Article | Published:

Translational Therapeutics

Dasatinib sensitises triple negative breast cancer cells to chemotherapy by targeting breast cancer stem cells

British Journal of Cancervolume 119pages14951507 (2018) | Download Citation

Abstract

Background

Patients with triple negative breast cancer (TNBC) exhibit poor prognosis and are at high risk of tumour relapse, due to the resistance to chemotherapy. These aggressive phenotypes are in part attributed to the presence of breast cancer stem cells (BCSCs). Therefore, targeting BCSCs is a priority to overcoming chemotherapy failure in TNBCs.

Methods

We generated paclitaxel (pac)-resistant TNBC cells which displayed higher sphere forming potential and percentage of BCSC subpopulations compared to the parental cells. A screen with various kinase inhibitors revealed dasatinib, a Src kinase family inhibitor, as a potent suppressor of BCSC expansion/sphere formation in pac-resistant TNBC cells.

Results

We found dasatinib to block pac-induced BCSC enrichment and Src activation in both parental and pac-resistant TNBC cells. Interestingly, dasatinib induced an epithelial differentiation of the pac-resistant mesenchymal cells, resulting in their enhanced sensitivity to paclitaxel. The combination treatment of dasatinib and paclitaxel not only decreased the BCSCs numbers and their sphere forming capacity but also synergistically reduced cell viability of pac-resistant cells. Preclinical models of breast cancer further demonstrated the efficiency of the dasatinib/paclitaxel combination treatment in inhibiting tumour growth.

Conclusions

Dasatinib is a promising anti-BCSC drug that could be used in combination with paclitaxel to overcome chemoresistance in TNBC.

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Additional information

The study was performed in accordance with the Declaration of Helsinki.

References

  1. 1.

    Schneider, B. P. et al. Triple-negative breast cancer: risk factors to potential targets. Clin. Cancer Res. 14, 8010–8018 (2008).

  2. 2.

    Andre, F. & Zielinski, C. C. Optimal strategies for the treatment of metastatic triple-negative breast cancer with currently approved agents. Ann. Oncol. 23, vi46–51 (2012).

  3. 3.

    Magee, J. A., Piskounova, E. & Morrison, S. J. Cancer stem cells: impact, heterogeneity, and uncertainty. Cancer Cell 21, 283–296 (2012).

  4. 4.

    Wicha, M. S., Liu, S. & Dontu, G. Cancer stem cells: an old idea—a paradigm shift. Cancer Res. 66, 1883–1890 (2006).

  5. 5.

    Al-Hajj, M., Wicha, M. S., Benito-Hernandez, A., Morrison, S. J. & Clarke, M. F. Prospective identification of tumorigenic breast cancer cells. Proc. Natl Acad. Sci. USA 100, 3983–3988 (2003).

  6. 6.

    Ginestier, C. et al. ALDH1 is a marker of normal and malignant human mammary stem cells and a predictor of poor clinical outcome. Cell Stem Cell 1, 555–567 (2007).

  7. 7.

    Lombardo, Y. et al. Bone morphogenetic protein 4 induces differentiation of colorectal cancer stem cells and increases their response to chemotherapy in mice. Gastroenterology 140, 297–309 (2011).

  8. 8.

    Wielenga, M. C. B. et al. ER-stress-induced differentiation sensitizes colon cancer stem cells to chemotherapy. Cell Rep. 13, 489–494 (2015).

  9. 9.

    Campos, B. et al. Differentiation therapy exerts antitumor effects on stem-like glioma cells. Clin. Cancer Res. 16, 2715–2728 (2010).

  10. 10.

    Yeatman, T. J. A renaissance for SRC. Nat. Rev. Cancer 4, 470–480 (2004).

  11. 11.

    Wheeler, D. L., Iida, M. & Dunn, E. F. The role of Src in solid tumors. Oncologist 14, 667–678 (2009).

  12. 12.

    Ahluwalia, M. S., de Groot, J., Liu, W. M. & Gladson, C. L. Targeting SRC in glioblastoma tumors and brain metastases: rationale and preclinical studies. Cancer Lett. 298, 139–149 (2010).

  13. 13.

    Summy, J. M. & Gallick, G. E. Src family kinases in tumor progression and metastasis. Cancer Metastas-. Rev. 22, 337–358 (2003).

  14. 14.

    Zhang, X. H. et al. Latent bone metastasis in breast cancer tied to Src-dependent survival signals. Cancer Cell 16, 67–78 (2009).

  15. 15.

    Thakur, R., Trivedi, R., Rastogi, N., Singh, M. & Mishra, D. P. Inhibition of STAT3, FAK and Src mediated signaling reduces cancer stem cell load, tumorigenic potential and metastasis in breast cancer. Sci. Rep. 5, 10194 (2015).

  16. 16.

    Steinberg, M. Dasatinib: a tyrosine kinase inhibitor for the treatment of chronic myelogenous leukemia and philadelphia chromosome-positive acute lymphoblastic leukemia. Clin. Ther. 29, 2289–2308 (2007).

