Fig. 7: FBXO32-cyclin D1 axis limits the sensitivity of tumor to palbociclib.

a–e CCK-8, colony formation and trans-well assays were performed to measure the proliferation (a–c), migration (d) and invasion (e) of Huh7 and CFPAC-1 cells with stably expressing FBXO32 shRNA (sh-FBXO32) or negative control (sh-NC) by lentivirus infection in the presence of palbociclib (10 μM) or not. Scale bar, 100 μm. Statistical analysis was performed using one-way ANOVA with Tukey’s multiple comparisons, and data are represented as mean ± SEM (n = 3 independent experiments). f–i Huh7 and CFPAC-1 cells with stably expressing FBXO32 shRNA (sh-FBXO32) or negative control (sh-NC) were constructed through lentivirus infection, and then 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. Both sh-NC and sh-FBXO32 group mice were treated with palbociclib (100 mg/kg) or Vehicle (sodium lactate) daily by oral gavage administration for 14 days. The mice were sacrificed after the last administration. Representative images of primary liver (f) and pancreatic (h) tumors were presented, and the tumor volumes were monitored. Statistical analysis was performed using one-way ANOVA with Dunnett’s or Tukey’s multiple comparisons, and data are represented as mean ± SEM (For HCC, n = 7 biologically independent animals per group; For PDAC, n = 5 biologically independent animals per group). H&E staining was performed to determine the tumor tissue, and IHC staining was utilized to measure the protein levels of FBXO32 and cyclin D1 in HCC (g) and PDAC (i) tumor tissues. Scale bar, 50 μm. Source data are provided as a Source Data file.