Figure 3
Pitavastatin induces autophagy associated cell death for GBM. (A) Pitavastatin induced SK72 glioma primary cell death in adherent and neurosphere cultures. Note rounding of cells at 24 h. (B) FACS analysis showing that pitavastatin treated U87 cells were arrested in G0/G1 phase with a much lower percentage of the total population in S phase. (C) In GFP-LC3 transduced-U87 GBM cells, pitavastatin induced a punctuate GFP-LC3 distribution, which is a characteristic of autophagy. (D) Western blot showing that pitavastatin induced the transformation of LC3 protein from isoform I to II in U87, U251 and SK72 cells. (Pitavastatin concentrations in μM). LC3I/II was calculated with NIH ImageJ software (E) Caspase-3/7 activity in U87 cells induced by staurosporine for 8 hours detected by the Apo-One fluorescence kit. In contrast, no caspase 3/7 activity was seen with pitavastatin treatment during 2 days. Inset: Western blot illustrating that there was no detectable cleavage (CL) of caspase-3 after pitavastatin treatment (P, 10 μM); Staurosporine treatment (S, 0.5 μM) was used as a positive control and was associated with caspase-3 cleavage (CL).