Bio-Protocol (May 2014)

Immunostaining Protocol: P-Smad2 (Xenograft and Mice)

  • Alexandre Calon,
  • Elisa Espinet,
  • Sergio Palomo-Ponce,
  • Daniele Tauriello,
  • Mar Iglesias,
  • María Céspedes,
  • Marta Sevillano,
  • Cristina Nadal,
  • Peter Jung,
  • Xiang Zhang,
  • Daniel Byrom,
  • Antoni Riera,
  • David Rossell,
  • Ramón Mangues,
  • Joan Massague,
  • Elena Sancho,
  • Eduard Batlle

DOI
https://doi.org/10.21769/BioProtoc.1119
Journal volume & issue
Vol. 4, no. 9

Abstract

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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.