Fig. 2.
In vitro and in vivo synthetic lethality in ARID1A-KO HCT116 cells by AURKA inhibition. a Synthetic lethality in ARID1A−/− HCT116 cells by AURKAi. ARID1A+/+ HCT116 or three ARID1A−/− clones were treated with 1 μM AURKAi for 72 h and nuclei were stained with Hoechst 33342. Scale bars, 100 µm. b Nuclear density was measured with Image J software as a surrogate for cell viability. Error bars represent s.d. **P < 0.01, Student’s t test. c Silencing of AURKA expression in ARID1A isogenic cell pair by siRNA (siAURKA#1). GAPDH was used as a loading control. d Synthetic lethality in ARID1A−/− HCT116 cells by AURKA siRNA. ARID1A+/+ or ARID1A−/− clone was transfected with various concentrations of AURKA siRNA for 72 h and the cell images were taken with IncuCyte ZOOM. Scale bars, 300 µm. e Integrated density was measured with the IncuCyte ZOOM software as a surrogate for cell viability. Error bars represent s.d. **P < 0.01, Student’s t test. f Ectopic overexpression of ARID1A (pLenti-ARID1A) in ARID1A−/− HCT116 cells. g Overexpression of ARID1A reversed the synthetic lethality effect by AURKAi. ARID1A+/+ HCT116, ARID1A−/− HCT116, and ARID1A−/− HCT116 transfected with an ARID1A plasmid were treated with AURKAi for 72 h, and the cell viability was assessed with AlamarBlue assay. Error bars represent s.d. **P < 0.01, Student’s t test. h Schematic illustration of mouse tumor xenograft experiments with HCT116 ARID1A isogenic cell pair. i, j Tumor growth curve in nude mice bearing ARID1A+/+ HCT116 (i) or ARID1A−/− HCT116 (j) xenografts after injection of vehicle or 60 mg kg−1 (mpk) AURKAi. Error bars represent s.d. *P < 0.05 between vehicle and AURKAi treatment groups (n = 5), Student’s t test. k, l Wet weight measurement of the tumors isolated from mice bearing ARID1A+/+ HCT116 (k) or ARID1A−/− HCT116 (i) xenografts at 24 days after injection of vehicle, 30 or 60 mpk AURKAi. Error bars represent s.d. *P < 0.05 between vehicle and 60 mpk AURKAi treatment groups (n = 5), Student’s t test