Fig. 5: c-Myc/ORC2 axis is the crucial signaling pathway underlying the response to anlotinib and KRAS-G12Ci combined treatment.

A Heatmap of the mRNA expression of SNRPD2, RPL22, DEK, EIF4E, COPS5, ORC2, SSB, UBE2E1, AP3S1 in five cell lines (Calu-1, SW1573, H23, H2122SR, and H2030SR) detected by qPCR. n = 3 per group. B The expression of eIF4E, COPS5, and ORC2 in H2122SR and H2030SR compared with parental cell lines detected by western blotting. C Survival plot of 154 KRAS-mutant NSCLC patients with different mRNA expression status of MYC and ORC2. Statistical differences are assessed using the log-rank test. Data are from TCGA Pan-Cancer Atlas. D The expression of ORC2 in five cell lines (Calu-1, SW1573, H23, H2122SR, and H2030SR) treated with anlotinib (2 μM) plus sotorasib (1 μM) for 24 h detected by western blotting. Sot sotorasib, Anlo anlotinib. E Cell viability assays of H2122 and H2030 after MYC overexpression and ORC2 knockdown (si-RNA 20 μM, 72 h). Results are shown as mean ± SEM. F–I EdU/PI staining assays of H23 (F, G) and H2122SR (H, I) treated with anlotinib (2 μM) plus sotorasib (1 μM) for 24 h after transfected with EV, MYC-OE, si-ORC2 #1 or si-ORC2 #2 (20 μM, 72 h). Results are shown as mean ± SEM. Statistical differences are determined using one-way ANOVA with Tukey’s multiple comparisons test. Bold: p < 0.05. n = 3 per group. Combo: anlotinib plus sotorasib. J–M Cell growth and cell viability assays of H23 (J, K) and H2122SR (L, M) treated with anlotinib (2 μM) plus sotorasib (1 μM) for 24 h EV, MYC-OE, si-ORC2 #1 or si-ORC2 #2 (20 μM, 72 h). Results are shown as mean ± SEM. Statistical differences are determined using one-way ANOVA with Tukey’s multiple comparisons test. Bold: p < 0.05. n = 3 per group. Combo: anlotinib plus sotorasib.