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
Affiliations
Alexandre Calon
Oncology Programme, Institute for Research in Biomedicine, Barcelona, Spain
Elisa Espinet
Oncology Programme, Institute for Research in Biomedicine, Barcelona, Spain
Sergio Palomo-Ponce
Oncology Programme, Institute for Research in Biomedicine, Barcelona, Spain
Daniele Tauriello
Oncology Programme, Institute for Research in Biomedicine, Barcelona, Spain
Mar Iglesias
Department of Pathology, Hospital Universitari del Mar, Barcelona, Spain
María Céspedes
IIB Sant Pau, Hospital de la Santa Creu i Sant Pau, Barcelona, Spain
Marta Sevillano
Oncology Programme, Institute for Research in Biomedicine, Barcelona, Spain
Cristina Nadal
Institut de Malalties Hemato-Oncològiques, Hospital Clínic, Barcelona, Spain
Peter Jung
Oncology Programme, Institute for Research in Biomedicine, Barcelona, Spain
Xiang Zhang
Cancer Biology and Genetics Program, Memorial Sloan-Kettering Cancer Center, New York, USA
Daniel Byrom
Chemistry and Molecular Pharmacology Programme, Institute for Research in Biomedicine, Barcelona, Spain
Antoni Riera
Chemistry and Molecular Pharmacology Programme, Institute for Research in Biomedicine, Barcelona, Spain
David Rossell
Biostatistics and Bioinformatics Unit, Institute for Research in Biomedicine, Barcelona, Spain
Ramón Mangues
IIB Sant Pau, Hospital de la Santa Creu i Sant Pau, Barcelona, Spain
Joan Massague
Cancer Biology and Genetics Program, Memorial Sloan-Kettering Cancer Center, New York, USA
Elena Sancho
Oncology Programme, Institute for Research in Biomedicine, Barcelona, Spain
Eduard Batlle
Oncology Programme, Institute for Research in Biomedicine, Barcelona, Spain
DOI
https://doi.org/10.21769/BioProtoc.1119
Journal volume & issue
Vol. 4,
no. 9
Abstract
Read online
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.
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