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

Methods Mol Biol. 2018:1765:99-137. doi: 10.1007/978-1-4939-7765-9_7.

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.

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

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aborted Fetus
  • Anoikis / drug effects
  • Apoptosis / drug effects
  • Apoptosis / genetics
  • Blotting, Western / instrumentation
  • Blotting, Western / methods
  • Caspase 3 / metabolism
  • Cell Culture Techniques / instrumentation
  • Cell Culture Techniques / methods*
  • Cell Line, Tumor
  • Colon / pathology
  • Colorectal Neoplasms / pathology*
  • DNA Fragmentation / drug effects
  • Dimethyl Sulfoxide / pharmacology
  • Electrophoresis, Polyacrylamide Gel / instrumentation
  • Electrophoresis, Polyacrylamide Gel / methods
  • Fluorescent Antibody Technique, Indirect / instrumentation
  • Fluorescent Antibody Technique, Indirect / methods
  • Humans
  • In Situ Nick-End Labeling / instrumentation
  • In Situ Nick-End Labeling / methods*
  • Intestinal Mucosa / pathology
  • Polyhydroxyethyl Methacrylate / pharmacology

Substances

  • Polyhydroxyethyl Methacrylate
  • CASP3 protein, human
  • Caspase 3
  • Dimethyl Sulfoxide