Lack of transforming growth factor-β signaling promotes collective cancer cell invasion through tumor-stromal crosstalk.
Lack of transforming growth factor-β signaling promotes collective cancer cell invasion through tumor-stromal crosstalk.
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DOI：
10.1186/bcr3217
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发表时间：
2012-07-02
期刊：
影响因子：
--
通讯作者：
Moses HL
中科院分区：
文献类型：
--
作者：
Matise LA;Palmer TD;Ashby WJ;Nashabi A;Chytil A;Aakre M;Pickup MW;Gorska AE;Zijlstra A;Moses HL
转化生长因子β（TGF-β）在肿瘤进展过程中具有双重作用，最初作为抑制剂，然后作为促进剂。上皮TGF-β信号转导调节成纤维细胞募集和活化。同时，间质成纤维细胞中的TGF-β信号传导抑制邻近上皮中的肿瘤发生，而其消融增强肿瘤形成。关于TGF-β信号传导对肿瘤发生的贡献已知之甚多，但对TGF-β在肿瘤进展期间上皮-间质迁移中的作用知之甚少。我们假设TGF-β是促进乳腺肿瘤细胞迁移和侵袭的肿瘤-基质相互作用的关键调节因子。将从MMTV-PyVmT转化生长因子-β受体II敲除（TβRII KO）或TβRIIfl/fl对照小鼠中分离的荧光标记的小鼠乳腺癌细胞与乳腺成纤维细胞结合，并异种移植到鸡胚绒毛尿囊膜上。这些组合异种移植物被用作研究上皮-基质串扰的模型。迁移的活体成像监测卵外，并在卵内转移进行了研究。通过激光捕获显微切割分离来自卵内肿瘤的上皮RNA，并分析以鉴定响应于TGF-β信号传导损失的基因表达变化。异种移植物的活体显微镜检查显示，乳腺成纤维细胞促进依赖于上皮TGF-β信号传导的两种迁移表型：单细胞/链迁移或集体迁移。在上皮-间质边界，TβRIIfl/fl癌细胞的单细胞/链迁移的特征在于α-平滑肌肌动蛋白和波形蛋白的表达，而TβRII KO癌细胞的集体迁移通过E-cadherin+/p120+/β-catenin+簇来鉴定。TβRII KO肿瘤也表现出比TβRIIfl/fl肿瘤高两倍的转移，归因于增强的外渗能力。与TβRIIfl/fl上皮相比，在TβRII KO肿瘤上皮中，Igfbp 4和Tspan 13表达上调，而Col 1 α2、Bmp 7、Gng 11、Vcan、Tmeff 1和Dsc 2表达下调。对培养细胞的免疫印迹和定量PCR分析验证了这些靶点，并将Tmeff 1表达与TGF-β不敏感乳腺癌的疾病进展相关。成纤维细胞刺激的癌细胞利用TGF-β信号传导来驱动单个细胞/链迁移，但在TGF-β信号传导不存在的情况下集体迁移。这些迁移模式涉及几种上皮细胞向间充质细胞转化途径的信号调节。我们关于上皮-基质相互作用中TGF-β信号传导的发现对于确定可作为乳腺癌治疗手段的迁移机制非常重要。
Transforming growth factor beta (TGF-β) has a dual role during tumor progression, initially as a suppressor and then as a promoter. Epithelial TGF-β signaling regulates fibroblast recruitment and activation. Concurrently, TGF-β signaling in stromal fibroblasts suppresses tumorigenesis in adjacent epithelia, while its ablation potentiates tumor formation. Much is known about the contribution of TGF-β signaling to tumorigenesis, yet the role of TGF-β in epithelial-stromal migration during tumor progression is poorly understood. We hypothesize that TGF-β is a critical regulator of tumor-stromal interactions that promote mammary tumor cell migration and invasion. Fluorescently labeled murine mammary carcinoma cells, isolated from either MMTV-PyVmT transforming growth factor-beta receptor II knockout (TβRII KO) or TβRIIfl/fl control mice, were combined with mammary fibroblasts and xenografted onto the chicken embryo chorioallantoic membrane. These combinatorial xenografts were used as a model to study epithelial-stromal crosstalk. Intravital imaging of migration was monitored ex ovo, and metastasis was investigated in ovo. Epithelial RNA from in ovo tumors was isolated by laser capture microdissection and analyzed to identify gene expression changes in response to TGF-β signaling loss. Intravital microscopy of xenografts revealed that mammary fibroblasts promoted two migratory phenotypes dependent on epithelial TGF-β signaling: single cell/strand migration or collective migration. At epithelial-stromal boundaries, single cell/strand migration of TβRIIfl/fl carcinoma cells was characterized by expression of α-smooth muscle actin and vimentin, while collective migration of TβRII KO carcinoma cells was identified by E-cadherin+/p120+/β-catenin+ clusters. TβRII KO tumors also exhibited a twofold greater metastasis than TβRIIfl/fl tumors, attributed to enhanced extravasation ability. In TβRII KO tumor epithelium compared with TβRIIfl/fl epithelium, Igfbp4 and Tspan13 expression was upregulated while Col1α2, Bmp7, Gng11, Vcan, Tmeff1, and Dsc2 expression was downregulated. Immunoblotting and quantitative PCR analyses on cultured cells validated these targets and correlated Tmeff1 expression with disease progression of TGF-β-insensitive mammary cancer. Fibroblast-stimulated carcinoma cells utilize TGF-β signaling to drive single cell/strand migration but migrate collectively in the absence of TGF-β signaling. These migration patterns involve the signaling regulation of several epithelial-to-mesenchymal transition pathways. Our findings concerning TGF-β signaling in epithelial-stromal interactions are important in identifying migratory mechanisms that can be targeted as recourse for breast cancer treatment.
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影响因子：
6
作者：
Gorska, AE;Jensen, RA;Moses, HL
通讯作者：
Moses, HL
影响因子：
21.3
作者：
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影响因子：
11.2
作者：
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通讯作者：
Moses, Harold L.
影响因子：
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作者：
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影响因子：
11.2
作者：
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通讯作者：
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