| Immunostaining Protocol: P-Smad2 (Xenograft and Mice)
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Author:
Alexandre Calon, Elisa Espinet, Sergio Palomo-Ponce, Daniele V. F. Tauriello, Mar Iglesias, María Virtudes Céspedes, Marta Sevillano, Cristina Nadal, Peter Jung, Xiang H.-F. Zhang, Daniel Byrom, Antoni Riera, David Rossell, Ramón Mangues, Joan Massague, Elena Sancho and Eduard Batlle,
Date:
2014-05-05
[Abstract] Metastasis depends on a gene program expressed by the tumor microenvironment upon TGF-beta stimulation. CRC (Colorectal cancer) cell lines did not induce robust stromal TGF-beta responses when injected into nude mice as shown by lack of p-SMAD2 accumulation in tumor-associated stromal cells. To enforce high TGF-beta signaling in xenografts, we engineered CRC cell lines to secrete active TGF-beta. Subcutaneous tumors obtained from HT29-M6TGF-β, KM12L4aTGF-β cells and SW48TGF-β cells contained abundant p-SMAD2+ stromal cells.
[摘要] 转移依赖于在TGF-β刺激时由肿瘤微环境表达的基因程序。 当注射到裸鼠中时，CRC（结肠直肠癌）细胞系不诱导强烈的基质TGF-β应答，如肿瘤相关基质细胞中缺乏p-SMAD2积累所示。 为了在异种移植物中强化高TGF-β信号传导，我们设计CRC细胞系以分泌活性TGF-β。 从HT29-M6TGF-β，KM12L4aTGF-β细胞和SW48TGF-β细胞获得的皮下肿瘤含有丰富的p-SMAD2 + 基质细胞。
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| Immunostaining Protocol: P-Stat3 (Xenograft and Mice)
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Author:
Alexandre Calon, Elisa Espinet, Sergio Palomo-Ponce, Daniele V. F. Tauriello, Mar Iglesias, María Virtudes Céspedes, Marta Sevillano, Cristina Nadal, Peter Jung, Xiang H.-F. Zhang, Daniel Byrom, Antoni Riera, David Rossell, Ramón Mangues, Joan Massague, Elena Sancho and Eduard Batlle,
Date:
2014-05-05
[Abstract] We sought to understand the mechanisms behind the potent effect of stromal TGF-beta program on the capacity of colorectal cancer (CRC) cells to initiate metastasis. We discovered that mice subcutaneous tumors and metastases generated in the context of a TGF-beta activated microenvironment displayed prominent accumulation of p-STAT3 in CRC cells compared with those derived from control cells. STAT3 signaling depended on GP130 as shown by strong reduction of epithelial p STAT3 levels upon GP130 shRNA-mediated knockdown in CRC cells.
[摘要] 我们试图了解基质TGF-β程序对结肠直肠癌（CRC）细胞启动转移能力的有效作用背后的机制。 我们发现小鼠皮下肿瘤和转移生成的TGF-β激活微环境的情况下显示显着积累的p-STAT3在CRC细胞相比那些衍生自控制细胞。 STAT3信号传导依赖于GP130，如通过GP130shRNA介导的CRC细胞敲除的上皮p STAT3水平的强烈减少所示。
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