Fig. 7
From: ADP-dependent glucokinase controls metabolic fitness in prostate cancer progression
ADPGK binds with ALDOC, activating the AMPK pathway. a Protein–protein interaction network analysis from quantitative proteomics. b Correlation analysis between ADPGK and ALDOC expression from cBioPortal (data from Neuroendocrine Prostate Cancer Multi-Institute). c Immunofluorescence staining for His-tag (blue) and ALDOC (red) primary antibodies in PC3-ADPGK-His-tag cells. The mean colocalization Pearson’s correlation coefficient was 0.78 (n = 3). Scale bar = 20 μm. d Co-immunoprecipitation experiments showed the interactions between ADPGK and ALDOC (5 μg of His-tag and IgG IP antibody). e ALDOC expression in 22Rv1 cells after ADPGK knockdown, as detected by an immunofluorescence assay. The mean immunofluorescence intensity was evaluated by ImageJ software (n = 5). Scale bar = 20 μm. f Effect of ADPGK knockdown on ALDOC expression in 22Rv1 cells detected by Western blotting. Effect of ADPGK overexpression on ALDOC expression in PC3 (g) and 22Rv1 (h) cells detected by Western blotting. i siRNA-mediated ALDOC knockdown detected by Western blotting. ECAR test in LNCaP (j) and 22Rv1 (k) cells after ADPGK or ALDOC knockdown (n = 3). l Effect of ALDOC knockdown on ALDOC, AMPK and p-AMPK expression in 22Rv1 and LNCaP cells. m Effect of ADPGK overexpression and siALDOC#2 on ADPGK, ALDOC, AMPK and p-AMPK expression in PC3 cells. n ALDOC expression status in clinical samples assessed by immunohistochemistry (IHC). Scale bar = 50 μm. o Representative IHC quantification of ADPGK and ALDOC expression status in PCa tissues (χ2 test). p Kaplan‒Meier curves showing the association of ALDOC expression with biochemical recurrence (BCR) in 45 patients. q Kaplan‒Meier curves showing survival outcomes among PCa patients stratified into three groups: ADPGKhigh/ALDOChigh group (n = 16), ADPGKlow/ALDOChigh + ADPGKhigh/ALDOClow group (n = 17), and ADPGKlow/ALDOClow group (n = 12). Data are presented as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. PCa prostate cancer, BPH benign prostate hyperplasia, ADPGK ADP-dependent glucokinase, ALDOC aldolase C, AMPK AMP-activated protein kinase, NC negative control, OE overexpression, ECAR extracellular acidification rate, 2-DG 2-deoxy-glucose, SD standard deviation