Fig. 6
Glycosaminoglycan-modified soluble–betaglycan (sBG) in ascites fluid is associated with decreased survival and TGF-β signaling responses for ovarian cancer patients. A, B Sol-BG concentration in AF samples plotted by A tumor stage, and B tumor type. Each data point represents a single patient sample. The blue line indicates the average sol-BG concentration in the plasma of healthy volunteers. C Correlation analysis of sol-BG concentration in the ascites fluid compared to patient overall survival in months. (n = 21) Spearman correlation analysis was performed, r = -0.4654, *p < 0.05; (p = 0.0335). D Kaplan–Meier survival plot of OVCA patients stratified into low (< 30th percentile) and high sol-BG (> 70th percentile) groups. A log-rank (Mantel-Cox) test was performed, p = 0.0236, median survival Low sol-BG = 47.2 months, High sol-BG = 22.0 months. E Autoradiograph of patient ascites fluid following [125 I]-TGF-β1 binding and crosslinking and immunoprecipitation using anti-BG antibody. F Western blot for SMAD2/3 phosphorylation in HEK293 cells treated with indicated OVCA patient ascites fluid. ns: nonspecific band G Correlation graph of SMAD2/3 phosphorylation in HEK293 cells, to the sol-BG concentration in the AF, used to treat HEK293 cells. Pearson correlation and simple linear regression were performed. (Pearson r = -0.7952, r2 = 0.3269, p = 0.0034). H Western blot of SMAD2/3 phosphorylation in HEK293 cells treated with OVCA patient ascites fluid with low sol-BG concentrations (< 400 pg/mL), with/without 1 ng/mL recombinant sol-BG and quantification of phospho-SMAD2/3 from (H) normalized to untreated samples and actin is plotted by Mean ± SEM, (n = 3), unpaired t-test between recombinant sBG untreated to treated samples. ns: nonspecific band *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. One-way ANOVA followed by unpaired t-test between sBG untreated (-) to treated ( +). (p = 0.0152 UAF8, p = 0.0002 UAF5, p = 0.2577 UAF7, p = 0.1576 UAF6, p = 0.0101 UAF3, p = 0.0027 UAF1)