Methods in Enzymology 2009-01-01

Detachment-induced autophagy in three-dimensional epithelial cell cultures.

Jayanta Debnath

Index: Meth. Enzymol. 452 , 423-39, (2009)

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Abstract

Integrin-mediated cell adhesion to extracellular matrix (ECM) is critical for normal epithelial cell survival; cells deprived of ECM contact rapidly undergo apoptotic cell death, termed anoikis. Recent work demonstrates that ECM detachment also robustly induces autophagy, which protects epithelial cells from the stresses of matrix detachment, allowing them to survive provided they can reattach in a timely manner. This chapter details the methods used to measure and manipulate detachment-induced autophagy during lumen formation in three-dimensional in vitro glandular epithelial cultures as well as in traditional substratum detachment assays employed to study anoikis.