Introduction
References |
N
| Adjuvant chemotherapy type | Cut-off point for BCL-2 immuno-positivity/overexpression | BCL-2 prognostic significance |
---|---|---|---|---|
van Slooten et al. (1996) | 423 | 202 pts: no adjuvant CT 221 pts: FAC | >75 % of cells with moderate or strong intensity | BCL-2 positivity: better DFS, lack of significance in multivariate analysis |
Mottolese et al. (2000) | 157 | 4 cycles of EC or 4 cycles of EC + lonidamine or 4 cycles of EC + G-CSF or 4 cycles of EC + lonidamine + G-CSF | >20 % of stained cells | In the entire cohort: no significance in lobular carcinomas: BCL-2 positivity: worse OS and DFS |
Yang et al. (2003) | 147 | All pts: FEC | BCL-2 positivity: better OS and DFS confirmed in multivariate analysis | |
Kröger et al. (2006) | 157 | 78 pts: 3 cycles of CMF 79 pts: 4 cycles of EC → CTM → stem cell support | >0 % of stained cells | BCL-2 positivity: better EFS, confirmed in multivariate analysis |
Lee et al. (2007) | 151 | AC → T | >0 % of stained cells | BCL-2 positivity: better OS and DFS confirmed in multivariate analysis |
Dumontet et al. (2010) | 1,342 | 663 pts: FAC 679 pts: TAC | >70 % of stained cells | BCL-2 positivity: better OS and DFS, lack of significance in multivariate analysis |
Abdel-Fatah et al. (2010) | 1,235 | 377 pts: no adjuvant treatment 382 pts: hormonal therapy 182 pts: CMF 32 pts: CMF + hormonal therapy 245 pts: anthracyclines | >10 % of stained cells | The entire patient cohort and separately patients treated with anthracyclines: BCL-2 positivity: better DFS and BCSS confirmed in multivariate analysis |
Kim et al. (2010) | 100 | 24 pts: no adjuvant treatment 30 pts: CMF 27 pts: doxorubicin containing CT 3 pts: others chemotherapy 16 pts: hormonal therapy only | >10 % of stained cells | No significance |
Materials and methods
Patients
Tumour samples
Immunohistochemistry
Antigen | Clone | Source | Dilution | Detection system | Criteria for IHC positivity/overexpression |
---|---|---|---|---|---|
ER | 6F11 | Novocastraa
| 1:50 | EnVisionb
| >10 % of tumour immunopositive cells |
PgR | 1A6 | Novocastraa
| 1:50 | EnVisionb
| >10 % of tumour immunopositive cells |
HER2 | – | DAKOb
| 1:250 | EnVisionb
| Strong complete uniform membrane staining in >30 % of tumour cells or complete membrane staining in >10 % of tumour cells confirmed by FISH |
CK 5/6 | D5/16 B4 | DAKOb
| 1:50 | EnVisionb
| >10 % of tumour immunopositive cells |
TOPOIIα | 3F6 | Novocastraa
| 1:30 | EnVisionb
| TOPOIIα labelling index (TOPOIIαLI—the percentage of TOPOIIα immunopositive tumour cells) >11.9 % (median value) |
Ki-67 | MIB-1 | DAKOb
| 1:75 | EnVisionb
| Ki-67 labelling index (Ki-67LI—the percentage of Ki-67 immunopositive tumour cells) >19.7 % (median value) |
CD34 | QBEnd 10 | DAKOb
| 1:50 | EnVisionb
