Fig 2: NRF2 mediates the effects of oxPAPC on glycolysis and proliferation. (A) Glycolysis stress test (mean ± SEM, n=5), glycolytic rate (mean ± SD, n=5) and glycolytic capacity (mean ± SD, n=5) in oxPAPC and control treated endothelial cells. (B) PFKFB3 expression in oxPAPC-treated cells and representative Western blot of PFKFB3 of oxPAPC (8h) and control treated cells. Fold changes are calculated against respective control treatments in each time point (n=6). (C) Proliferation assay for control and oxPAPC-treated cells. Fold change is calculated against control cells (n=6). (D) Expression of VEGFA and MYC in control and oxPAPC treated (12h) cells. Fold changes are calculated against VEGFA control values (n=3). (E) Representative Western blot for MYC in control and oxPAPC treated (12h) cells. (F) Overview of the suggested mechanism (for B-D: mean ± SD, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001).

Fig 3: PFKFB3 knockdown suppressed the aggressive behavior of HCC cells in vitro. (A-D) HCC-LM3 and PLC cells were infected with si-Control or si-PFKFB3. The cells were harvested for CCK8 (A-B), colony formation (C), and Transwell migration assays (D) after 48 h of culture. Each RNAi group was compared with the control group. Data are presented as mean ± SD. * P < 0.05; ** P < 0.01; *** P < 0.001.

Fig 4: Sorafenib decreased the tumor volume, and the administration of sorafenib with a median dosage (20 mg/kg) increased the intrahepatic metastatic potential and lung metastatic potential of liver cancer cells; sorafenib with a high dosage (40 mg/kg) inhibited intrahepatic metastasis and lung metastasis. PFKFB3 knockout attenuated the pro-invasive effect of sorafenib treatment in vivo. (A) Sorafenib decreased the tumor volume in the HCCLM3‑wt and HCCLM3-PFKFB3(-) groups (both * P < 0.05, respectively). (B and C) Sorafenib with a median dosage (20 mg/kg) increased the number of IHM in the HCCLM3‑wt sorafenib group compared with that in the HCCLM3-wt control group (* P < 0.05), while sorafenib with a high dosage (40 mg/kg) did not increase the number of IHMs in the HCCLM3-wt group; sorafenib did not increase the number of IHMs in the HCCLM3-PFKFB3(-) sorafenib group compared with that in the HCCLM3-PFKFB3(-) control group. The number of IHMs in the HCCLM3-wt control was not higher than that in the HCCLM3-PFKFB3(-) control (P > 0.05). (D and E) Sorafenib increased the number of lung metastases in the HCCLM3‑wt sorafenib group compared with that in the HCCLM3‑wt control, while sorafenib did not increase the number of lung metastases in the HCCLM3-PFKFB3(-) sorafenib group compared with that in the HCCLM3-PFKFB3(-) control. (** P < 0.01). The number of lung metastases in the HCCLM3-wt control was higher than that in the HCCLM3-PFKFB3(-) control.

Fig 5: The p65 activation in CAC. (a) The phosphorylation of p65 detected in PFKFB3 overexpression and knockdown. (b) The nuclear translocation of p65 detected in PFKFB3 overexpression and knockdown; scar bar = 20 μm. (c)-(d) The expression of IL-1β and TNF-α level by PFKFB3 overexpression and p65 knockdown in IECs. (e) The proliferation of CRC by PFKFB3 overexpression and p65 knockdown in IECs. (f) The invasion of CRC by PFKFB3 overexpression and p65 knockdown in IECs; scar bar = 50 μm. ∗P < 0.05, ∗∗p < 0.01.