Introduction
Materials and methods
Study population
Specimen characteristics
TMA construction
Assay methods
Antibody | Clone | Manufacturer | Concentration of antibody | Dilution |
---|---|---|---|---|
ER | SP1 | Cell marque | 33 mg/ml | 1:100 |
PR | 16 | Novocastra | 360 mg/l | 1:400 |
HER2 | CB11 | Novocastra | 3.9 g/l | 1:640 |
Ki67 | MIB1 | Dako | 35 mg/l | 1:100 |
CK5 | XM26 | Novocastra | 50 mg/l | 1:100 |
EGFR | 2-18C9 | Dako | Ready to use | No dilution |
Scoring and reporting
Classification of each marker
Classification of molecular subtypes
Statistical analyses
Ethics
Results
Description of the population
Luminal A | Luminal B (HER2−) | Luminal B (HER2+) | HER2 type | 5 Negative phenotype | Basal phenotype | Total | |
---|---|---|---|---|---|---|---|
Number (%) | 433 (47.6) | 249 (27.4) | 70 (7.7) | 60 (6.6) | 33 (3.6) | 64 (7) | 909 |
Mean age at diagnosis (SD) | 73.9 (9.9) | 71.9 (10.9) | 69 (11.4) | 67.3 (11.6) | 75.9 (11.1) | 71.7 (11.3) | 72.5 (10.7) |
Median years of follow-up after diagnosis (IQR) | 7.4 (9.3) | 7.0 (11.1) | 5.2 (12.5) | 3.2 (8.1) | 3.4 (8.5) | 5.1 (9.1) | 6.4 (10.0) |
Tumour grade (%) | |||||||
1 | 91 (21.0) | 20 (8.0) | 2 (2.9) | 0 | 0 | 4 (6.3) | 117 (12.9) |
2 | 297 (68.6) | 120 (48.2) | 33 (47.1) | 10 (16.7) | 21 (63.6) | 7 (10.9) | 488 (53.7) |
3 | 45 (10.4) | 109 (43.8) | 35 (50.0) | 50 (83.3) | 12 (36.4) | 53 (82.8) | 304 (33.4) |
Histopathological type (%) | |||||||
Ductal | 299 (69.1) | 182 (73.1) | 57 (81.4) | 47 (78.3) | 14 (42.4) | 37 (57.8) | 636 (70.0) |
Lobular | 68 (15.7) | 35 (14.1) | 6 (8.6) | 1 (1.7) | 12 (36.4) | 2 (3.1) | 124 (13.6) |
Tubular | 4 (0.9) | 0 | 0 | 0 | 0 | 0 | 4 (0,4) |
Mucinous | 31 (7.2) | 8 (3.2) | 3 (4.3) | 0 | 0 | 1 (1.6) | 43 (4.7) |
Papillary | 19 (4.4) | 7 (2.8) | 3 (4.3) | 1 (1.7) | 0 | 2 (3.1) | 32 (3.5) |
Medullary | 0 | 6 (2.4) | 0 | 6 (10.0) | 2 (6.1) | 7 (10.9) | 21 (2.3) |
Metaplastic | 0 | 1 (0.4) | 0 | 2 (3.3) | 1 (3.0) | 9 (14.1) | 13 (1.4) |
Other | 12 (2.8) | 10 (4.0) | 1 (1.4) | 3 (5.0) | 4 (12.1) | 6 (9.4) | 36 (4.0) |
Tumour sizea (%) | |||||||
<2 | 94 (21.7) | 50 (20.1) | 12 (17.1) | 4 (6.7) | 3 (9.1) | 6 (9.4) | 169 (18.6) |
2–5 | 193 (44.6) | 97 (39.0) | 27 (38.6) | 21 (35.0) | 18 (54.5) | 37 (57.8) | 393 (43.2) |
