Abstract
Purpose
NF-κB transcription factor has been associated with cancer development and chemoresistance. We studied the signaling pathway activated by doxorubicin (DOX) leading to NF-κB activation in breast cancer cells.
Methods
NF-κB activity was evaluated by electrophoretic mobility shift in T47D, ZR75.30 and primary culture (MBCDF) from a ductal infiltrating carcinoma. Cell viability was measured by crystal violet. Western blotting was performed to check the expression and phosphorylation of IκBα Ser-32/36. c-Abl was inhibited with Imatinib or by overexpressing a dominant negative form of c-Abl (K290R).
Results
We found a correlation between sensitivity to DOX and amplitude of NF-κB activation. In cells least sensitive to DOX, NF-κB remained activated for longer time (T47D and MBCDF). The opposite effect was observed in cells sensitive to DOX (ZR75.30). DOX did not induce IκBα degradation or Ser-32/36 phosphorylation. Instead, there were modifications in the levels of IκBα tyrosine phosphorylation, suggesting an atypical NF-κB activation. In DOX-resistant cells, Imatinib treatment reduced IκBα tyrosine phosphorylation and NF-κB activity. The Imatinib–DOX combination significantly enhanced cell death of T47D and MBCDF breast cancer cells. Overexpression of c-Abl K290R in T47D and MBCDF cells reduced basal and DOX-induced NF-κB activation as well as IκBα tyrosine phosphorylation. In c-Abl K290R cells, DOX treatment did not mimic the combination Imatinib–DOX-induced cell death.
Conclusions
Inhibition of c-Abl inactivated IκBα/NF-κB pathway is associated with IκBα tyrosine phosphorylation in breast cancer cells. These results also raise the potential use of a combined therapy with Imatinib and DOX for breast cancer patients.
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Abbreviations
- NF-κB:
-
Nuclear factor kappa-light-chain-enhancer of activated B cells
- IκBα:
-
Nuclear factor of kappa light polypeptide gene enhancer in B cell inhibitor, alpha
- DOX:
-
Doxorubicin
- EMSA:
-
Electrophoretic mobility shift assay
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Acknowledgments
We acknowledge the technical work of Karla Alejandra López Zelada, Julio César Garibay Díaz and Carmita Pérez Guitar. We are grateful to Dr. Adolfo García-Sainz and Dr. Alberto Huberman for their critical review of the manuscript. MJIS and AZD are members of The National System for Researchers (Sistema Nacional de Investigadores, SNI). This work was funded by operating grants to MJIS from CONACyT [102825-3] and AZD from Dirección General del Personal Académico, UNAM (DGAPA) [IN229607-2].
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Esparza-López, J., Medina-Franco, H., Escobar-Arriaga, E. et al. Doxorubicin induces atypical NF-κB activation through c-Abl kinase activity in breast cancer cells. J Cancer Res Clin Oncol 139, 1625–1635 (2013). https://doi.org/10.1007/s00432-013-1476-3
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DOI: https://doi.org/10.1007/s00432-013-1476-3