Fig. 9
Fluphenazine-mediated inhibition of HCC growth is dependent on iron and KLF14. A Cell apoptosis induced by fluphenazine (10 μM) were decreased with FAC (100 μM) treatment. B Cell growth curve of HepG2 and Huh7 cells were measured in media supplemented with PBS, fluphenazine (10 μM), or their combination with FAC for 4 days. C Western blotting tested the expression of KLF14, IRP2, TfR1 and FH in KLF14-silenced cells with fluphenazine (10 μM) treatment. D KLF14-silenced cells were incubated with 10 μM fluphenazine for 36 h. The relative cellular LIP content were measured using calcein-AM assay. E Silencing KLF14 reduced the cell apoptosis induced by fluphenazine (10 μM). F Cell growth curve of KLF14-silenced HepG2 and Huh7 cells were measured in media supplemented with PBS or fluphenazine (10 μM) for 4 days. G HepG2 cells were grafted into nude mice subcutaneously. When the tumor volume reached approximately 100 mm3, intraperitoneal injection of fluphenazine (8 mg/kg) was followed before harvesting solid tumors. Xenograft tumors were analyzed at indicated time points. n = 6. H Tumor volume was measured starting 7 days after inoculation. I, J Tumors were isolated and weight was measured. n = 6. K Immunohistochemistry staining for H&E, Perl’s Blue (iron), Ki67, cleaved caspase3, KLF14 and IRP2 in tumor tissues isolated from mice (scale bar 50 μm). All the results are presented as means ± SD from three independent experiments. Two-tailed unpaired Student’s T-tests were performed. n.s, not significant, *P < 0.05, **P < 0.01 and ***P < 0.001