Fig. 5: FBXO32 promotes tumor growth and metastasis in vitro and in vivo.

a–f Huh7 or CFPAC-1 cells (5 × 10⁶) stably expressing sh-FBXO32 or sh-NC were injected subcutaneously to axilla of BALB/c nude mice. After 4 weeks, the masses of tumor formed from Huh7 or CFPAC-1 cells were divided equally into 1 mm³ pieces and implanted orthotopically into BALB/c nude mice (n = 6). The mice were sacrificed 4 weeks later, then the tumor volumes were monitored (a, d). H&E staining was performed to determine the tumor tissue, and immunohistochemistry (IHC) staining was utilized to measure the protein levels of FBXO32 and Ki-67 (b, e). Scale bar, 50 μm. Representative lung (or liver) metastasis nodule images were presented, and the incidence of lung (or liver) metastasis from liver (or pancreatic) cancer was calculated. H&E and IHC staining were used to determine the metastatic tumor from liver or pancreas (c, f). Top scale bar, 200 μm, middle and bottom scale bar, 50 μm. g, h Hepa1-6 cells with constitutively expressing luciferase were injected orthotopically into C57BL/6N mice (n = 4). Then AAV-shFbxo32 and AAV-shNC were injected intraperitoneally into the mice after one week. Bioluminescence device was utilized to monitor the tumor size at the first and fifth week, and representative bioluminescence images were shown (g). QPCR was performed to detect Fbxo32 mRNA level of tumor tissues from three groups of mice (h). i–k CCK-8 and colony formation assays were used to measure the proliferation of Huh7 and CFPAC-1 cells after infected with LV-shFBXO32 and LV-shNC, or transfected with OE-FBXO32 and control plasmids. (n = 3 independent experiments). l, m Migration (l) and invasion (m) assays were performed to determine the metastasis of Huh7 and CFPAC-1 cells with stable expression of sh-FBXO32 or sh-NC by lentivirus infection. (n = 3 independent experiments). Scale bar, 100 μm. Data are represented as mean ± SEM and statistical analysis was performed using two-tailed Student’s t-test (a, b, d, e, i–m) or one-way ANOVA with Dunnett’s multiple comparisons (g, h). Source data are provided as a Source Data file. AAV adeno-associated virus.