TABLE 1.
pPCR analysis of breast cancer cell lines overexpressing HS sulfotransferases.
| Genes | MCF-7 |
MDA-MB-231 |
||||||||
| Control | HS2ST1 | P | HS3ST2 | P | Control | HS2ST1 | P | HS3ST2 | P | |
| HS related | ||||||||||
| HS2ST1 (n ≥ 10) | 1.00 ± 0.07 | 24.89 ± 16.56 | <0.001 | 1.06 ± 0.23 | 0.51 | 1.01 ± 0.11 | 37.42 ± 18.47 | <0.0001 | 1.10 ± 0.18 | 0.15 |
| HS3ST2 (n ≥ 6)* | – | – | n.a. | +++ | n.a. | – | – | n.a. | +++ | n.a. |
| Sulf1 (n ≥ 8) | 1.00 ± 0.20 | 15.46 ± 12.02 | <0.05 | 7.52 ± 4.53 | <0.01 | 1.02 ± 0.23 | 3.88 ± 1.59 | <0.01 | 1.65 ± 0.51 | <0.01 |
| Sulf2 (n ≥ 8) | 1.01 ± 0.12 | 0.84 ± 0.19 | 0.06 | 0.72 ± 0.24 | <0.05 | 1.08 ± 0.42 | 2.62 ± 2.66 | 0.15 | 1.87 ± 1.42 | 0.17 |
| HPSE (n ≥ 10) | 1.03 ± 0.28 | 1.97 ± 0.91 | <0.001 | 1.50 ± 0.92 | <0.05 | 1.02 ± 0.20 | 0.57 ± 0.31 | <0.0001 | 1.02 ± 0.31 | 0.97 |
| Syndecans | ||||||||||
| Sdc-1 (n ≥ 6) | 1.02 ± 0.21 | 1.03 ± 0.18 | 0.93 | 1.03 ± 0.24 | 0.91 | 1.03 ± 0.27 | 0.75 ± 0.54 | 0.28 | 0.75 ± 0.60 | 0.33 |
| Sdc-2 (n ≥ 6) | 1.03 ± 0.28 | 1.04 ± 0.19 | 0.92 | 0.84 ± 0.26 | 0.31 | 1.01 ± 0.10 | 2.04 ± 0.67 | <0.04 | 1.19 ± 0.31 | 0.86 |
| Sdc-3 (n ≥ 3) | 1.00 ± 0.09 | 1.09 ± 0.18 | 0.53 | 0.84 ± 0.07 | 0.07 | 1.01 ± 0.13 | 1.14 ± 0.58 | 0.74 | 0.79 ± 0.05 | 0.09 |
| Sdc-4 (n ≥ 8) | 1.02 ± 0.23 | 1.80 ± 0.66 | <0.001 | 2.06 ± 0.68 | <0.01 | 1.01 ± 0.16 | 1.54 ± 0.72 | 0.07 | 1.25 ± 0.70 | 0.37 |
| EMT | ||||||||||
| E-Cadherin (n ≥ 9) | 1.01 ± 0.06 | 0.79 ± 0.20 | <0.01 | 0.88 ± 0.37 | 0.28 | 1.00 ± 0.04 | 1.24 ± 0.85 | 0.38 | 1.43 ± 0.67 | 0.09 |
| N-Cadherin (n ≥ 7) | 1.15 ± 0.60 | 1.00 ± 0.50 | 0.60 | 0.96 ± 0.41 | 0.48 | 1.01 ± 0.12 | 1.23 ± 0.19 | <0.05 | 0.82 ± 0.25 | 0.09 |
| β-catenin (n ≥ 7) | 1.01 ± 0.18 | 1.42 ± 0.33 | <0.05 | 1.21 ± 0.29 | 0.13 | 1.01 ± 0.12 | 0.81 ± 0.32 | 0.14 | 1.15 ± 0.46 | 0.42 |
| ZEB1 (n ≥ 8) | 1.08 ± 0.46 | 1.34 ± 0.77 | 0.39 | 1.42 ± 0.63 | 0.23 | 1.00 ± 0.07 | 1.12 ± 0.29 | 0.27 | 0.80 ± 0.22 | <0.05 |
