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

Figure 4

FHC knockdown improves OVCAR3 response to 6 μM cisplatin by increasing ROS production. (a) Representative plots of Annexin V/7-AAD apoptosis assays in OVCAR3^{Neg Control}, OVCAR3^{Neg Control} (6 μM) cisplatin, OVCAR3^{Neg Control} 10 mM NAC, OVCAR3^{Neg Control} (6 μM) cisplatin/10 mM NAC, OVCAR3^siFHC, and OVCAR3^siFHC (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 OVCAR3^{Neg Control}, OVCAR3^{Neg Control} (6 μM) cisplatin, OVCAR3^{Neg Control} 10 mM NAC OVCAR3^{Neg Control} (6 μM) cisplatin/10 mM NAC, OVCAR3^siFHC, and OVCAR3^siFHC (6 μM) cisplatin, by staining with CellROX® Green Reagent (green). Nuclei were stained with DAPI (blue). (c) Representative western blot of FHC in OVCAR3^{Neg Control}, OVCAR3^{Neg Control} (6 μM) cisplatin, OVCAR3^{Neg Control} 10 mM NAC, OVCAR3^{Neg Control} (6 μM) cisplatin/10 mM NAC, OVCAR3^siFHC, and OVCAR3^siFHC (6 μM) cisplatin. γ-Tubulin was used as internal control. WB analysis was performed three times and results were reproducible.

(a)

(b)

(c)