Fig. 8

Pentose phosphate pathway (PPP) is associated with FBP1 and Sunitinib resistance in ccRCC. A) Glucose uptake and lactate secretion measured in 786O cells with FBP1 overexpression (AvFBP1) or control (AvCON) under hypoxia and normoxia; B) Relative firefly-luciferase activity of hypoxia response element (HRE) in 786O cells under hypoxia and normoxia in 786O cell with FBP1 overexpression, knockdown (shFBP1) or control; C) mRNA expression of target genes of HIF1A in 786O cells with FBP1 overexpression or control; D) Reproduced from in GEO dataset, shown was violin plot of PFKFB4 expression level in pretreat and Sunitinib (Sun)-resistant ccRCC, Student’s t-test; E) Proliferation detected by crystal violet assay in 3 ccRCC cell lines with different sensitivity to Sun harvested on day 3, with short exposure denoting sensitive cells being exposed to 30 µM of Sun for 72 h, two-way ANOVA; F) Shifting of sigmoidal dose-response fitting curve of Sun applied to 2 ccRCC cell lines with lentiviral overexpression (Lv) or knockdown (sh) of target genes; G) Proliferation detected using crystal violet at 72 h of treatment with 10 µM of 5MPN (M10) or combined with different doses (µM) of Sun (S30 to S1) in 2 ccRCC cell lines with different Sun sensitivity profile, all normalized to control (CON); Metabolic analysis in 786O cells with short Sunitinib exposure, Sunitinib –sensitive and -resistant 786O cells, shown were H) Glucose uptake and lactate secretion; I) Intact cellular respiration measured using live cell real-time metabolism monitoring under basal conditions or in the presence of FCCP; J) Glucose oxidation measured by ¹⁴ C-CO₂ production following 3-hour labeling with D[U-14 C]glucose; K) Western blotting of select Sunitinib targets and PPP genes. (All in vitro assays performed in triplicates and at least 3 biological replicates; ns = not significant; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001)