Assessing Autophagy During Retinoid Treatment of Breast Cancer Cells.

Parejo, Sarah; Tschan, Mario; Muraro, Manuele G; Garattini, Enrico; Spagnoli, Giulio C; Schläfli, Anna M (2019). Assessing Autophagy During Retinoid Treatment of Breast Cancer Cells. In: Ray, Swapan K. (ed.) Retinoid and Rexinoid Signaling. Methods in molecular biology: Vol. 2019 (pp. 237-256). Springer 10.1007/978-1-4939-9585-1_17

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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.

Item Type:

Book Section (Book Chapter)


04 Faculty of Medicine > Service Sector > Institute of Pathology > Tumour Pathology

Graduate School:

Graduate School for Cellular and Biomedical Sciences (GCB)

UniBE Contributor:

Parejo, Sarah and Tschan, Mario


500 Science > 570 Life sciences; biology






Methods in molecular biology






Mario Tschan

Date Deposited:

27 Aug 2019 12:16

Last Modified:

27 Aug 2019 12:16

Publisher DOI:


PubMed ID:


Uncontrolled Keywords:

ATRA Autophagic flux Autophagy Bioreactor Breast cancer Immunofluorescence Retinoids Western blot mCherry-eGFP-LC3B


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