Figure 1
miR-491-5p induces apoptosis in IGROV1-R10 ovarian cancer cells and slow proliferation in SKOV3 ovarian carcinoma cell line. (a) Real-time growth curves monitoring was performed with the xCELLigence System. IGROV1-R10 (left panel) and SKOV3 (right panel) ovarian carcinoma cells were seeded into 96-well E-Plates VIEW, which contain electrodes integrated into the bottom surfaces of each well allowing the measurement of cell index (CI) based on impedance. CI correlates with the area of cells attached to the bottom of the plate. Cells were grown for 24 h before transfection with indicated RNAi (miRNA or siRNA). CI was recorded every 2 h for a time span of 96 h. The combination associating siRNAs targeting BCL-XL (siRNA-BCL-XL) and MCL1 (siRNA-MCL1) was used as a positive control to visualize how cellular cytotoxicity could be visualized using the xCELLigence system. The experiment was performed three times in triplicates. Results are expressed as mean±S.D. on three independent experiments. (b) Cell morphology (upper column of each panel), nuclear morphology after DAPI staining (middle column of each panel) and DNA content histograms obtained by flow cytometry (lower column of each panel) were shown 72 h after transfection of IGROV1-R10 (left panel) and SKOV3 (right panel) ovarian carcinoma cells with miR-491-5p or miR-Ctrl (20 nM). (c) IGROV1-R10 (left panel) and SKOV3 (right panel) cells were transfected with miR-491-5p or miR-Ctrl. Cells were lysed after 72 h and protein extracts were assessed for levels of PARP (native and cleaved forms) and caspases-3 and -9 (pro and cleaved (active) forms) by western blotting. β-actin level is shown as loading control