  17. 17.

    Lehmann, B. D. et al. Identification of human triple-negative breast cancer subtypes and preclinical models for selection of targeted therapies. J. Clin. Invest. 121, 2750–2767 (2011).

  18. 18.

    Kurebayashi, J. et al. Preferential antitumor effect of the Src inhibitor dasatinib associated with a decreased proportion of aldehyde dehydrogenase 1-positive cells in breast cancer cells of the basal B subtype. BMC Cancer 10, 568 (2010).

  19. 19.

    Tian, J. et al. Cyclooxygenase-2 regulates TGFbeta-induced cancer stemness in triple-negative breast cancer. Sci. Rep. 7, 40258 (2017).

  20. 20.

    Dai, M. et al. CDK4 regulates cancer stemness and is a novel therapeutic target for triple-negative breast cancer. Sci. Rep. 6, 35383 (2016).

  21. 21.

    Charafe-Jauffret, E. et al. Breast cancer cell lines contain functional cancer stem cells with metastatic capacity and a distinct molecular signature. Cancer Res. 69, 1302–1313 (2009).

  22. 22.

    Fillmore, C. M. & Kuperwasser, C. Human breast cancer cell lines contain stem-like cells that self-renew, give rise to phenotypically diverse progeny and survive chemotherapy. Breast Cancer Res. 10, R25 (2008).

  23. 23.

    Bhola, N. E. et al. TGF-beta inhibition enhances chemotherapy action against triple-negative breast cancer. J. Clin. Invest. 123, 1348–1358 (2013).

  24. 24.

    Ponti, D. et al. Isolation and in vitro propagation of tumorigenic breast cancer cells with stem/progenitor cell properties. Cancer Res. 65, 5506–5511 (2005).

  25. 25.

    Iorns, E. et al. A new mouse model for the study of human breast cancer metastasis. PLoS ONE 7, e47995 (2012).

  26. 26.

    Massihnia, D. et al. Triple negative breast cancer: shedding light onto the role of pi3k/akt/mtor pathway. Oncotarget 7, 60712–60722 (2016).

  27. 27.

    Lebrun, J.-J. The dual role of TGFβ in human cancer: from tumor suppression to cancer metastasis. ISRN Mol. Biol. 2012, 28 (2012).

  28. 28.

    Crown, J., O’Shaughnessy, J. & Gullo, G. Emerging targeted therapies in triple-negative breast cancer. Ann. Oncol. 23, vi56–65 (2012).

  29. 29.

    Kalimutho, M. et al. Targeted therapies for triple-negative breast cancer: combating a stubborn disease. Trends Pharmacol. Sci. 36, 822–846 (2015).

  30. 30.

    Xie, X. et al. c-Jun N-terminal kinase promotes stem cell phenotype in triple-negative breast cancer through upregulation of Notch1 via activation of c-Jun. Oncogene 36, 2599–2608 (2017).

  31. 31.

    Xia, P. & Xu, X. Y. PI3K/Akt/mTOR signaling pathway in cancer stem cells: from basic research to clinical application. Am. J. Cancer Res. 5, 1602–1609 (2015).

  32. 32.

    Cheng, J. et al. MEK1 signaling promotes self-renewal and tumorigenicity of liver cancer stem cells via maintaining SIRT1 protein stabilization. Oncotarget 7, 20597–20611 (2016).

  33. 33.

    Chen, F. JNK-induced apoptosis, compensatory growth, and cancer stem cells. Cancer Res. 72, 379–386 (2012).

  34. 34.

    Abhold, E. L. et al. EGFR kinase promotes acquisition of stem cell-like properties: a potential therapeutic target in head and neck squamous cell carcinoma stem cells. PLoS ONE 7, e32459 (2012).

  35. 35.

    Mani, S. A. et al. The epithelial-mesenchymal transition generates cells with properties of stem cells. Cell 133, 704–715 (2008).

  36. 36.

    Edmondson, R., Broglie, J. J., Adcock, A. F. & Yang, L. Three-dimensional cell culture systems and their applications in drug discovery and cell-based biosensors. Assay. Drug Dev. Technol. 12, 207–218 (2014).

  37. 37.

    Liu, F. et al. Prolactin/Jak2 directs apical/basal polarization and luminal linage maturation of mammary epithelial cells through regulation of the Erk1/2 pathway. Stem Cell Res. 15, 376–383 (2015).

  38. 38.

    Dimri, M. et al. Modeling breast cancer-associated c-Src and EGFR overexpression in human MECs: c-Src and EGFR cooperatively promote aberrant three-dimensional acinar structure and invasive behavior. Cancer Res 67, 4164–4172 (2007).

  39. 39.

    Harma, V. et al. A comprehensive panel of three-dimensional models for studies of prostate cancer growth, invasion and drug responses. PLoS ONE 5, e10431 (2010).

  40. 40.

