Amplicon-Dependent CCNE1 Expression Is Critical for Clonogenic Survival after Cisplatin Treatment and Is Correlated with 20q11 Gain in Ovarian Cancer
Figure 3
Combined siRNA knock-down and cisplatin treatment in ovarian tumor cell lines.
(A) Cell viability after transfection with individual siRNAs and cisplatin treatment normalized to cisplatin-treated control cells without siRNA. Cisplatin dose of 3 µM or 6 µM was used for OVCAR-3 and SK-OV-3 cells respectively. Average normalized absorbance (490 nm) from three independent MTS assays (triplicate wells per condition) and SEM plotted (n = 3). (B) Cisplatin dose response after transfection with CCNE1 or non-silencing siRNA in OVCAR-3 and SK-OV-3 cell lines. Arrow indicates drug treatment dose used in initial screen; p-value indicates significance of difference between fitted curves. Average normalized MTS assay absorbance to cells without cisplatin treatment, SEM and four-parameter fitted Hill slope plotted (n = 3 for each drug concentration). (D) Cell viability after transfection with CCNE1 or NS siRNAs and cisplatin treatment normalized to cisplatin-treated no siRNA control cells for each cell line. Statistical significance calculated by comparison to NS siRNA, cisplatin-treated cells in the same cell line. Average normalized absorbance from MTS assay and SEM plotted (n = 3). See Table S4 for cisplatin treatment doses.