Abstract
The transcription factor nuclear factor kappa B (NF-κB) is constitutively active in both cancer cells and stromal cells of breast cancer; however, the precise role of activated NF-κB in cancer progression is not known. Using parental MCF10A cells and a variant that expresses the myoepithelial marker p63 stably overexpressing the constitutively active p65 subunit of NF-κB (MCF10A/p65), we show that NF-κB suppresses the expression of epithelial specific genes E-cadherin and desmoplakin and induces the expression of the mesenchymal specific gene vimentin. P65 also suppressed the expression of p63 and the putative breast epithelial progenitor marker cytokeratin 5/6. MCF10A/p65 cells were phenotypically similar to cells undergoing epithelial to mesenchymal transition (EMT). MCF10A/p65 cells failed to form characteristic acini in three-dimensional Matrigel. Analysis of parental and MCF10A/p65 cells for genes previously shown to be involved in EMT revealed elevated expression of ZEB-1 and ZEB-2 in MCF10A/p65 cells compared to parental cells. In transient transfection assays, p65 increased ZEB-1 promoter activity. Furthermore, MCF10A cells overexpressing ZEB-1 showed reduced E-cadherin and p63 expression and displayed an EMT phenotype. The siRNA against ZEB-1 or ZEB-2 reduced the number of viable MCF10A/p65 but not parental cells, suggesting the dependence of MCF10A/p65 cells to ZEB-1 and ZEB-2 for cell cycle progression or survival. MCF10A cells chronically exposed to tumor necrosis factor alpha (TNFα), a potent NF-κB inducer, also exhibited the EMT-like phenotype and ZEB-1/ZEB-2 induction, both of which were reversed following TNFα withdrawal.
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Abbreviations
- EMT:
-
epithelial to mesenchymal transition
- IL-1:
-
interleukin-1
- MnSOD:
-
manganese superoxide dismutase
- NF-κB:
-
nuclear factor kappaB
- NLS:
-
nuclear localization signal
- TGFβ:
-
transforming growth factor beta
- TNF:
-
tumor necrosis factor
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Acknowledgements
We thank Meei-Huey Jeng and George Sledge for providing the MCF10A(E) and MCF10A(M) cells, respectively, Tom Genetta for the ZEB-1 cDNA, Geert Berx for the ZEB-2 cDNA, and Louis Pelus for p63 cDNA. We are also grateful to Drs Attaya Suvannasankha, Zhuo Wang and Colin Crean for their critical review of our manuscript. HN is Marian J Morrison Investigator for Breast Cancer Research. This work is supported by the Department of Defense Grant DAMD17-01-1-0274 to H.N.
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Chua, H., Bhat-Nakshatri, P., Clare, S. et al. NF-κB represses E-cadherin expression and enhances epithelial to mesenchymal transition of mammary epithelial cells: potential involvement of ZEB-1 and ZEB-2. Oncogene 26, 711–724 (2007). https://doi.org/10.1038/sj.onc.1209808
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DOI: https://doi.org/10.1038/sj.onc.1209808
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