Fig 2: Androgen-independent LNCaP cells exhibits enhanced glycolysis. A, The basal glucose utilization in LNCaP and LNCaP-AI cells. B, The basal lactate production in LNCaP and LNCaP-AI cells. C, Comparison of ECAR and OCR status in LNCaP and LNCaP-AI cells. D, Real-time qPCR analysis of glucose transporter and key glycolytic genes in LNCaP and LNCaP-AI cells. E, Western blotting analysis of the effect of PRKAR2B knockdown efficiency in LNCaP-AI cells. (F-I) The effect of PRKAR2B knockdown on the glucose utilization (F), lactate production (G), ECAR/OCR (H), and expression of glycolytic components (I) in LNCaP-AI cells. (J) Correlation analysis of the link between PRKAR2B expression and the expression level of glucose transporter and key glycolytic genes in prostate cancer tissues (n = 492); data were obtained from the TCGA cohort. *P < .05; **P < .01

Fig 3: PRKAR2B overexpression promotes the Warburg effect in prostate cancer in vitro. A, Western blotting analysis of the overexpression efficiency of PRKAR2B in LNCaP cells. B-E, The effect of PRKAR2B overexpression on the glucose utilization (B), lactate production (C), extracellular acidification rate (D), oxygen consumption rate (E), and expression of glycolytic components (F) in LNCaP cells. SLC2A1, solute carrier family 2 member 1; PFKP, phosphofructokinase platelet; PKM, pyruvate kinase muscle isozyme; LDHA, lactate dehydrogenase A. *P < .05; **P < .01

Fig 4: The growth-promoting role of PRKAR2B in prostate cancer is glycolysis-dependent. A, In vivo growth assay of that sh-Ctrl and sh-PRKAR2B DU145 cells. B, In vivo growth assay of that ov-vector and PRKAR2B-overexressing LNCaP cells. C, Plate clone formation assay showed the effect of PRKAR2B on LNCaP cell proliferation in the presence or absence of 5 mmol/L 2-DG. D, Plate clone formation assay showed the effect of PRKAR2B on LNCaP cell proliferation in the culture medium containing 25 mmol/L glucose or 25 mmol/L galactose. E, Measurement of the effect of PRKAR2B overexpression on LNCaP cell proliferation in the presence or absence of HIF-1a knockdown. F, Real-time qPCR analysis of PRKAR2B expression in DU145 and PC3 cells after 2-DG treatment. G, Real-time qPCR analysis of PRKAR2B expression in DU145 and PC3 cells after galactose treatment. *P < .05; **P < .01

Fig 5: PRKAR2B regulates HIF1a expression in prostate cancer. (A-B) Correlation analysis of the link between PRKAR2B and HIF1A expression in prostate cancer tissues; data were obtained from the TCGA cohort (A) and Ren Ji cohort (B). (C) Western blotting analysis of the effect of PRKAR2B knockdown on the HIF1A protein level in DU145 cells. (D) Western blotting analysis of the effect of PRKAR2B overexpression on the HIF1A protein level in LNCaP cells. (E) Western blotting analysis of HIF1A protein level in LNCaP and LNCaP-AI cells. (F) Western blotting analysis of the effect of PRKAR2B knockdown on the HIF1A protein level in LNCaP-AI cells. (G) Effects of PRKAR2B knockdown or overexpression on the HIF-1a activity. (H) Western blotting analysis of HIF1A protein level in DU145 and PC3 cells upon H89 (50 µmol/L) treatment for 24 h. (I) Determining the effect of H89 (50 µmol/L) treatment on the HIF-1a activity in DU145 and PC3 cells. *P < .05; **P < .01