Protein expression changes during human triple negative breast cancer cell line progression to lymph node metastasis in a xenografted model in nude mice

作者：Paula Roberti Maria; Martin Arriaga Juan; Bianchini Michele; Ramiro Quinta Hector; Ines Bravo Alicia; Mariel Levy Estrella; Mordoh Jose; Marcela Barrio Maria

来源：Cancer Biology and Therapy, 2012, 13(11): 1123-1140.

DOI：10.4161/cbt.21187

• triple negative breast cancer
• metastasis
• protein profile
• angiogenesis

Triple negative breast cancers (TNBC) lacking hormone receptors and HER-2 amplification are very aggressive tumors. Since relevant differences between primary tumors and metastases could arise during tumor progression as evidenced by phenotypic discordances reported for hormonal receptors or HER-2 expression, in this analysis we studied changes that occurred in our TNBC model IIB-BR-G throughout the development of IIB-BR-G-(MTS6) metastasis to the lymph nodes (LN) in nude mice, using an antibody-based protein array to characterize their expression profile. We also analyzed their growth kinetics, migration, invasiveness and cytoskeleton structure in vitro and in vivo.
In vitro IIB-BR-G-(MTS6) cells grew slower but showed higher anchorage independent growth. In vivo IIB-BR-G-(MTS6) tumors grew significantly faster and showed a 100% incidence of LN metastasis after s.c. inoculation, although no metastasis was observed for IIB-BR-G. CCL3, IL1 beta, CXCL1, CSF2, CSF3, IGFBP1, IL1 alpha, IL6, IL8, CCL20, PLAUR, PlGF and VEGF were strongly upregulated in IIB-BR-G-(MTS6) while CCL4, ICAM3, CXCL12, TNFRSF18, FIGF were the most downregulated proteins in the metastatic cell line. IIB-BR-G-(MTS6) protein expression profile could reflect a higher NF kappa B activation in these cells. In vitro, IIB-BR-G displayed higher migration but IIB-BR-G-(MTS6) had more elevated matrigel invasion ability. In agreement with that observation, IIB-BR-G-(MTS6) had an upregulated expression of MMP1, MMP9, MMP13, PLAUR and HGF.IIB-BR-G-(MTS6) tumors presented also higher local lymphatic invasion than IIB-BR-G but similar lymphatic vessel densities. VEGFC and VEGFA/B expression were higher both in vitro and in vivo for IIB-BR-G-(MTS6). IIB-BR-G-(MTS6) expressed more vimentin than IB-BR-G cells, which was mainly localized in the cellular extremities and both cell lines are E-cadherin negative.
Our results suggest that IIB-BR-G-(MTS6) cells have acquired a pronounced epithelial-to-mesenchymal transition phenotype. Protein expression changes observed between primary tumor-derived IIB-BR-G and metastatic IIB-BR-G-(MTS6) TNBC cells suggest potential targets involved in the control of metastasis.

• 出版日期2012-9