| High microvessel density (MVD—mean number of microvessels per mm2 of tumour tissue) >210.0 vessels/mm2 (cut-off point found by minimal P value method) |
P53 | NCL-P53-1801 | Novocastraa
| 1:40 | EnVisionb
| P53 labelling (index (P53LI—the percentage of P53 immunopositive tumour cells) >10.0 % |
BCL-2 | 124 | DAKOb
| 1:40 | BrightVisionc
| Intense staining in all tumour cells (class 2) or heterogeneous staining within tumour area, regardless of the intensity (class 1) |
Statistical methods
Results
Patients
BCL-2 expression | |||||
---|---|---|---|---|---|
All (%)1
| 0 | 1 | 2 |
P valuea
| |
N (%)* |
N (%)* |
N (%)* | |||
All (%) | 141 (100) | 38 (27.0) | 70 (49.6) | 33 (23.4) | |
Age | |||||
<50 years | 47 (33.3) | 8 (17.0) | 29 (61.7) | 10 (21.3) | 0.092 |
≥50 years | 94 (66.7) | 30 (31.9) | 41 (43.6) | 23 (24.5) | |
Tumour size | |||||
T1 | 38 (27.0) | 9 (23.7) | 17 (44.7) | 12 (31.6) | 0.378 |
T2 | 103 (73.0) | 29 (28.1) | 53 (51.5) | 21 (20.4) | |
Nodal status | |||||
N1 | 108 (76.6) | 26 (24.1) | 56 (51.8) | 26 (24.1) | 0.376 |
N2 | 33 (23.4) | 12 (36.4) | 14 (42.4) | 7 (21.2) | |
Grade | |||||
G1 + G2 | 71 (50.4) | 10 (14.1) | 37 (52.1) | 24 (33.8) | 0.000 |
G3 | 70 (49.6) | 28 (40.0) | 33 (47.1) | 9 (12.9) | |
Oestrogen receptor status | |||||
Positive | 97 (68.8) | 11 (11.0) | 58 (58.0) | 31 (31.0) | 0.000 |
Negative | 44 (31.2) | 28 (63.6) | 14 (31.8) | 2 (4.6) | |
Progesterone receptor status | |||||
Positive | 93 (66.0) | 10 (10.7) | 53 (57.0) | 30 (32.3) | 0.000 |
Negative | 48 (34.0) | 28 (58.3) | 17 (35.4) | 3 (6.3) | |
Cytokeratin 5/6 expression | |||||
Positive | 30 (21.3) | 13 (43.3) | 15 (50.0) | 2 (6.7) | 0.015 |
Negative | 111 (78.7) | 25 (22.5) | 55 (49.6) | 31 (27.9) | |
HER2 status | |||||
Overexpressing | 54 (38.3) | 19 (35.2) | 27 (50.0) | 8 (14.8) | 0.084 |
Not overexpressing | 87 (61.7) | 19 (21.8) | 43 (49.4) | 25 (28.7) | |
TOPOIIαLIc
| |||||
≤11.9 % | 71 (50.4) | 16 (22.5) | 37 (52.1) | 18 (25.4) | 0.486 |
>11.9 % | 70 (49.6) | 22 (31.4) | 33 (47.1) | 15 (21.4) | |
Ki-67LIc
| |||||
≤19.7 % | 74 (52.5) | 15 (20.3) | 36 (48.6) | 23 (31.1) | 0.038 |
>19.7 % | 67 (47.5) | 23 (34.3) | 34 (50.7) | 10 (14.9) | |
MVDd
| |||||
≤210.0 vessels/mm2
| 96 (68.1) | 28 (29.1) | 47 (49.0) | 21 (21.9) | 0.645 |
>210.0 vessels/mm2
| 45 (31.9) | 10 (22.2) | 23 (51.1) | 12 (26.7) | |
P53LI | |||||
≤10.0 % | 100 (70.9) | 20 (20.0) | 55 (55.0) | 25 (25.0) | 0.014 |
>10.0 % | 41 (29.1) | 18 (43.9) | 15 (36.6) | 8 (19.5) | |
Breast cancer immunophenotypese
| |||||
LA | 42 (29.8) | 5 (11.9) | 19 (45.2) | 18 (42.9) | 0.000 |
LB HER2−
| 22 (15.6) | 0 (0.0) | 16 (72.7) | 6 (27.3) | |
LB HER2+
| 33 (23.4) | 5 (15.2) | 21 (63.6) | 7 (21.2) | |
HER2+
| 21 (14.9) | 14 (66.7) | 6 (28.5) | 1 (4.8) | |
TN | 23 (16.3) | 14 (60.9) | 8 (34.8) | 1 (4.3) |
