Figure 3
From: Endo180 modulation by bisphosphonates and diagnostic accuracy in metastatic breast cancer
Endo180 levels are elevated in the plasma of patients with metastatic compared with early BCa. Plasma samples from BCa patients (n=88) were resolved on 8% w/v SDS polyacrylamide gels, transferred to polyvinylidene fluoride membranes and immunoblot analysis was performed using 1 mg ml−1 mouse anti-human Endo180 monoclonal 39.10 antibody and 1/4000 dilution of horse radish peroxidase-conjugated goat anti-mouse IgG (a representative blot is presented in Supplementary Figure 1). Relative plasma Endo180 levels were calculated from the mean measurement obtained from three analytical repeats. Plasma from the same early BCa patient was used as a reference control (relative Endo180 level=1.0) and the normalisation of densitometric measurements on all analytical gels. CA 15-3 antigen concentrations (U ml−1) were measured using a non-competitive two-step immunoassay in plasma samples (n=86a) collected in parallel. The box and whiskers plots shown depict the total range, interquartile distribution, outliers (open or closed circles) and median value (horizontal line) of relative plasma Endo180 levels (A, C) and plasma concentrations of CA 15-3 antigen (Log10 [U ml−1]) (B, D). (A) The graph compares relative plasma Endo180 levels in cases of early BCa (n=29) and metastatic BCa (n=59). (B) The graph compares plasma concentrations of CA 15-3 antigen (Log [U ml−1]) in cases of early BCa (n=27a) and metastatic BCa (n=59). (C) The graph compares relative plasma Endo180 levels in cases of early BCa (n=29) and recurrent locoregional disease (n=5) or metastasis in visceral tissue alone (n=12), bone alone (n=14) or bone with dissemination in lung (n=5), liver (n=8) or two or more visceral organs (n=15). (D) The graph compares plasma concentrations of CA 15-3 antigen (Log10 [U ml−1]) in cases of early BCa (n=27*) and metastatic sites as described above (C). The P-values shown were determined using Mann–Whitney U-test for comparison between cases of early BCa and metastatic BCa (A, B); or Kruskal–Wallis one-way analysis of variance for comparison between early BCa and recurrent/metastatic BCa stratified according to affected tissue sites (C, D), and confirmed by Bonferroni adjustment (P=0.05/n, n=7). aData for CA 15-3 antigen were missing for 2 out of the 29 early BCa patients included in the study.
