRS-1缺失突变体，并在MIAPaCa-2中进行了表达。从而找到了NO降解IRS-I蛋白的重要位点。(3)NO对肿瘤细胞增殖的影响NO对某些肿瘤细胞系的生长有负调控作用，我们克隆了稳定的细胞系：MIAPaCa-2过表达IRS-1野生型蛋白，MIAPaCa-2过表达IRS-1显性阴性。过表达IRS-1野生型蛋白的细胞系显示出细胞生长的高种子，并且对NO具有更高的敏感性。另一方面，过表达IRS-1显性阴性的细胞系显示出细胞生长的低速度，并且对NO具有更低的敏感性。这些结果表明，NO通过抑制IRS-1蛋白来下调IGF信号。这可能是NO下调肿瘤增殖的机制之一。

项目成果

Combined chemotherapy of irinotecan and low-dose cisplatin (I/low-P) against metastatic biliary tract cancer; report of consecutive three cases

伊立替康与小剂量顺铂（I/low-P）联合化疗治疗转移性胆道癌；

• 发表时间: 2006
• 期刊: J Hep Bil Pancr Surg 13
• 作者: Maehara;N.;Chijiiwa;K.;Eto;T.;Funagayama;M.;Uchiyama;S.;Nakashima;S.;Hidaka;H.;Hotokezaka;M.;Ohmuraya M;Takamori H;Motomura Y;Komori H;Masuda Y;Chikamoto A;Okuma T;Maeda K;Maeda K;Hosaka S;Hirota M;Sugita H
• 通讯作者: Sugita H

Combined radical retropubic prostatectomy and abdominoperineal excision of the rectum for locally invasive rectal cancer as a less invasive surgery: report of a case.

联合根治性耻骨后前列腺切除术和腹会阴直肠切除术作为一种微创手术治疗局部浸润性直肠癌：病例报告。

• 发表时间: 2008
• 期刊: Int. J. Surg. 92
• 作者: Imamura H;Seyama Y;Makuuchi M;Sugita H
• 通讯作者: Sugita H

radical retropubic prostatectomy and abdominoperineal excision of the rectum for locally invasive rectal cancer as a less invasive surgery : report of a case.

根治性耻骨后前列腺切除术和腹会阴直肠切除术作为一种微创手术治疗局部浸润性直肠癌：病例报告。

• 发表时间: 2008
• 期刊: Int. J. Surg. 92(5)
• 作者: Sugita;H.;Egami;H.;Yokoyama;Y.;Suyama;K;Ogawa
• 通讯作者: Ogawa

Chon JY, Tompkins RC, and Martyn. JAA dipocyte apoptosis after burn injury is associated with altered fat metabolism

Chon JY、Tompkins RC 和 Martyn。

• 发表时间: 2006
• 期刊: J Burn Care Res 27(3)
• 作者: Yasuhara;S;Kaneki;M;Sugita;H;Sugita;M;Asai;A;Sahani;N
• 通讯作者: N

dipocyte apoptosis after burn injury is associated with altered fat metabolism

烧伤后双细胞凋亡与脂肪代谢改变有关

• 发表时间: 2006
• 期刊: J Burn Care Res 27
• 作者: Morine;Y;Yasuhara S
• 通讯作者: Yasuhara S