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、迁移和凋亡。 TGF-β信号传导促进晚期癌症的肿瘤生长和转移。 TGF-β信号通路相应地成为肿瘤学领域药物开发的一个有吸引力的靶点。我们研究了TGF-β和BMP信号通路在骨转移过程中的作用。我们首先建立了人乳腺癌 MDA-MB-231 细胞的高度骨转移变体，称为 MDA-MB-231-5a-D (MDA-231-D)。接下来，我们研究了新型 TGF-β I 型受体 (TBR-I) 激酶抑制剂 Ki26894 对 MDA-231-D 细胞骨转移的影响。 Ki26894 阻断 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 响应报告基因活性来检测。此外，Ki … 更多 26894 降低了体外 TGF-β 诱导的 MDA-231-D 细胞的运动和侵袭。 Ki26894 还抑制受 TGF-β 刺激的 MDA-231-D 细胞的纤溶酶原激活剂抑制剂-1 (PAI-1)、甲状旁腺激素相关蛋白 (PTHrP) 和白细胞介素 11 (IL-11) mRNA 的转录。 X射线照相显示，在将MDA-231-D细胞接种到BALB/c nu/nu雌性小鼠的左心室前1天开始全身Ki26894治疗，导致乳腺癌细胞的骨转移减少。此外，与媒介物处理的小鼠相比，Ki26894 延长了接种 MDA-231-D 细胞的小鼠的存活时间。这些发现表明，Ki26894等TβR-I激酶抑制剂可能有助于阻止晚期癌症的进展。此外，我们发现软骨细胞1（DEC1，也称为SHARP2和Stral3）中差异表达的蛋白是TGF-β信号传导的下游靶标，可促进乳腺癌细胞的存活。在小鼠乳腺癌细胞系 JygMC (A) 和 4T1 中，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，并且TβR-I激酶抑制剂以及显性失活TGF-β II型受体的表达抑制DEC1的增加。 DEC1 的过度表达可阻止 A-44-03 诱导的 JygMC (A) 细胞凋亡，而内源性 DEC1 的敲除可消除 TGF-β 促进的细胞存活。此外，DEC1 的显性失活突变体可阻止 JygMC (A) 细胞体内的肺和肝转移。因此，我们的观察结果为控制 TGF-β 介导的细胞存活和癌症转移的分子机制提供了新的见解。较少的

项目成果

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

癌研究におけるインビボバイオイメージングの応用

体内生物成像在癌症研究中的应用

• 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
• 通讯作者: 今村健志

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

Modulation of the functional binding sites for TGF-β on the type II receptor leads to suppression of TGF-β signaling

• DOI: 10.1038/sj.onc.1210123
• 发表时间: 2007-05-17
• 期刊: ONCOGENE
• 影响因子: 8
• 作者: Shimanuki, T.;Hara, T.;Miyazono, K.
• 通讯作者: Miyazono, K.

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