Abrogation of TGF-β signaling enhances chemokine production and correlates with prognosis in human breast cancer
Brian Bierie, … , Yu Shyr, Harold L. Moses
Brian Bierie, … , Yu Shyr, Harold L. Moses
Published May 18, 2009
Citation Information: J Clin Invest. 2009;119(6):1571-1582. https://doi.org/10.1172/JCI37480.
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Research Article Oncology

Abrogation of TGF-β signaling enhances chemokine production and correlates with prognosis in human breast cancer

Abstract

In human breast cancer, loss of carcinoma cell–specific response to TGF-β signaling has been linked to poor patient prognosis. However, the mechanisms through which TGF-β regulates these processes remain largely unknown. In an effort to address this issue, we have now identified gene expression signatures associated with the TGF-β signaling pathway in human mammary carcinoma cells. The results strongly suggest that TGF-β signaling mediates intrinsic, stromal-epithelial, and host-tumor interactions during breast cancer progression, at least in part, by regulating basal and oncostatin M–induced CXCL1, CXCL5, and CCL20 chemokine expression. To determine the clinical relevance of our results, we queried our TGF-β–associated gene expression signatures in 4 human breast cancer data sets containing a total of 1,319 gene expression profiles and associated clinical outcome data. The signature representing complete abrogation of TGF-β signaling correlated with reduced relapse-free survival in all patients; however, the strongest association was observed in patients with estrogen receptor–positive (ER-positive) tumors, specifically within the luminal A subtype. Together, the results suggest that assessment of TGF-β signaling pathway status may further stratify the prognosis of ER-positive patients and provide novel therapeutic approaches in the management of breast cancer.

Authors

Brian Bierie, Christine H. Chung, Joel S. Parker, Daniel G. Stover, Nikki Cheng, Anna Chytil, Mary Aakre, Yu Shyr, Harold L. Moses

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Figure 1

Recombination of TβRII and induction of TGF-β–dependent EMT in MMTV-PyVmT mammary carcinoma cells.

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Recombination of TβRII and induction of TGF-β–dependent EMT in MMTV-PyVm...
(A) Analysis of Southern blot hybridization demonstrated that the independently derived polyclonal TβRII(fl/fl;PY) control mammary carcinoma cells (FL1, FL2, and FL3; biological replicates) had intact floxed Tgfbr2 alleles, with no evidence of recombination. Alternatively, the independently derived polyclonal TβRII(WKO;PY) carcinoma cells (KO1, KO2, and KO3; biological replicates) were completely recombined, with no evidence of a remaining floxed Tgfbr2 allele. (B) The growth of TβRII(fl/fl;PY) control carcinoma cell lines, measured by tritiated thymidine incorporation 24 hours after stimulation, was markedly inhibited by TGF-β stimulation, whereas the TβRII(WKO;PY) carcinoma cell growth was not altered. Results represent median transformed mean values ± SEM. (C and D) In response to TGF-β stimulation (10 ng/ml) for 48 hours, the TβRII(fl/fl;PY) control carcinoma cells demonstrated consistent changes in morphology and cell scattering that suggested that EMT had occurred (original magnification, ×10). (E–H) Loss of E-cadherin (E and F) and changes in F-actin localization from the cell membrane (G) to predominant association with stress fibers (H) further confirmed an EMT-like state in TβRII(fl/fl;PY) cells that had been treated with 10 ng/ml of TGF-β for 48 hours (E–H; original magnification, ×40). (I and J) Loss of TGF-β signaling in TβRII(WKO;PY) cells did not result in a spontaneous state of EMT, as determined by the presence of membrane-bound E-cadherin, in the presence or absence of TGF-β ligand at 10 ng/ml after 48 hours of stimulation (original magnification, ×40).