Fig. 2
Anti-tumor effects of FAST and other treatments in cancer cells and the WEHI-3 xenograft mice. Cells (HepG2, HL7702, KG-1a) were transfected by pDMP-NT/T2/T5/T7 overnight respectively, and then incubated with or without 50 µg/mL FeNPs for 48 h. a Cellular iron content post treatment. b Total ROS levels post treatment. a and b use a same set of symbols. All data are presented as mean ± s.d (n = 3 wells). pDMP-NT, a plasmid coding an artificial microRNA targeting no transcripts (NT); pDMP-T2, a plasmid coding artificial microRNAs targeting two target (T) genes including FPN and LCN2; pDMP-T5, a plasmid coding artificial microRNAs targeting five target genes including FSP1, FTH1, GPX4, SLC7A11, and NRF2; pDMP-T7, a plasmid coding artificial microRNAs targeting seven target genes including FSP1, FTH1, GPX4, SLC7A11, NRF2, FPN, and LCN2. c Quantitative clone formation assays. Cells (PANC-1, MDA-MB-453, B16F10, CT26) were infected with rAAV-NT/T5/T7 at the dose of 1 × 105 vg per cell for 24 h and then incubated with 50 µg/mL FeNPs for another 48 h. PBS, cells just treated with phosphate buffered saline (PBS) as a control. After treatment, 200 cells were seeded into 6-well plate and cultured until colonies were clearly visible (> 50 cells) (about two weeks). Each treatment was conducted in triplicates. The images of cell clones are shown in Additional file 1: Fig. S20. rAAV-NT/T5/T7, rAAVs packaged with pDMP-NT, pDMP-T5, and pDMP-T7. d Schematics of animal treatment (WEHI-3 xenograft mice). s.c., subcutaneous injection; i.v., intravenous injection. e Tumor imaging. f Tumor growth curve. All data are presented as mean ± s.d (n = 7 mice). g Tumor weight. (n = 7 mice). h Kaplan-Meier survival curve (n = 10 mice). The statistical significance was analyzed by the log-rank test. i Expression of stemness- and proliferation-related marker genes. The relative expression detected by qPCR (2−ΔΔt) was shown by heatmap. The p values between rAAV-T5 + FeNPs and rAAV-T7 + FeNPs are given. j Representative H&E-stained section of tumors. k Iron content in tumors of mice in different treatment groups (n = 7 mice). By these comparative treatments of cells and mice, the treatment of pDMP/rAAV-T7 + FeNPs that shows the best anti-tumor effect was defined as FAST, referring to Ferroptosis ASsassinates Tumor