>5 | 35 (8.1) | 19 (7.6) | 7 (10.0) | 13 (21.7) | 2 (6.1) | 3 (4.7) | 79 (8.7) |
Uncertain | 111 (25.6) | 83 (33.3) | 24 (34.3) | 22 (36.7) | 10 (30.3) | 18 (28.1) | 268 (29.5) |
Lymph node invasiona
| |||||||
Yes | 129 (29.8) | 82 (32.9) | 25 (35.7) | 32 (53.3) | 15 (45.5) | 27 (42.2) | 310 (34.1) |
No (≥5 nodes or SNBb) | 142 (32.8) | 66 (26.5) | 24 (34.3) | 15 (25.0) | 5 (15.2) | 21 (32.8) | 273 (30.0) |
No (<5 nodes examined) | 123 (28.4) | 84 (33.7) | 20 (28.6) | 9 (15.0) | 10 (30.3) | 11 (17.2) | 257 (28.3) |
Uncertain | 39 (9.0) | 17 (6.8) | 1 (1.4) | 4 (6.7) | 3 (9.1) | 5 (7.8) | 69 (7.6) |
Stagec
| |||||||
I | 238 (55.0) | 123 (49.4) | 29 (41.4) | 23 (38.3) | 15 (45.5) | 27 (42.2) | 455 (50.1) |
II | 157 (36.3) | 90 (36.1) | 28 (40.0) | 27 (45.0) | 14 (42.4) | 30 (46.9) | 346 (38.1) |
III | 23 (5.3) | 17 (6.8) | 3 (4.3) | 7 (11.7) | 3 (9.1) | 4 (6.3) | 57 (6.3) |
IV | 13 (3.0) | 17 (6.8) | 8 (11.4) | 3 (5.0) | 1 (3.0) | 3 (4.7) | 45 (5.0) |
Unknown | 2 (0.5) | 2 (0.8) | 2 (2.9) | 0 | 0 | 0 | 6 (0.7) |
Histopathological characteristics
Marker | No. of positive (%) | Not possible to interpret |
---|---|---|
ER | 749 (82.4) | 2 (0.2 %) |
PR | 521 (57.3) | 1 (0.1 %) |
HER2 | 130 (14.3) | 0 |
Ki67 | 406 (44.7) | 1 (0.1 %) |
CK5 | 164 (18.0) | 0 |
EGFR | 64 (7.0) | 0 |
Distribution of molecular subtypes
Breast cancer-specific survival, molecular subtypes and histopathological grade
Risk of death from breast cancer
No. of cases | Deaths from breast cancer | HR 95 % CI unadjusted | HR 95 % CI adjusted for age | HR 95 % CI adjusted for stage | ||||
---|---|---|---|---|---|---|---|---|
Histopathological grade, follow-up first 5 years after diagnosis | ||||||||
1 | 117 | 13 | 1.00 | 1.00 | 1.00 | |||
2 | 488 | 101 | 1.97 | 1.11–3.51 | 1.95 | 1.09–3.48 | 1.47 | 0.82–2.64 |
3 | 304 | 110 | 3.80 | 2.14–6.75 | 3.74 | 2.10–6.66 | 3.12 | 1.75–5.55 |
909 | 224 | |||||||
Histopathological grade, follow-up from 5 years after diagnosisa
| ||||||||
1 | 78 | 18 | 1.00 | 1.00 | 1.00 | |||
2 | 291 | 83 | 1.37 | 0.82–2.29 | 1.29 | 0.77–2.16 | 1.21 | 0.72–2.02 |
3 | 153 | 34 | 0.98 | 0.55–1.74 | 0.94 | 0.53–1.67 | 0.90 | 0.51–1.60 |
522 | 135 | |||||||
Molecular subtype, follow-up first 5 years after diagnosis | ||||||||
Luminal A | 433 | 73 | 1.00 | 1.00 | 1.00 | |||
