Modulating lysosomal function through lysosome membrane permeabilization or autophagy suppression restores sensitivity to cisplatin in refractory non-small-cell lung cancer cells
Fig 8
Inhibition of autophagy using ATG5 siRNA potentiates the activity of cisplatin against A549cisR cells.
(A) A549cisR cells were treated for 24 hours with 25 μM cisplatin prior to transfection with ATG5 siRNA or NT siRNA control. Following two days incubation, media was removed and cells were treated with different cisplatin concentrations for an additional 24 or 48 hours. At the end of the incubation, cell viability was measured using CellTiter-Blue® reagent. Experiments were performed in 1%FBS media. The X-axis represents the percentage of cell viability at 24 hours (T24/T1) normalized to the negative control 1%FBS. Error bars represent SEM from three independent experiment. p-value corresponding to the difference between ATG5 and NT siRNA-transfected cells was obtained by 2-way ANOVA. p-value corresponding to the individual difference between ATG5 and NT siRNA-transfected cells treated with the same concentration of cisplatin was calculated by the Sidak’s post-test analysis, with * annotating a p-value <0.05. (B, C) A549cisR cell lysates were prepared either immediately at the end of the siRNAs (ATG5 or NT) transfections or two days following the end of the transfection (annotated as 2d). Western Blot analysis was performed for the detection of ATG5, LC3, p62 and α-tubulin proteins.