Analysis of TGF-β/BMP signaling during bone metastasis and development of cancer therapy

骨转移过程中TGF-β/BMP信号传导分析及癌症治疗发展

• 批准号: 17390422
• 负责人: IMAMURA Takeshi
• 金额: $ 10.72万
• 依托单位: Japanese Foundation For Cancer Research
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17390422/
• 关键词: cancer; signal transduction; enzyme; 生体分子; 細胞・組織; 転移; 骨; TGF-β; BMP

Members of the transforming growth factor-b (TGF-β) superfamily, including TGF-β and bone morphogenetic proteins (BMPs), are multifunctional cytokines that regulate a wide range of cellular responses, including cell proliferation, differentiation, adhesion, migration, and apoptosis. TGF-β signaling facilitates tumor growth and metastasis in advanced cancer. The TGF-β signaling pathway has correspondingly become an attractive target for drug development in the field of oncology.We investigated the roles of TGF-β and BMP signaling during bone metastasis. We first established a highly bone-metastatic variant of human breast cancer MDA-MB-231 cells, termed MDA-MB-231-5a-D (MDA-231-D). Next, we examined the effects of a novel TGF-β type I receptor (TBR-I) kinase inhibitor, Ki26894, on bone metastasis of MDA-231-D cells. Ki26894 blocked TGF-β signaling in MDA-231-D cells, as detected by suppression of phosphorylation of Smad2 and inhibition of TGF-β-responsive reporter activity. Moreover, Ki … More 26894 decreased the motility and the invasion of MDA-231-D cells induced by TGF-β in vitro. Ki26894 also suppressed transcription of plasminogen activator inhibitor-1 (PAI-1), parathyroid hormone-related protein (PTHrP), and interleukin-11 (IL-11) mRNA of MDA-231-D cells, which were stimulated by TGF-β. X-ray radiography revealed that systemic Ki26894 treatment initiated 1 day before the inoculation of MDA-231-D cells into the left ventricle of BALB/c nu/nu female mice resulted in decreased bone metastasis of breast cancer cells. Moreover, Ki26894 prolonged the survival of mice inoculated with MDA-231-D cells compared to vehicle-treated mice. These findings suggest that TβR-I kinase inhibitors such as Ki26894 may be useful for blocking the progression of advanced cancers.In addition, we identified differentially expressed in chondrocytes 1 (DEC1, also known as SHARP2 and Stral3) as a downstream target of TGF-β signaling, which promotes the survival of breast cancer cells. In the mouse mammary carcinoma cell lines JygMC (A) and 4T1, the TBR-β kinase inhibitors A-44-03 and SB431542 induced apoptosis of cells under serum-free conditions. Oligonucleotide microarray and real-time reverse transcription-PCR analyses revealed that TGF-β induced DEC1 in these cells, and the increase of DEC1 was suppressed by the TβR-I kinase inhibitors as well as by expression of dominant-negative TGF-β type II receptor. Overexpression of DEC1 prevented the apoptosis of JygMC (A) cells induced by A-44-03, and knockdown of endogenous DEC1 abrogated TGF-β-promoted cell survival. Moreover, a dominant-negative mutant of DEC1 prevented lung and liver metastasis of JygMC (A) cells in vivo. Our observations thus provide new insights into the molecular mechanisms governing TGF-β-mediated cell survival and metastasis of cancer. Less

