Fig. 4.

Targeting E2F1 and SMAD3 could extensively down-regulate subtype-specific signature genes and exert selective vulnerability on different subtypes of liver cancer. (A) The expressions of subtype-specific signature genes and the featured molecules belonging to several developmental signaling pathways were detected at different liver developmental stages. (B) E2F1- and (C) SMAD3-specific small-molecule inhibitors were used to treat HCC cell lines. The relative expression of their subtype-specific signature genes and developmental signaling pathways were detected by qPCR. The heat map represents three independent experiments. *P < 0.05. (D) HCC cells with ES-like signature (ES+/LP−), LP-like signature (ES−/LP+), or mixed signature (ES+/LP+) were treated with E2F1 and SMAD3 inhibitors at indicated concentrations. The percentage of cell growth inhibition was detected. *P < 0.05, **P < 0.01 by independent t test. (E) Tumor organoids derived from primary HCC patients with different oncofetal properties were treated with E2F1 or SMAD3 inhibitors, and their sensitivities to drug treatment were examined. *P < 0.05, ***P < 0.001, independent t test. ns, not significant. (F) The correlations of ES index (z-score) and LP index (z-score) of the primary HCC organoids with percentage of drug inhibitory rates.