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Measuring Autophagy in the Context of Cancer

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Tumor Microenvironment

Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 899))

Abstract

Autophagy plays multiple roles in the formation and progression of cancer, including both suppressive and promotive roles. It not only impacts cancer cell growth and viability directly but also has a significant role through its effects on the tumor microenvironment. Measurement of autophagy can be confusing and sometimes misleading due to the inherent difficulty of measuring both the formation and turnover of molecules involved in the autophagic process. The LC3 proteins serve as autophagosomal markers and are the basis for most of the assays used for measuring autophagy. Since each of the current assays for autophagy has significant limitations, the use of multiple assays for the analysis of autophagy in most contexts is highly advised. Here we outline three assays that are commonly used to evaluate autophagic flux in cells. These assays include the determination of LC3II formation and LC3II and p62 turnover by use of Western Blotting, quantification of LC3 puncta, and the measurement of autophagic flux using tandem labeled mCherry-GFP-LC3.

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Author information

Authors and Affiliations

  1. Department of Pharmacology, University of Colorado School of Medicine, 12801 East 17th Avenue, Rm. L18-6110, MS 8303, Aurora, CO, 80045, USA

    Michael J. Morgan Ph.D.

  2. Department of Pharmacology, University of Colorado School of Medicine, 12801 East 17th Avenue, Rm. L18-6105, MS 8303, Aurora, CO, 80045, USA

    Andrew Thorburn

Authors
  1. Michael J. Morgan Ph.D.
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  2. Andrew Thorburn
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Corresponding author

Correspondence to Michael J. Morgan Ph.D. .

Editor information

Editors and Affiliations

  1. Perelman School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA

    Constantinos Koumenis

  2. Cell & Developmental Biology, Oregon Health & Science University, Portland, Oregon, USA

    Lisa M. Coussens

  3. Department of Radiation Oncology, Stanford University School of Medic, Stanford, California, USA

    Amato Giaccia

  4. Department of Oncology, University of Oxford, Oxford, Oxfordshire, United Kingdom

    Ester Hammond

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© 2016 Springer International Publishing Switzerland

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Morgan, M.J., Thorburn, A. (2016). Measuring Autophagy in the Context of Cancer. In: Koumenis, C., Coussens, L., Giaccia, A., Hammond, E. (eds) Tumor Microenvironment. Advances in Experimental Medicine and Biology, vol 899. Springer, Cham. https://doi.org/10.1007/978-3-319-26666-4_8

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