    Dent, R. et al. Triple-negative breast cancer: clinical features and patterns of recurrence. Clin. Cancer Res. 13, 4429–4434 (2007).

  41. 41.

    Creighton, C. J. et al. Residual breast cancers after conventional therapy display mesenchymal as well as tumor-initiating features. Proc. Natl Acad. Sci. USA 106, 13820–13825 (2009).

  42. 42.

    Li, X. et al. Intrinsic resistance of tumorigenic breast cancer cells to chemotherapy. J. Natl. Cancer Inst. 100, 672–679 (2008).

  43. 43.

    Goldman, A. et al. Temporally sequenced anticancer drugs overcome adaptive resistance by targeting a vulnerable chemotherapy-induced phenotypic transition. Nat. Commun. 6, 6139 (2015).

  44. 44.

    Gupta, P. B. et al. Identification of selective inhibitors of cancer stem cells by high-throughput screening. Cell 138, 645–659 (2009).

  45. 45.

    Nagathihalli, N. S. & Merchant, N. B. Src-mediated regulation of E-cadherin and EMT in pancreatic cancer. Front Biosci. (Landmark Ed.). 17, 2059–2069 (2012).

  46. 46.

    Fang, D. et al. Epithelial-mesenchymal transition of ovarian cancer cells is sustained by Rac1 through simultaneous activation of MEK1/2 and Src signaling pathways. Oncogene 36, 1546–1558 (2017).

  47. 47.

    Choi, Y. L. et al. LYN is a mediator of epithelial-mesenchymal transition and a target of dasatinib in breast cancer. Cancer Res. 70, 2296–2306 (2010).

  48. 48.

    Ginestier, C. et al. Retinoid signaling regulates breast cancer stem cell differentiation. Cell Cycle 8, 3297–3302 (2009).

  49. 49.

    Herold, C. I. et al. Phase II trial of dasatinib in patients with metastatic breast cancer using real-time pharmacodynamic tissue biomarkers of Src inhibition to escalate dosing. Clin. Cancer Res. 17, 6061–6070 (2011).

  50. 50.

    Finn, R. S. et al. Dasatinib as a single agent in triple-negative breast cancer: results of an open-label phase 2 study. Clin. Cancer Res. 17, 6905–6913 (2011).

  51. 51.

    Fornier, M. N. et al. A phase I study of dasatinib and weekly paclitaxel for metastatic breast cancer. Ann. Oncol. 22, 2575–2581 (2011).

  52. 52.

    Mayer, E. L. & Krop, I. E. Advances in targeting SRC in the treatment of breast cancer and other solid malignancies. Clin. Cancer Res. 16, 3526–3532 (2010).

  53. 53.

    Chen, Y. et al. Combined Src and ER blockade impairs human breast cancer proliferation in vitro and in vivo. Breast Cancer Res. Treat. 128, 69–78 (2011).

  54. 54.

    Zhang, S. et al. Combating trastuzumab resistance by targeting SRC, a common node downstream of multiple resistance pathways. Nat. Med. 17, 461–469 (2011).

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Acknowledgements

The authors thank Dr. Stephen Ethier for help with providing SUM159PT cell line. We would also like to thank Aida Kasei for her help with the animal work.

Author information

Affiliations

  1. Department of Medicine, McGill University Health Center, Cancer Research Program, Montreal, Quebec, H4A 3J1, Canada

    • Jun Tian
    • , Fatmah Al Raffa
    • , Meiou Dai
    • , Alaa Moamer
    • , Ibrahim Y. Hachim
    • , Suhad Ali
    • , Bertrand Jean-Claude
    •  & Jean-Jacques Lebrun
  2. Department of Pathology, McGill University Health Center, Montreal, Quebec, H4A 3J1, Canada

    • Baharak Khadang
    •  & Khldoun Bakdounes
  3. Department of Pathology, St. Mary’s Hospital, Montreal, Quebec, H3T 1M5, Canada

    • Khldoun Bakdounes

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Contributions

J.T. designed, performed experiments, analysed data and drafted the manuscript. F.A.R. was involved in performing PCR and western blot experiments. M.D. contributed to designing and performing experiments. A.M. assisted in 3D cell culture. B.K., I.Y.H. and K.B. were involved in IHC analysis. S.A. and B.J. assisted in designing experiments and editing the manuscript. J.J.L. was involved in research design, data interpretation, supervision of the project and drafting the manuscript.

Competing interests

The authors declare no competing interests.

Ethics approval and consent to participate

The study was performed in accordance with the Declaration of Helsinki.

Funding

This study was funded by grant from the Canadian Institutes for Health Research (CIHR to J.J.L.).

Note

This work is published under the standard license to publish agreement. After 12 months the work will become freely available and the license terms will switch to a Creative Commons Attribution-NonCommercial-Share Alike 4.0 Unported License.

Corresponding author

Correspondence to Jean-Jacques Lebrun.

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DOI

https://doi.org/10.1038/s41416-018-0287-3