Tumor Biology

, Volume 37, Issue 12, pp 15825–15833 | Cite as

Notch3 negatively regulates chemoresistance in breast cancers

  • Xiaoting Gu
  • Chunxiao Lu
  • Dongxu He
  • Yangfan Lu
  • Jian Jin
  • Dequan Liu
  • Xin Ma
Original Article


To define the role of the NOTCH signaling pathway in the development of chemoresistance and the associated epithelial–mesenchymal transition (EMT), we investigated the effect of Notch3 on adriamycin (ADM)-resistant human breast cancer cells (MCF-7/ADM cells). We found that Notch3 was downregulated and involved in the chemoresistance of MCF-7/ADM cells, while forced expression of Notch3 reversed the chemoresistance. Furthermore, fos-related antigen 1 (Fra1) was negatively regulated by Notch3 and was highly expressed in MCF-7/ADM cells. Increased Fra1 activated the EMT process. Finally, Notch3 expression was confirmed in clinically chemoresistant samples of breast cancers from patients receiving anthracycline-based chemotherapy. Low expression of Notch3 was an unfavorable predictor of distant relapse-free survival in ER positive breast cancers. Taken together, our findings demonstrate that the Notch3-Fra1 signaling pathway mediates chemoresistance via the EMT.


Notch3 Fra1 EMT Chemoresistance Breast cancer 



This work was supported by the China National Natural Science Foundation 81572940 and 91439131 to Xin Ma; 31200126 and 31550006 to Dongxu He; 81272358 to Feng Gu; the Natural Science Foundation for Distinguished Young Scholars of Jiangsu Province (BK20140004 to Xin Ma); the Program for New Century Excellent Talents in University of The Ministry of Education of China (NCET-12-0880) to Xin Ma; the National High Technology Research and Development Program (863 Program) of China (SQ2015AA020948) to Xin Ma; and the Fundamental Research Funds for the Central Universities (JUSRP51311A, JUSRP51615B and JUSRP51519) to Xin Ma, Dongxu He and Jin Jian. We thank Dr. IC Bruce for reading the manuscript.

Author contributions

Authors who participated in research design are as follows: X.-T. Gu, D.-X. He, X. Ma, and D-Q. Liu. Authors who conducted experiments are as follows: X.-T. Gu, C.-X. Lu, and Y.-F. Lu. Authors who performed data analysis are as follows: X.-T. Gu and D.-X. He. Authors who wrote or contributed to the writing of the manuscript are as follows: X.-T. Gu and D.-X. He.

Compliance with ethical standards

Conflicts of interest


Supplementary material

13277_2016_5412_Fig6_ESM.gif (144 kb)
Fig. S1

Notch3-Fra1 signaling pathway in MCF-7/PTX. (a) Notch3 expression was lower in MCF-7/PTX (PTX) cells than in MCF-7/WT (WT) cells when analyzed by quantitative real-time PCR (**P < 0.001 versus MCF-7/WT). (b) Notch3 expression was decreased in MCF-7/PTX cells as analyzed by western blotting (∼280 kDa, ***P < 0.0001 versus MCF-7/WT cells). (c) Expression of Fra1 was increased in MCF-7/PTX cells versus MCF-7/WT cells when analyzed by quantitative real-time PCR (***P < 0.0001 versus MCF-7/WT cells). (d) Fra1 expression was decreased in MCF-7/PTX + N3CEG cells as analyzed by western blotting (∼29 kDa, *P < 0.05 versus MCF-7/PTX + pCEG cells). (e) Expression and location of E-cadherin and vimentin in MCF-7/PTX + N3CEG (Notch3 overexpressed in MCF-7/PTX), MCF-7/PTX + pCEG (MCF-7/PTX transfected by an empty plasmid), MCF-7/PTX + siFra1 (Fra1 knocked down in MCF-7/WT), and MCF-7/PTX + NC (MCF-7/PTX transfected by a scrambled siRNA) cells analyzed by confocal microscopy. (b) Cell migration with different treatments (***P < 0.0001 PTX + N3CEG vs PTX + pCEG, ***P < 0.0001 PTX + siFra1 vs PTX + NC). (GIF 143 kb)

13277_2016_5412_MOESM1_ESM.tif (1.6 mb)
High Resolution Image (TIFF 1654 kb)


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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2016

Authors and Affiliations

  1. 1.School of Pharmaceutical SciencesJiangnan UniversityWuxiChina
  2. 2.National Engineering Laboratory for Cereal Fermentation TechnologyJiangnan UniversityWuxiChina
  3. 3.Department of Breast Surgery, The Third Affiliated HospitalKunming Medical UniversityKunmingChina

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