9mm tumors(lower panel; shown as means±SEM,n values indicate the number of mice used for each condition and are labeled in each panel; P values were calculated by two-way ANOVA followed by Tukey's test).c Aldometanib effectively inhibits late-stage HCC. Mice were induced to develop HCC using DEN and HFD as in a. Aldometanib at 100 mg/L was administered in drinking water starting at 12 weeks, 24 weeks, or 35 weeks of age(depicted in the upper panel). Numbers of tumors in each size/diameter category for each entry time of aldometanib are shown(lower panel; shown as means±SEM, n values represent the number of mice for each starting age and are labeled in each panel, with P values calculated by two-way ANOVA followed by Tukey's test).d Aldometanib suppresses AFP levels in DEN-HFD mice. HCC tissues from aldometanib-treated and untreated DEN-HFD mice, prepared as in b,were subjected to immunohistochemistry(IHC) staining for AFP. Representative images at different magnifications are shown in the upper panel, and the percentages of AFP-positive area in the HCC tissues were calculated and are shown in the lower panel(means±SEM, n= 4(vehicle) or 5(aldometanib) mice for each treatment, with P values calculated by two-sided Student's t-test).e Aldometanib inhibits HCC in MYC;Trp53-/- mice. Wild-type BALB/c mice(12 weeks old) were hydrodynamically injected via the tail vein with plasmids carrying sgRNAs targeting Trp53 for knockout, along with a plasmid for Myc overexpression. One week later, mice were intraperitoneally injected with CCl4 twice a week for 3 weeks(depicted in the upper panel). Aldometanib was administered at 100 mg/L in drinking water starting at 16 weeks of age. At week 18, mice were euthanized, and the numbers of tumors in each size/diameter category are shown(lower panel; shown as means±SEM, n numbers are labeled in each panel, with P values calculated by two-way ANOVA followed by Sidak's test). f Aldometanib reduces the size of orthotopic allografts derived from Hepa1-6 hepatoma cells. Hepa1-6 cells were transplanted into the left liver lobe of C57BL/6 mice to form orthotopic allografts. At day 2 post-transplantation, the mice were treated with aldometanib in drinking water for 15 days(depicted in the upper panel). On day 17, mice were euthanized, and the weights of the orthotopic allografts were determined(lower panel; shown as means±SEM, n values represent the number of mice and are labeled in each panel, with P values calculated by two-sided Student's t-test). g Aldometanib treatment increases the median lifespan of mice bearing Hepa1-6 allografts. Wild-type C57BL/6 mice were transplanted with Hepa1-6 cells as in f, and then treated with aldometanib in drinking water on day 2 post-transplantation(depicted in the upper panel). The lifespans of the mice were determined and are shown as Kaplan-Meier curves(see also statistical analyses in Supplementary information, Table S2). Experiments in this figure were performed three times.'>非癌性宿主肝中AMPK在aldometanib抑制HCC中起主导作用通过肝脏特异性AMPKα敲除（AMPKα-LKO）小鼠实验，研究人员发现aldometanib的抗肿瘤效果完全依赖于AMPK的激活。特别有趣的是，当将AMPKα-/- Hepa1-6细胞移植到野生型小鼠肝脏时，aldometanib仍能有效抑制肿瘤生长；反之，如果将野生型Hepa1-6细胞移植到AMPKα-LKO小鼠肝脏，aldometanib的抑瘤效果显著减弱。这表明非癌性宿主肝组织中的AMPK在aldometanib介导的肿瘤抑制中起主导作用。