| ZEB2 (n ≥ 7) | 0.93 ± 0.36 | 2.74 ± 4.02 | 0.28 | 2.38 ± 2.21 | 0.17 | 1.01 ± 0.17 | 1.51 ± 0.77 | 0.11 | 1.23 ± 0.34 | 0.14 |
| Vimentin (n ≥ 9) | 1.01 ± 0.05 | 0.62 ± 0.34 | <0.01 | 0.79 ± 0.45 | 0.13 | 1.00 ± 0.01 | 1.02 ± 0.08 | 0.42 | 0.87 ± 0.06 | <0.0001 |
| Snail1 (n ≥ 9) | 1.02 ± 0.24 | 0.58 ± 0.46 | <0.001 | 0.90 ± 0.19 | <0.001 | 1.00 ± 0.08 | 0.67 ± 0.21 | <0.01 | 1.41 ± 0.41 | <0.05 |
| Snail2 (n ≥ 16) | 1.08 ± 0.33 | 0.64 ± 0.32 | <0.05 | 0.87 ± 0.50 | 0.54 | 1.01 ± 0.13 | 0.68 ± 0.22 | <0.01 | 1.32 ± 0.34 | <0.05 |
| Twist (n ≥ 9) | 1.00 ± 0.06 | 0.08 ± 0.19 | <0.0001 | 0.11 ± 0.25 | <0.0001 | 1.03 ± 0.27 | 3.05 ± 3.81 | 0.15 | 2.56 ± 3.70 | 0.25 |
| Notch signaling | ||||||||||
| Notch-1 (n ≥ 5) | 1.03 ± 0.27 | 1.09 ± 0.17 | 0.67 | 0.92 ± 0.29 | 0.57 | 1.06 ± 0.41 | 1.33 ± 1.18 | 0.51 | 0.98 ± 0.27 | 0.58 |
| Notch-2 (n ≥ 3) | 1.00 ± 0.01 | 5.78 ± 6.25 | 0.32 | 4.62 ± 4.44 | 0.29 | 1.09 ± 0.50 | 3.19 ± 1.57 | <0.001 | 6.81 ± 5.72 | <0.05 |
| Notch-3 (n ≥ 5) | 1.02 ± 0.23 | 1.69 ± 0.10 | <0.01 | 1.54 ± 0.27 | <0.05 | 1.13 ± 0.62 | 1.59 ± 1.17 | 0.31 | 2.56 ± 2.52 | 0.11 |
| Notch-4** (n ≥ 5) | 1.35 ± 1.19 | 0.39 ± 0.39 | 0.08 | 1.21 ± 1.40 | 0.84 | 0.98 ± 0.34 | 0.65 ± 0.44 | 0.09 | 0.86 ± 0.52 | 0.59 |
| Numb (n ≥ 12) | 1.00 ± 0.11 | 1.20 ± 0.33 | 0.06 | 1.16 ± 0.18 | <0.01 | 1.02 ± 0.21 | 1.74 ± 1.07 | <0.05 | 1.04 ± 0,41 | 0.87 |
| DLL1 (n ≥ 3) | 1.00 ± 0.09 | 0.93 ± 0.33 | 0.75 | 1.27 ± 0.28 | <0.05 | 1.01 ± 0.08 | 1.23 ± 0.14 | <0.0001 | n.d. | n.d. |
| DLL3 (n ≥ 8) | 1.07 ± 0.42 | 1.43 ± 2.21 | 0.66 | 0.88 ± 0.96 | 0.52 | 1.04 ± 0.32 | 0.94 ± 0.48 | 0.61 | 0.82 ± 0.45 | 0.25 |
| DLL4 (n ≥ 8) | 1.03 ± 0.28 | 0.83 ± 0.40 | 0.26 | 0.70 ± 0.32 | <0.05 | 1.03 ± 0.28 | 1.77 ± 0.72 | <0.05 | 0.93 ± 0.18 | 0.43 |
| Hes1 (n ≥ 9) | 1.04 ± 0.29 | 1.05 ± 0.41 | 0.69 | 0.80 ± 0.31 | 0.05 | 1.02 ± 0.24 | 2.04 ± 1.18 | <0.05 | 1.73 ± 0.73 | <0.05 |
| Hes2 (n ≥ 16) | 1.03 ± 0.25 | 0.72 ± 0.18 | <0.0001 | 0.87 ± 0.40 | 0.18 | 1.03 ± 0.18 | 1.88 ± 0.80 | <0.0001 | 1.01 ± 0.47 | 0.91 |