转化生长因子-b (TGF-β) 超家族的成员，包括 TGF-β 和骨形态发生蛋白 (BMP)，是调节多种细胞反应的多功能细胞因子，包括细胞增殖、分化、粘附、迁移和生长。 TGF-β信号通路促进了晚期癌症的肿瘤生长和转移，TGF-β信号通路已成为肿瘤学领域药物开发的一个有吸引力的靶标。我们研究了TGF-β和细胞凋亡的作用。我们首先建立了人乳腺癌 MDA-MB-231 细胞的高骨转移变体，称为 MDA-MB-231-5a-D (MDA-231-D)。接下来，我们检查了其效果。新型 TGF-β I 型受体 (TBR-I) 激酶抑制剂 Ki26894 对 MDA-231-D 细胞骨转移的阻断作用。 MDA-231-D 细胞中的 TGF-β 信号传导，通过抑制 Smad2 磷酸化和抑制 TGF-β 响应报告活性来检测。此外，Ki … 更多 26894 降低了 MDA-231-D 细胞的运动和侵袭。 Ki26894 在体外由 TGF-β 诱导，还可抑制甲状旁腺激素相关蛋白纤溶酶原激活剂抑制剂-1 (PAI-1) 的转录。通过 TGF-β 刺激的 MDA-231-D 细胞的 (PTHrP) 和白介素-11 (IL-11) mRNA，X 射线放射照相显示在接种 MDA-231- 前 1 天开始全身 Ki26894 治疗。将 D 细胞注入 BALB/c nu/nu 雌性小鼠的左心室可减少乳腺癌细胞的骨转移，此外，Ki26894 还可延长小鼠的存活时间。与接种媒介物处理的小鼠相比，接种 MDA-231-D 细胞的小鼠发现 TβR-I 激酶抑制剂（例如 Ki26894）可能有助于阻止晚期癌症的进展。此外，我们还发现了软骨细胞 1 (DEC1) 中的差异表达。 ，也称为 SHARP2 和 Stral3）作为 TGF-β 信号传导的下游靶点，可促进小鼠乳腺癌细胞系中乳腺癌细胞的存活。 JygMC (A) 和 4T1、TBR-β 激酶抑制剂 A-44-03 和 SB431542 在无血清条件下诱导细胞凋亡。寡核苷酸微阵列和实时逆转录 PCR 分析表明，TGF-β 在这些细胞中诱导 DEC1。细胞中，DEC1 的增加受到 TβR-I 激酶抑制剂以及显性失活 TGF-β II 型受体过表达的抑制。 DEC1 阻止 A-44-03 诱导的 JygMC (A) 细胞凋亡，内源性 DEC1 的敲除消除了 TGF-β 促进的细胞存活。此外，DEC1 的显性失活突变体阻止了 JygMC (A) 的肺和肝转移。因此，我们的观察为控制 TGF-β 介导的癌症细胞存活和转移的分子机制提供了新的见解。

项目成果

Visualizing spatiotemporal dynamics of multicellular cell-cycle progression

• DOI: 10.1016/j.cell.2007.12.033
• 发表时间: 2008-02-08
• 期刊: CELL
• 影响因子: 64.5
• 作者: Sakaue-Sawano, Asako;Kurokawa, Hiroshi;Miyawaki, Atsushi
• 通讯作者: Miyawaki, Atsushi

がん研究におけるインビボ光イメージング

癌症研究中的体内光学成像

• 发表时间: 2007
• 作者: Moren A;Imamura T (他3名;2番目);Takeshi Imamura;Takeshi Imamura;今村健志;今村健志;今村健志;今村健志;今村健志;今村健志
• 通讯作者: 今村健志

Arkadia induces degradation of SnoN and c-Ski to enhance transforming growth factor-β signaling

• DOI: 10.1074/jbc.m701294200
• 发表时间: 2007-07-13
• 期刊: JOURNAL OF BIOLOGICAL CHEMISTRY
• 影响因子: 4.8
• 作者: Nagano, Yoshiko;Mavrakis, Konstantinos J.;Miyazawa, Keiji
• 通讯作者: Miyazawa, Keiji

The ALK-5 inhibitor A-83-01 inhibits Smad signaling and epothelial-to-mesenchymal transition by transforming growth factor-β.

ALK-5 抑制剂 A-83-01 通过转化生长因子-β 抑制 Smad 信号传导和上皮间质转化。

• 发表时间: 2005
• 期刊: Cancer Sci. 96
• 作者: Tojo M;Imamura T (他6名;8番目)
• 通讯作者: 8番目)

乳癌骨転移におけるTGF-βシグナルとインビボイメージング

乳腺癌骨转移中的TGF-β信号和体内成像

• 发表时间: 2007
• 作者: Moren A;Imamura T (他3名;2番目);Takeshi Imamura;Takeshi Imamura;今村健志
• 通讯作者: 今村健志