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Assessing Autophagy During Retinoid Treatment of Breast Cancer Cells

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Retinoid and Rexinoid Signaling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2019))

Abstract

Retinoids are derived from vitamin A through a multi-step process. Within a target cell, retinoids regulate gene expression by activating the retinoid acid receptors (RAR) and retinoid x receptors (RXR), which are ligand-dependent transcription factors. Besides its therapeutic use in dermatological disorders, all-trans retinoic acid (ATRA) is successfully utilized to treat acute promyelocytic leukemia (APL) patients. The use of ATRA in APL patients is the first example of clinically useful differentiation therapy. Therapeutic strategies aiming at cancer cell differentiation have great potential for solid tumors, including breast cancer. The few clinical studies conducted with ATRA in breast cancer are rather disappointing. However, these studies did not take into account the heterogeneity of the disease and were conducted on unselected cohorts of patients.

We recently showed that ATRA treatment of breast cancer cells induces autophagy, a highly conserved process aiming at degrading and recycling superfluous or harmful cellular components. In addition, autophagy inhibition significantly increases the therapeutic activity of ATRA. This finding is of fundamental importance, since autophagy has a dual role in cancer. Whereas autophagy may be a protective mechanism during the initial phases of cancer development, it may support cancer cell survival in already established tumors. Furthermore, autophagy can lower or enhance therapeutic efficiency, depending on the tumor type and the anticancer agent considered. Therefore, it is important to investigate the role of autophagy in the context of specific tumors and therapeutic approaches. Accurate autophagy studies are challenging given the dynamic nature of the process and the difficulty of measuring the rate of autophagosome degradation (autophagic flux). In this chapter, we provide protocols for a careful assessment of the autophagic flux in ATRA treated 2D and 3D breast cancer cultures.

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Acknowledgments

Deborah Shan-Krauer is gratefully acknowledged for excellent technical support. This research project was supported by the University of Bern Initiator Grant and grants from the Bernese Cancer League, the Werner and the Hedy Berger-Janser Foundation for Cancer Research (to A.M.S.) and Swiss Cancer Research (KFS-3409-02-2014 to M.P.T.).

Conflict of Interest: Manuele G. Muraro is a shareholder and scientific advisor of Cellec Biotek AG. Giulio C. Spagnoli is a Cellec Biotek AG shareholder.

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Authors and Affiliations

  1. Division of Experimental Pathology, Institute of Pathology, University of Bern, Bern, Switzerland

    Sarah Parejo, Mario P. Tschan & Anna M. Schläfli

  2. Graduate School for Cellular and Biomedical Sciences, University of Bern, Bern, Switzerland

    Mario P. Tschan

  3. Department of Biomedicine, University of Basel and University Hospital Basel, Basel, Switzerland

    Manuele G. Muraro & Giulio C. Spagnoli

  4. Laboratory of Molecular Biology, Istituto di Ricerche Farmacologiche Mario Negri IRCCS, Milan, Italy

    Enrico Garattini

  5. National Research Council, Institute of Translational Pharmacology, Rome, Italy

    Giulio C. Spagnoli

Authors
  1. Sarah Parejo
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  2. Mario P. Tschan
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  3. Manuele G. Muraro
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  4. Enrico Garattini
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  5. Giulio C. Spagnoli
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  6. Anna M. Schläfli
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Corresponding author

Correspondence to Anna M. Schläfli .

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Editors and Affiliations

  1. Department of Pathology, Microbiology, and Immunology, University of South Carolina School of Medicine, Columbia, SC, USA

    Swapan K. Ray

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Parejo, S., Tschan, M.P., Muraro, M.G., Garattini, E., Spagnoli, G.C., Schläfli, A.M. (2019). Assessing Autophagy During Retinoid Treatment of Breast Cancer Cells. In: Ray, S. (eds) Retinoid and Rexinoid Signaling . Methods in Molecular Biology, vol 2019. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9585-1_17

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  • DOI: https://doi.org/10.1007/978-1-4939-9585-1_17

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9584-4

  • Online ISBN: 978-1-4939-9585-1

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