Biochemical and Biophysical Research Communications, Vol.427, No.1, 11-17, 2012
Use of inducible Atg5 deletion and expression cell lines in study of the pro-survival function of autophagy under starvation
At present the role of autophagy in cell death and cell survival remains controversial, partly owning to the contradictory results from the immortalized mouse embryonic fibroblasts (MEFs) with knockout of different autophagy-related genes (Atg). Here we aimed to reexamine the role of autophagy in cell death under starvation and other stress conditions. First, different clones of Atg5 knockout MEFs had different susceptibility to stress-mediated cell death, indicating that it is the clonal variation, rather than the deficiency of Atg5 or autophagy per se that determines the susceptibility. Next, we tested two cell lines with inducible Atg5 deletion or expression and demonstrated that cells without Atg5 expression were more sensitive to starvation-induced apoptosis. Finally, we found that chloroquine was only effective in sensitizing starvation-induced cell death in Atg5-expressing cells, but not in Atg5-deficient cells. Such observations thus provide unequivocal evidence supporting the pro-survival function of autophagy under starvation. Moreover, our data demonstrate the usefulness of cells with inducible deletion or expression of Atg in the study of autophagy in cell death and cell survival. (c) 2012 Elsevier Inc. All rights reserved.