H-Ferritin Affects Cisplatin-Induced Cytotoxicity in Ovarian Cancer Cells through the Modulation of ROS

Figure 5

FHC overexpression or NAC treatment reduces OVCAR8 response to cisplatin. (a) Representative plots of Annexin V/7-AAD apoptosis assays in OVCAR8^pc3DNA, OVCAR8^pc3DNA (6 μM) cisplatin, OVCAR8^pc3DNA (6 μM) cisplatin/10 mM NAC, and OVCAR8^pc3FHC (6 μM) cisplatin. Cisplatin treatment was performed for 24 h while NAC treatment was performed for 2 h. FACS plots are representative of single experiments. Values are expressed as of three biological replicates. (b) Immunofluorescence analysis of ROS levels in OVCAR8^pc3DNA, OVCAR8^pc3DNA (6 μM) cisplatin, OVCAR8^pc3DNA (6 μM) cisplatin/10 mM NAC, and OVCAR8^pc3FHC (6 μM) cisplatin, by staining with CellROX® Green Reagent (green). Nuclei were stained with DAPI (blue). (c) Representative western blot of FHC in OVCAR8^pc3DNA, OVCAR8^pc3DNA (6 μM) cisplatin, OVCAR8^pc3DNA (6 μM) cisplatin/10 mM NAC, and OVCAR8^pc3FHC (6 μM) cisplatin. γ-Tubulin was used as internal control. WB analysis was performed three times and results were reproducible.

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