Methods for Assessing Apoptosis and Anoikis in Normal Intestine/Colon and Colorectal Cancer

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1765)

Abstract

Caspase-dependent apoptosis, including its distinct cell death subroutine known as anoikis, perform essential roles during organogenesis, as well as in the maintenance and repair of tissues. To this effect, the continuous renewal of the human intestinal/colon epithelium is characterized by the exfoliation by anoikis of differentiated cells, whereas immature/undifferentiated cells may occasionally undergo apoptosis in order to evacuate daughter cells that are damaged or defective. Dysregulated epithelial apoptosis is a significant component of inflammatory bowel diseases. Conversely, the acquisition of a resistance to apoptosis represents one of the hallmarks of cancer initiation and progression, including for colorectal cancer (CRC). Furthermore, the emergence of anoikis resistance constitutes a critical step in cancer progression (including CRC), as well as a limiting one that enables invasion and metastasis.

Considering the implications of apoptosis/anoikis dysregulation in gut physiopathology, it therefore becomes incumbent to understand the functional determinants that underlie such dysregulation—all the while having to monitor, assess, or evidence apoptosis and/or anoikis. In this chapter, methodologies that are typically used to assess caspase-dependent apoptosis and anoikis in intestinal/colonic normal and CRC cells, whether in vivo, ex vivo, or in cellulo, are provided.

Key words

Anoikis Apoptosis Caspase Cell line Cell survival Colon Colorectal cancer DNA laddering Small intestine Regulated cell death 

Notes

Acknowledgments

The author thanks Rona K. Graham (Département de pharmacologie-physiologie, FMSS, U. de Sherbrooke), for access to the LI-Cor Odyssey Fc imaging system and the VICTOR X multilabel plate reader spectrophotometer, and Jean-François Beaulieu (Département d’anatomie et de biologie cellulaire, FMSS, U. de Sherbrooke) for the generous gift of the HIEC-6, Caco-2/15, and HT-29 cell lines. This work was supported in part by a grant from the Natural Sciences and Engineering Research Council (NSERC) of Canada (RGPIN 227935-2013). The author is a Researcher of the Canadian Foundation for Innovation (CFI) and a member of the FRQS-funded Centre de Recherche du CHUS.

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© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Département d’anatomie et de biologie cellulaire, Faculté de médecine et des sciences de la santéUniversité de SherbrookeSherbrookeCanada

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