Fig. 5 Proportions of EpCAM+ systemic tumor cells correlate with the clinical outcome of patients with MBC. Proportions of EpCAM+ systemic tumor cells.

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Fig. 5 Proportions of EpCAM+ systemic tumor cells correlate with the clinical outcome of patients with MBC. Proportions of EpCAM+ systemic tumor cells correlate with the clinical outcome of patients with MBC. (A) CTCs and DTCs were detected by SE-iFISH in pairs of blood and bone marrow samples from n = 34 patients with breast cancer. Shown are representative examples of aneuploid CTCs, DTCs, and cell clusters identified by iFISH (white arrows). DAPI (4′,6-diamidino-2-phenylindole), immunofluorescence (CD45, red; DAPI, blue; EpCAM, green), and FISH (CEP8, orange) are depicted as indicated. Shown are representative staining for each antigen. (B) Shown are numbers of detected CTCs and DTCs (top), numbers of cell clusters of CTCs and DTCs (middle), and percentages of EpCAM+ CTCs and DTCs in each patient (bottom). Paired CTC and DTC values are connected by solid lines. Wilcoxon signed-rank test, *P < 0.05 and ***P < 0.001. (C) Shown are proportions of EpCAM+ CTCs and DTCs in individual patients (n = 34) (left pie charts) and in overall numbers of CTCs (n = 845) and DTCs (n = 71,910) (right pie charts). EpCAM+, blue; EpCAM−, red. (D and H) Dot plots depict percentages of EpCAM+ CTCs and DTCs per patient (n = 34) stratified for distant metastases status M0 and M1 (without and with detectable organ metastasis, respectively) and for the absence or presence of lung metastases. (D) includes EpCAM+ diploid cells, whereas (H) excludes EpCAM+ diploid cells, as indicated. Mean percentage of EpCAM expression is indicated by a horizontal line. Mann-Whitney U test, *P < 0.05 and **P < 0.01. (E and I) Specificity and sensitivity of the EpCAM-positivity rate of DTCs to predict the 6-month survival rate are depicted as an ROC curve. AUC and P value are indicated. Sensitivity, true positives; 1-specificity, false positives. (E) includes EpCAM+ diploid cells, whereas (I) excludes EpCAM+ diploid cells. (F and J) Overall survival of patients with stages III and IV MBC (n = 34) was stratified according to the presence of EpCAM+ DTCs in the bone marrow with a cutoff of 20% (F) and 15% (J) (deduced from ROC analysis) and is depicted as Kaplan-Meier survival curves with P value. (F) includes EpCAM+ diploid cells, whereas (J) excludes EpCAM+ diploid cells, as indicated. Xiao Liu et al. Sci Adv 2019;5:eaav4275 Copyright © 2019 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).