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），是调节广泛的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信号促进晚期癌症中的肿瘤生长和转移。TGF-β信号通路也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，我们研究了一种新型TGF-β I型受体（TBR-I）激酶抑制剂Ki 26894对MDA-231-D细胞骨转移的影响。Ki 26894阻断了MDA-231-D细胞中的TGF-β信号传导，如通过抑制Smad 2的磷酸化和抑制TGF-β反应性报告基因活性所检测到的。此外，Ki ...更多信息 26894在体外可降低TGF-β诱导的MDA-231-D细胞的运动性和侵袭性。Ki 26894还抑制TGF-β刺激的MDA-231-D细胞的纤溶酶原激活物抑制剂-1（派-1）、甲状旁腺相关蛋白（PTHrP）和白细胞介素-11（IL-11）mRNA的转录。X射线照相术显示，在将MDA-231-D细胞接种到BALB/c nu/nu雌性小鼠的左心室中之前1天开始的系统性Ki 26894治疗导致乳腺癌细胞的骨转移减少。此外，与溶剂处理的小鼠相比，Ki 26894延长了接种MDA-231-D细胞的小鼠的存活时间。这些发现表明TβR-I激酶抑制剂如Ki 26894可能有助于阻断晚期癌症的进展。此外，我们鉴定了软骨细胞中差异表达的DEC 1（也称为SHARP 2和Stral 3）作为TGF-β信号传导的下游靶点，其促进乳腺癌细胞的存活。在小鼠乳腺癌细胞系JygMC（A）和4 T1中，TBR-β激酶抑制剂A-44-03和SB 431542在无血清条件下诱导细胞凋亡。寡核苷酸芯片和实时逆转录PCR分析显示，TGF-β可诱导这些细胞中的DEC 1，而DEC 1的增加可被TβR-I激酶抑制剂以及显性阴性TGF-β II型受体的表达所抑制。过表达DEC 1可抑制A-44-03诱导的JygMC（A）细胞凋亡，敲低内源性DEC 1可抑制TGF-β促进的细胞存活。此外，DEC 1的显性负突变体在体内阻止JygMC（A）细胞的肺和肝转移。因此，我们的观察结果提供了新的见解TGF-β介导的细胞存活和转移的癌症的分子机制。少

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

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

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.