BCL-2 expression, apoptosis level and correlations with clinicopathological features and other immunohistochemical markers
Univariate survival analysis
Response N (%) | HR | 95 % CI |
P value* | |
---|---|---|---|---|
Age | ||||
>50 years | 90/115 (78.3) | 1.080 | 0.534–2.189 | 0.830 |
≤50 years | 45/56 (80.4) | 1.000 | ||
Tumour size | ||||
T1 | 45/52 (86.5) | 1.000 | 0.846–4.374 | 0.109 |
T2 | 90/119 (75.6) | 1.924 | ||
Nodal status | ||||
N1 | 107/132 (81.1) | 1.000 | 0.783–3.214 | 0.206 |
N2 | 28/39 (71.8) | 1.587 | ||
Grade | ||||
G1 + G2 | 81/92 (88.0) | 1.000 | 1.508–6.198 | 0.001 |
G3 | 54/79 (68.4) | 3.057 | ||
Oestrogen receptor status | ||||
Positive | 102/123 (82.9) | 1.000 | 0.262–1.021 | 0.017 |
Negative | 33/48 (68.8) | 1.932 | ||
Progesterone receptor status | ||||
Positive | 98/119 (82.4) | 1.000 | 0.296–1.150 | 0.045 |
Negative | 37/52 (71.2) | 1.716 | ||
HER2 status | ||||
Overexpressing | 55/68 (80.9) | 1.000 | 0.523–1.984 | 0.750 |
Not overexpressing | 80/103 (77.7) | 1.019 | ||
Cytokeratin 5/6 | ||||
Positive | 23/35 (65.7) | 2.301 | 1.154–4.588 | 0.021 |
Negative | 112/136 (82.4) | 1.000 | ||
Ki-67LIa
| ||||
≤19.7 % | 74/90 (82.2) | 1.000 | 1.063–4.329 | 0.270 |
>19.7 % | 61/81 (75.3) | 2.196 | ||
TOPOIIαLIa
| ||||
≤11.9 % | 73/83 (88.0) | 1.000 | 1.376–5.886 | 0.003 |
>11.9 % | 62/88 (72.6) | 2.846 | ||
MVDb
| ||||
>210.0 vessels/mm2
| 52/53 (98.1) | 1.000 | 0.009–0.460 | 0.000 |
≤210.0 vessels/mm2
| 83/118 (71.3) | 15.724 | ||
P53 LI | ||||
>10.0 % | 35/50 (70.0) | 1.935 | 1.000–3.742 | 0.052 |
≤10 % | 100/121 (82.6) | 1.000 | ||
BCL-2 expression | ||||
Class 0 | 24/37 (64.9) | 2.873 | 0.156–0.776 | 0.005 |
Class 1 | 59/70 (84.3) | 1.302 | ||
Class 2 | 29/33 (87.9) | 1.000 | 0.246–2.401 | |
Breast cancer immunophenotypesd
| ||||
LA | 41/50 (80.8) | 1.000 | 0.350–2.164 | 0.081 |
LB HER2−
| 24/29 (79.6) | 1.084 | ||
LB HER2+
| 37/44 (84.0) | 1.094 | 0.386–2.410 | |
HER2+
| 18/24 (74.5) | 1.717 | 0.624–4.729 | |
TN | 15/24 (62.5) | 2.581 | 1.024–6.503 |
Multivariate analysis
HR | 95 % CI |
P value | |
---|---|---|---|
Grade | |||
1 + 2 | 1.000 | 0.611–4.176 | 0.329 |
3 | 1.610 | ||
Oestrogen receptor status | |||
Positive | 1.000 | 0.580–3.792 | 0.412 |
Negative | 1.484 | ||
Progesterone receptor status | |||
Positive | 1.000 | 0.355–1.748 | 0.560 |
Negative | 1.269 | ||
Cytokeratin 5/6 expression | |||
Positive | 1.801 | 0.782–4.144 | 0.169 |
Negative | 1.000 | ||
TOPOIIαLIa
| |||
≤11.9 % | 1.000 | 1.202–6.303 | 0.017 |
>11.9 % | 2.752 | ||
MVDb
| |||
>210.0 vessels/mm2
| 1.000 | 0.010–0.509 | 0.009 |
≤210.0 vessels/mm2
| 14.352 | ||
P53LI | |||
≤10.0 % | 1.000 | 0.536–2.679 | 0.662 |
>10.0 % | 1.198 | ||
BCL-2 expression | |||
Classes 1 + 2 | 1.000 | 0.153–0.699 | 0.004 |
Class 0 | 3.056 | ||
Breast cancer immunophenotypesd
| |||
LA + LB (HER2− and HER2+) | 1.000 | 0.322–2.886 | 0.947 |
HER2+ + TN | 1.038 |