Luminal B (HER2−) | 249 | 56 | 1.42 | 1.01–2.02 | 1.42 | 1.00–2.02 | 1.29 | 0.92–1.84 |
Luminal B (HER2+) | 70 | 25 | 2.33 | 1.48–3.67 | 2.36 | 1.48–3.74 | 2.11 | 1.33–3.33 |
HER2 | 60 | 32 | 4.24 | 2.79–6.42 | 4.39 | 2.86–6.72 | 3.72 | 2.44–5.65 |
5 Negative phenotype | 33 | 15 | 3.34 | 1.91–5.82 | 3.18 | 1.81–5.61 | 3.17 | 1.81–5.53 |
Basal phenotype | 64 | 23 | 2.43 | 1.52–3.89 | 2.38 | 1.48–3.82 | 2.39 | 1.48–3.82 |
909 | 224 | |||||||
Molecular subtype, follow-up from 5 years after diagnosisa
| ||||||||
Luminal A | 271 | 69 | 1.00 | 1.00 | 1.00 | |||
Luminal B (HER2−) | 148 | 44 | 1.15 | 0.79–1.68 | 1.21 | 0.82–1.77 | 1.15 | 0.80–1.69 |
Luminal B (HER2+) | 36 | 10 | 0.81 | 0.41–1.57 | 0.88 | 0.44–1.73 | 0.92 | 0.46–1.83 |
HER2 | 23 | 4 | 0.66 | 0.24–1.80 | 0.71 | 0.26–1.96 | 0.66 | 0.24–1.82 |
5 Negative phenotype | 12 | 3 | 0.84 | 0.27–2.68 | 0.89 | 0.27–2.90 | 0.94 | 0.30–3.01 |
Basal phenotype | 32 | 5 | 0.58 | 0.23–1.43 | 0.62 | 0.25–1.55 | 0.58 | 0.23–1.46 |
522 | 135 |
Number of cases | Deaths from breast cancer | HR 95 % CI Unadjusted | HR 95 % CI Adjusted for age | HR 95 % CI Adjusted for stage | ||||
---|---|---|---|---|---|---|---|---|
Molecular subtype, follow-up first 5 years after diagnosis: grade 2 | ||||||||
Luminal A | 297 | 45 | 1.00 | 1.00 | 1.00 | |||
Luminal B (HER2−) | 120 | 24 | 1.45 | 0.88–2.38 | 1.50 | 0.91–2.48 | 1.33 | 0.81–2.19 |
Luminal B (HER2+) | 33 | 12 | 2.67 | 1.41–5.04 | 2.97 | 1.54–5.70 | 2.29 | 1.20–4.38 |
HER2 | 10 | 6 | 6.62 | 2.82–15.57 | 7.81 | 3.18–19.18 | 5.64 | 2.36–13.51 |
5 Negative Phenotype | 21 | 11 | 4.68 | 2.42–9.06 | 3.91 | 1.97–7.76 | 4.42 | 2.26–8.64 |
Basal Phenotype | 7 | 3 | 3.39 | 1.05–10.92 | 2.56 | 0.75–8.69 | 3.35 | 1.03–10.85 |
488 | 101 | |||||||
Molecular subtype, follow-up first 5 years after diagnosis: grade 3 | ||||||||
Luminal A | 45 | 16 | 1.00 | 1.00 | 1.00 | |||
Luminal B (HER2–) | 109 | 31 | 0.73 | 0.40–1.34 | 0.73 | 0.39–1.36 | 0.57 | 0.31–1.04 |
Luminal B (HER2+) | 35 | 13 | 1.00 | 0.48–2.09 | 0.96 | 0.45–2.05 | 0.85 | 0.41–1.79 |
HER2 | 50 | 26 | 1.60 | 0.86–2.99 | 1.60 | 0.84–3.05 | 1.21 | 0.64–2.29 |
5 Negative Phenotype | 12 | 4 | 0.90 | 0.30–2.70 | 0.87 | 0.28–2.64 | 0.94 | 0.31–2.82 |
Basal Phenotype | 53 | 20 | 1.07 | 0.55–2.06 | 1.07 | 0.54–2.11 | 0.86 | 0.44–1.68 |
304 | 110 |