A Ploidy Increase Promotes Sensitivity of Glioma Stem Cells to Aurora Kinases Inhibition
Danusertib induces senescence/autophagy in sensitive cell line. (a) GBM2 (sensitive) and G166 (resistant) cell lines were treated with 500 nM Danusertib for 48 h. After the incubation, they were fixed and stained for β-galactosidase to evaluate induction of senescence. Representative images showing β-galactosidase staining at baseline and after Danusertib treatment are reported. The graph shows the percentages of senescent cells in the two cell lines. Danusertib induced a significant and more relevant increase in the percentage of β-gal positive cells only in the sensitive cell line (GBM2). An average of 100 cells/condition/experiment were randomly imaged and scored. Results are representative of three independent experiments. Error bars represent SEM. t-test was performed on raw data: p<0,05; p<0,01. (b) Beclin level and LC3-I and LC3-II ratio were determined in untreated and 48 h 500 nM treated GBM2 (sensitive) and G166 (resistant) cell lines in order to evaluate the activation of an autophagic process. Representative images of Western blot analysis and quantitative data are reported. Beclin, LCR-I and II levels were determined and normalized on GAPDH. All the values are expressed in Arbitrary Unit (AU). Danusertib induced a non-statistically significant increase of Beclin level and LC3-I and LC3-II ratio only in the sensitive cell line (GBM2). Results are representative of three independent experiments. Error bars represent SEM.
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