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Detection of Anoikis Using Cell Viability Dye and Quantitation of Caspase Activity

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Detection of Cell Death Mechanisms

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2255))

Abstract

Anoikis is a type of programmed cell death triggered by the loss of cellular interaction with the extracellular matrix (ECM) and culminates in the activation of caspases. Specific interaction between cellular receptors such as integrins and the ECM is important to maintain cellular homeostasis in normal tissues through multiple cascades. This interaction provides not only physical attachment, but more importantly, vital interaction with the actin cytoskeleton and growth factors. Normal epithelial and endothelial cells require this interaction with ECM to survive. In cancer, the acquisition of anoikis resistance is a hallmark of malignant transformation and is required in the process of metastasis formation. As such, strategies to inhibit and/or counteract anoikis resistance are important in controlling cancer progression. In this chapter, we describe the method for detecting anoikis using cell viability and caspase activity assays.

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References

  1. Elmore S (2007) Apoptosis: a review of programmed cell death. Toxicol Pathol 35(4):495–516. https://doi.org/10.1080/01926230701320337

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. McIlwain DR, Berger T, Mak TW (2013) Caspase functions in cell death and disease. Cold Spring Harb Perspect Biol 5:a008656

    Article  Google Scholar 

  3. Paoli P, Giannoni E, Chiarugi P (2013) Anoikis molecular pathways and its role in cancer progression. Biochim Biophys Acta 1833(12):3481–3498. https://doi.org/10.1016/j.bbamcr.2013.06.026

    Article  CAS  PubMed  Google Scholar 

  4. Kim Y-N, Koo KH, Sung JY, Yun U-J, Kim H (2012) Anoikis resistance: an essential prerequisite for tumor metastasis. Int J Cell Biol 2012:306879. https://doi.org/10.1155/2012/306879

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Gilmore AP (2005) Anoikis. Cell Death Differ 12(2):1473–1477. https://doi.org/10.1038/sj.cdd.4401723

    Article  CAS  PubMed  Google Scholar 

  6. Reddig PJ, Juliano RL (2005) Clinging to life: cell to matrix adhesion and cell survival. Cancer Metastasis Rev 24(3):425–439. https://doi.org/10.1007/s10555-005-5134-3

    Article  PubMed  Google Scholar 

  7. Nagaprashantha LD, Vatsyayan R, Lelsani PCR, Awasthi S, Singhal SS (2011) The sensors and regulators of cell-matrix surveillance in anoikis resistance of tumors. Int J Cancer 128(4):743–752. https://doi.org/10.1002/ijc.25725

    Article  CAS  PubMed  Google Scholar 

  8. Wang P, Valentijn AJ, Gilmore AP, Streuli CH (2003) Early events in the Anoikis program occur in the absence of caspase activation. J Biol Chem 278(22):19918–19925

    Google Scholar 

  9. Frisch SM, Francis H (1994) Disruption of epithelial cell-matrix interactions induces apoptosis. J Cell Biol 124(4):619–626. https://doi.org/10.1083/jcb.124.4.619

    Article  CAS  PubMed  Google Scholar 

  10. Simpson CD, Anyiwe K, Schimmer AD (2008) Anoikis resistance and tumor metastasis. Cancer Lett 272(2):177–185. https://doi.org/10.1016/j.canlet.2008.05.029

    Article  CAS  PubMed  Google Scholar 

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Correspondence to Roslyn Tedja .

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Tedja, R., Fox, A., Alvero, A.B. (2021). Detection of Anoikis Using Cell Viability Dye and Quantitation of Caspase Activity. In: Alvero, A.B., Mor, G.G. (eds) Detection of Cell Death Mechanisms. Methods in Molecular Biology, vol 2255. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1162-3_7

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  • DOI: https://doi.org/10.1007/978-1-0716-1162-3_7

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1161-6

  • Online ISBN: 978-1-0716-1162-3

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