The Development of novel therapeutic strategy for anaplastic thyroid carcinoma by manipulating the ubiquitin-proteasome activity

通过操纵泛素蛋白酶体活性开发未变性甲状腺癌的新治疗策略

• 批准号: 12671088
• 负责人: TANIGUCHI Shin-ichi
• 金额: $ 2.18万
• 依托单位: Tottori University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671088/
• 关键词: UBIQUITIN; PROTEASOME; ANAPLASTIC CARCINOMA; THYROID; プロテアソーム活性化因子; Tet onシステム

Anaplastic thyroid carcinoma is a poorly differentiated carcinoma found in elderly people. The patients with anaplastic carcinoma always suffer from poor prognosis. It is an urgent business to develop the effective strategy to suppress the growth of anaplastic carcinoma and prolong the prognosis of patients. We focused on the ubiqitin-proteasome system. Proteasome is a major intracellular proteinase found as a large protein complex composed of at least 14 distinct but homologous subunits with molecular masses of 21-32 kD and they are assembled into an approximately 700 kDa cylindcal structure. It is involved in the destruction of regulatory proteins responsible for biological processes such as cell cycle progression. We analyzed the expression of proteasome subunits in anaplastic thyroid carcinoma and found that proteasome subunit C2 and proteasome activator g (PA28g) are highly expressed in its rapid-growing cancer cells. This result is repeatedly confirmed by western blotting and imm … More unocytochemistry, when other types of thyroid carcinoma were also estimated. Interestingly, PA 28g is localized in nuclei of cancer cells, suggesting PA28g could be one of the good candidate for manipulating the growth of cancer cells. We then analyzed the detailed turn-over of PA28g using rat functional thyroid cell line, FRTL5 cells. The growth-stimuli, TSH and insulin, induced PA28-γ expression in FRTL5 cell. Intere stingly, both treatment not only upregulate PA28-γ expression but also recruite PA28-γ from cytosol to nucleus. The combination of TSH and insulin showed highest inducibility of PA28-γ as well as dual DNA synthesis of FRTL5. Proteasome inhibitors such as lactacystin significantly blocked the DNA synthesis induced by TSH and insulin, indicating the intrinsic 20S proteasome is involved in thyroid cell growth. Thus, our results indicate PA28-γ is induced by goitrogenic factors, TSH and insulin, in thyroid cell and potentially contribute to thyroid cell growth by potentiating 20S proteasome activity.Taken together, overexpressed proteasome component such as C2 or PA28-γ will be good candidates for manipulating proteasome activity, thereby interfering with anaplastic cancer cell growth. Less

间变性甲状腺癌是一种在老年人中发现的低分化癌。间变性癌患者预后较差。寻找有效的策略来抑制未分化癌的生长，延长患者的预后是当务之急。我们重点研究了泛素-蛋白酶体系统。蛋白酶体是一种主要的胞内蛋白水解酶，由至少14个不同但同源的亚基组成，分子质量为21-32kD，它们组装成一个约700 kDa的圆柱形结构。它参与破坏负责细胞周期进程等生物过程的调节蛋白。我们分析了蛋白酶体亚基在间变性甲状腺癌中的表达，发现蛋白酶体亚基C2和蛋白酶体激活物g(PA28g)在其快速生长的癌细胞中高表达。这一结果被Western blotting和IMM…反复证实。更多的单细胞化学，当其他类型的甲状腺癌也被估计时。有趣的是，PA28g定位于癌细胞的细胞核中，提示PA28g可能是调控癌细胞生长的良好候选者之一。然后，我们使用大鼠功能甲状腺细胞系FRTL5细胞分析了PA28g的详细翻转。促甲状腺激素和胰岛素可诱导FRTL5细胞PA28-γ的表达。有趣的是，两种治疗方法不仅上调了PA28-γ的表达，而且使PA28-γ从胞浆向胞核聚集。促甲状腺激素和胰岛素的联合使用对PA28-γ的诱导率最高，对FRTL5DNA的合成具有双重诱导性。蛋白酶体抑制剂lactacystin显著阻断TSH和胰岛素诱导的DNA合成，提示固有的20S蛋白酶体参与了甲状腺细胞的生长。因此，我们的结果表明，PA28-γ是由促甲状腺激素和胰岛素在甲状腺细胞中诱导的，并可能通过增强20S蛋白酶体的活性而促进甲状腺细胞的生长。综上所述，过表达的蛋白酶体成分如C2或PA28-γ将很好地调控蛋白酶体的活性，从而干扰间变性癌细胞的生长。较少

项目成果

Shin-ichi Taniguchi, Tomohisa Okamura, Yuka Santo, Hiroko Fukui, Hideki Shimizu, Akio Yoshida, Yoshihiko Ueta and Chiaki Shigemasa: "Proteasome activator 28-γ (PA28-γ) is induced by goitrogenic factors-TSH and insulin in thyroid cell"Journal of Clinical E

Shin-ichi Taniguchi、Tomohisa Okamura、Yuka Santo、Hiroko Fukui、Hideki Shimizu、Akio Yoshida、Yoshihiko Ueta 和 Chiaki Shigemasa：“蛋白酶体激活剂 28-γ (PA28-γ) 由甲状腺细胞中的致甲状腺肿因子 - TSH 和胰岛素诱导”临床电子杂志

T. Okamura, S-I. Taniguchi, A. Yoshida, H. Shimizu, M. Sakai1, H. Maeta, H. Fukui, Y. Ueta and C. Shigemasa: "Abnormally high expression of Proteasome Activator-γ (PA28-γ) in Thyroid Neoplasma"Journal of Clinical Endocrinology & Metabolism. (in submission

T. Okamura、S-I. Taniguchi、A. Yoshida、H. Shimizu、M. Sakai1、H. Maeta、H. Fukui、Y. Ueta 和 C. Shigemasa：“蛋白酶体激活剂-γ (PA28-γ) 的异常高表达甲状腺肿瘤”临床内分泌与代谢杂志。