Article Figures & Data
Figures
- Figure 1.
Gene expression levels of EMT-TFs (TWIST1, SNAIL1, ZEB1, and TG2) and HER2 in the CD45+ and CD326+ cell fractions. The GAPDH gene was used to normalize gene expression levels, and the relative expression of each gene was calculated using the equation
, where
. Samples were considered positive if the gene expression of was higher than the mean of that in the CD45+ cell fraction + 3 SD (cut-off line). The medians among fractions were compared using the Mann–Whitney test. - Figure 2.
Gene expression levels of EMT-TFs (TWIST1, SNAIL1, ZEB1, and TG2) and HER2 in the CD45+ and CD45− cell fractions. The GAPDH gene was used to normalize gene expression levels, and the relative expression of each gene was calculated using the equation
, where
. Samples were considered positive if the gene expression of was equal to or higher than 2 (cut-off line). - Figure 3.
A, flow cytometric gating strategy to identify the CSC populations (ALDEFLUOR+ CD44+CD24lo/CD45− and ALDEFLUOR+ CD133+/CD45−). SSC, side scatter. B, we compared the percentage of stem cell marker–positive cells, ALDEFLUOR+ CD44+CD24lo/CD45− on the left and ALDEFLUOR+ CD133+/CD45− on the right, in the CD45− cell fractions with and without amplified EMT-TFs. The groups were compared using the Mann–Whitney test as follows: at least one amplified EMT-TF versus none, trastuzumab-naïve (group 1) versus trastuzumab-treated (group 2).
- Figure 4.
PFS (A) and OS (B) Kaplan–Meier curves for all 17 patients according to EpCAM transcript expression in the CD326+ cell fraction. EpCAM high expression (solid line; n = 8); EpCAM low expression (dashed line; n = 9); time was measured from the basal blood draw to the death for OS and to radiological evidence of disease progression (PFS).
Tables
- Table 1.
Patient characteristics
Characteristic n (%) Overall 28 (100) Age [median (range)], y 52 (35–72) ER/PR+ 15 (53.6) ER/PR− 13 (46.4) First-line 10 (35.7) ≥Second-line 18 (64.3) CTC = 0 12 (42.9) CTC = 1–4 7 (25) CTCs ≥ 5 3 (10.7) CellSearch not conducted 6 (21.4) Abbreviations: ER, estrogen receptor; PR, progesterone receptor.
- Table 2.
EMT-TFs expression according to CTC count and treatment
Subgroups Data available (no. of patients) EMT-TF+ CD326+, n/N (%) EMT-TF+ CD45−, n/N (%) No. of CTCs 0 12 4/6 (66.7) 6/12 (50) 1–4 7 3/3 (100) 3/7 (42.9) ≥5 3 1/1 (100) 3/3 (100) Trastuzumab-naïve (group 1) 9 5/6 (83.3) 8/9 (88.9) Trastuzumab-treated (group 2) 18 10/11 (90.9) 8/18 (44.4)
Supplementary Data
Files in this Data Supplement:
- Supplementary Figure Legend - PDF file - 86K
- Supplementary Figure 1 - PDF file - 64K, Gene expression levels of EMT-TFs in the tested cell lines (HMLE, HMLE-TWIST, MCF-7, BT-474, SKBR-3, MDA-MB-231)
Volume 11, Issue 11
