Methods for Monitoring Macroautophagy in Pancreatic Cancer Cells

Methods Mol Biol. 2019:1882:197-206. doi: 10.1007/978-1-4939-8879-2_18.

Abstract

Macroautophagy is a catabolic process through which redundant, aged, or damaged cellular structures are first enclosed within double-membrane vesicles (called autophagosomes), and thereafter degraded within lysosomes. Macroautophagy provides a primary route for the turnover of macromolecules, membranes and organelles, and as such plays a major role in cell homeostasis. As part of the stress response, autophagy is crucial to determine the cell fate in response to extracellular or intracellular injuries. Autophagy is involved in cancerogenesis and in cancer progression. Here we illustrate the essential methods for monitoring autophagy in pancreatic cancer cells.

Keywords: Autophagosome; Autophagy; LC3; Lysosome; Pancreatic cancer; p62.

MeSH terms

  • Animals
  • Autophagosomes / drug effects
  • Autophagosomes / pathology
  • Autophagy*
  • Autophagy-Related Proteins / analysis*
  • Autophagy-Related Proteins / metabolism
  • Carcinogenesis / pathology
  • Cell Culture Techniques / instrumentation
  • Cell Culture Techniques / methods
  • Cell Line, Tumor
  • Chloroquine / pharmacology
  • Disease Progression
  • Electrophoresis, Polyacrylamide Gel / instrumentation
  • Electrophoresis, Polyacrylamide Gel / methods
  • Fluorescent Dyes / chemistry
  • Humans
  • Immunoblotting / instrumentation
  • Immunoblotting / methods*
  • Lysosomes / pathology
  • Macrolides / pharmacology
  • Mice
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / methods
  • Pancreas / cytology
  • Pancreas / pathology
  • Pancreatic Neoplasms / pathology*

Substances

  • Autophagy-Related Proteins
  • Fluorescent Dyes
  • Macrolides
  • Chloroquine
  • bafilomycin A1