Identification of a Potential Ovarian Cancer Stem Cell Gene Expression Profile from Advanced Stage Papillary Serous Ovarian Cancer
Figure 5
Validation of SP cells from cell lines and the effect of GSI-IX inhibitor on SP cells colony forming efficiency.
(A) Repopulation Assay: The SKOV3 and A224 cells were stained with Hoechst 33342 dye and sorted for SP and MP populations. The cells were cultured for 8 days for repopulation, and then reanalyzed by flow cytometry. This demonstrated the enrichment of SP cells (22.5% and 10.4% for SKOV3 and A224 resp.) with a capacity to regenerate to MP cells. (B, C, D) qRT-PCR analysis for stem cell marker genes and transporter genes (p<0.01). To calculate the relative expression for each gene, the 2−ΔΔCT method was used averaging the CT values for the three housekeeping genes (Cyclophilin A, GUSB, GAPDH) for a single reference gene value. (E, F) Inhibitory effect of GSI-IX on Colony forming efficiency of sorted SP and MP cells from SKOV3. Cells were sorted to SP and MP populations and treated with 10 and 20 µg of GSI-IX. The inhibitor carrier DMSO is used as a control.