Ca 2+ -dependent autophagy is enhanced by the pharmacological agent PK11195.
The 1-(2-Chlorophenyl-N-methylpropyl)-3-isoquinolinecarboxamide, PK11195, is a proven enhancer of apoptotic cell death in a variety of cellular models. Recently, we have shown that by targeting the oncogene Bcl-2, PK11195 increases the [Ca ( 2+) ] in the Endoplasmic Reticulum ([Ca ( 2+) ]er) as well as IP3 induced mitochondrial ([Ca ( 2+) ]m) and cytosolic ([Ca ( 2+) ]c) Ca ( 2+)  transients in HeLa cervix carcinoma cells. Here, in the same cells, we have investigated PK11195 contribution to models of pharmacologically induced macroautophagy. To do so, we have monitored the pattern of LC3 (the mammalian orthologue of yeast Atg8) distribution and post transcriptional modifications after challenging with Ca ( 2+) -dependent (ATP, Vitamin D3) and independent (Rapamycin and H 2O 2) stimuli for autophagy execution. We found that PK11195 plays a pro-autophagy role if associated with ATP and Vitamin D3 to be ineffective if co-incubated with Rapamycin and H 2O 2. Notably, Bcl-2 deletion abolished PK11195 effects thus suggesting a selective way of action against the oncogene. By these means, PK11195 is proposed as facilitator of Ca ( 2+)  mediated autophagy and tool to ascertain the Bcl-2 contribution to the onset and unfolding of this essential catabolic process for cellular homeostasis.
Gastaldello
Annalisa
A
Royal Veterinary College, University of London, London UK.
Callaghan
Holly
H
Gami
Priya
P
Campanella
Michelangelo
M
D001343:Autophagy / Q000187:drug effects*; D002118:Calcium / Q000494:pharmacology*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D007546:Isoquinolines / Q000494:pharmacology*; D010588:Phagosomes / Q000187:drug effects / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism
D007546:Isoquinolines; D019253:Proto-Oncogene Proteins c-bcl-2; D049452:Green Fluorescent Proteins; D002118:Calcium; C037850:PK 11195