Fig. 8: ATF3-targeted upregulation of PER2 combined with induction of cuproptosis improves efficacy in OSCC treatment.

A Schematic diagram of subcutaneous injection of SCC25 cells to establish an OSCC model in BALB/c nude mice, and anti-tumor treatment with ATF3 inducer 1 and ES. B Nude mice were injected subcutaneously with SCC25 cells, and after tumor formation, saline, ATF3 inducer 1, ES, and a combination of the two were injected according to Blank-SCC25, SCC25 + ATF3 inducer 1, SCC25 + ES, and SCC25 + ATF3 inducer 1 + ES treatment groups, respectively, and the weight and volume were measured after harvesting tumors on day 28. C Western blotting for PER2 protein expression in Blank-SCC25, SCC25 + ATF3 inducer 1, SCC25 + ES and SCC25 + ATF3 inducer 1 + ES groups. D Copper Colorimetric Assay Kit to measure intra-tumor copper concentrations in the Blank-SCC25, SCC25 + ATF3 inducer 1, SCC25 + ES and SCC25 + ATF3 inducer 1 + ES groups. E Non-denaturing gel electrophoresis assay to detect DLAT oligomers in Blank-SCC25, SCC25 + ATF3 inducer 1, SCC25 + ES and SCC25 + ATF3 inducer 1 + ES groups. F Micro-mitochondrial Complex I and II Activity Assay Kit for detecting activity of electron transport chain complexes I and II in Blank-SCC25, SCC25 + ATF3 inducer 1, SCC25 + ES and SCC25 + ATF3 inducer 1 + ES groups. G IHC assay of Ki67 expression in Blank-SCC25, SCC25 + ATF3 inducer 1, SCC25 + ES and SCC25 + ATF3 inducer 1 + ES groups (n = 5, scale bars = 50 μm). All data represent three replicate independent experiments. Data are presented as mean ± SD. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.