Intracellular
iron content was investigated by using the fluorescent probe Phen
Green SK (PGSK; Invitrogen) and
FerroOrange (Dojindo Laboratories)
according to the manufacturer’s instructions.. SKOV3 and OVCAR3
cancer cells were treated (control, olaparib, or olaparib-Ga) for
3 h, then incubated with 5 mM PGSK or 0.5 μg/mL
FerroOrangeat 37 °C for 30 min. For PGSK, the cells were then washed with
PBS twice (washing was not necessary after
FerroOrange treatment).
Fluorescence images were recorded
via a confocal
laser scanning microscope (Olympus,
Fluoview FV1200). The experiments
were repeated independently three times.
Intracellular ROS generation
was measured using a confocal laser scanning microscope (CLSM). SKOV3
and OVCAR3 cancer cells were cultured in CLSM culture dishes (2 ×
10
5 cells per well) overnight under normal culture conditions.
The culture medium was then replaced with an equal volume of DMEM
containing control, olaparib, or olaparib-Ga and incubated for 3 h.
PBS containing 2.5 μM ROS was used to replace the medium, and
the cells were then incubated at 37 °C for 30 min in the dark.
The cells were then washed three times. Green fluorescence was detected
to confirm intracellular ROS generation using a confocal laser scanning
microscope (Olympus,
Fluoview FV1200). The experiments were performed
independently three times.
Li Y., Cen Y., Fang Y., Tang S., Li S., Ren Y., Zhang H., Lu W, & Xu J. (2022). Breaking the Iron Homeostasis: A “Trojan Horse” Self-Assembled Nanodrug Sensitizes Homologous Recombination Proficient Ovarian Cancer Cells to PARP Inhibition. ACS Nano, 16(8), 12786-12800.