| Hey1 (n ≥ 11) | 1.02 ± 0.10 | 1.30 ± 0.29 | <0.001 | 1.01 ± 0.38 | 0.94 | 1.00 ± 0.07 | 0.87 ± 0.30 | 0.17 | 1.01 ± 0.13 | 0.88 |
| Hey2 (n ≥ 17) | 1.15 ± 0.69 | 5.58 ± 7.96 | <0.05 | 3.77 ± 4.18 | p < 0.05 | 1.07 ± 0.44 | 1.10 ± 0.71 | 0.86 | 1.66 ± 1.08 | p < 0.05 |
| Jag1 (n ≥ 15) | 1.10 ± 0.50 | 2.60 ± 1.28 | <0.001 | 2.41 ± 0.94 | <0.001 | 1.02 ± 0.23 | 1.45 ± 0.57 | <0.05 | 1.18 ± 0.78 | 0.46 |
| Wnt signaling | ||||||||||
| Wnt1 (n ≥ 8) | – | – | n.a. | – | n.a. | 1.01 ± 0.46 | 1.07 ± 0.65 | 0.82 | 2.48 ± 1.11 | <0.01 |
| Wnt3a (n ≥ 8) | 1.01 ± 0.18 | 1.61 ± 1.42 | 0.20 | 1.83 ± 1.62 | 0.13 | 1.07 ± 0.42 | 1.18 ± 0.77 | 0.71 | 1.32 ± 0.90 | 0.48 |
| Wnt5a (n ≥ 8) | 1.38 ± 1.15 | 1.39 ± 0.68 | 0.99 | 1.58 ± 0.92 | 0.72 | 1.00 ± 0.04 | 0.49 ± 0.11 | <0.0001 | 0.29 ± 0.11 | <0.0001 |
| FZD7 (n ≥ 8) | 1.02 ± 0.24 | 1.12 ± 0.40 | 0.57 | 1.13 ± 0.58 | 0.66 | 1.00 ± 0.08 | 1.19 ± 0.14 | <0.01 | 1.73 ± 0.60 | <0.05 |
| SFRP1 (n ≥ 8) | 1.20 ± 0.74 | 1.40 ± 1.69 | 0.73 | 1.16 ± 0.81 | 0.90 | 1.03 ± 0.27 | 1.58 ± 0.66 | 0.06 | 0.89 ± 0.54 | 0.52 |
| TCF7L1 (n ≥ 8) | 1.09 ± 0.31 | 0.65 ± 0.40 | <0.001 | 0.58 ± 0.24 | <0.001 | 1.02 ± 0.23 | 4.23 ± 3.10 | <0.05 | 1.63 ± 0.40 | <0.01 |
| Others | ||||||||||
| CD133 (n ≥ 7) | 0.95 ± 0.36 | 1.16 ± 0.95 | 0.58 | 1.43 ± 0.39 | <0.05 | 1.02 ± 0.22 | 1.46 ± 0.41 | <0.05 | 0.67 ± 0.14 | <0.01 |
| Myc (n ≥ 9) | 1.01 ± 0.18 | 0.49 ± 0.21 | <0.0001 | 0.56 ± 0.20 | <0.01 | 1.01 ± 0.18 | 1.46 ± 0.43 | 0.06 | 0.71 ± 0.30 | <0.01 |
| CyclinD1 (n ≥ 8) | 1.02 ± 0.19 | 0.62 ± 0.18 | <0.001 | 0.50 ± 0.22 | <0.001 | 1.00 ± 0.06 | 1.23 ± 0.26 | <0.05 | 1.00 ± 0.29 | 0.96 |
Gene expression data are displayed as fold-change value (2–ΔΔCt) relative to vector control cells (mean ± SD). Statistically significant data are displayed in bold typing. *HS2ST3 is not expressed in MCF-7 and MDA-MB-231 cells. – denotes absence of expression. +++, denotes high expression due to stable transfection. **, Notch4 expression data were at the limit of detection (Ct values > 32). n.d., not determined. n.a., not applicable.