Introduction
Materials and methods
Study patients
Tissue arrays and immunohistochemistry
Antibody | Dilution | Source/clone | Pretreatment/solution | Cut-off |
---|---|---|---|---|
Hormone receptor | ||||
ERa
| 1:200 | DAKO, ID5 | Microwave, citrate pH 6.0 | ≥1 % |
DNA damage repair molecules | ||||
PARP1 (Cleaved) | 1:1,000 | Abnova, A6.4.12 | Microwave, citrate pH 6.0 | >200 H-score |
PARP1 (Non-cleaved) | 1:1,000 | BD Pharmingen, 7D3-6 | Microwave, citrate pH 6.0 | >10 H-score |
RAD51 | 1:70 | Abcam, mouse polyclonal | Microwave, citrate pH 6.0 | |
BRCA1 | 1:150 | Oncogene research, MS110 | Microwave, citrate pH 6.0 | >100 H-score |
Ku70/Ku80 | 1:2,500 | Abcam, mouse monoclonal | Microwave, citrate pH 6.0 | >90 H-score |
PIASγ | 1:50 | Abcam, anti E3 (ab61405) | Microwave, citrate pH 6.0 | >160 H-score |
BRCA1 transcriptional suppressor molecule | ||||
ID4 | 1:300 | Abcam, ab 20988- 100 | Microwave, citrate pH 6.0 | >12 H-score |
EGFR family members | ||||
EGFR | 1:30 | Zymed, 31G7 | Proteinase K | >10 % |
c-erbB-2 (HER2) | 1:250 | DAKO, c-erbB-2 | None | 3+ (2+ with CISH) |
Cell cycle, proliferation and apoptosis related proteins | ||||
p53 | 1:50 | Novocstra, DO7 | Microwave, citrate pH 6.0 | >10 % |
MDM2 | 1:50 | Novocastra, IB10 | Microwave, citrate pH 6.0 | ≥10 % |
Ki-67/clone MIB1a
| 1:100 | DAKO, M7240 | Microwave, citrate pH 6.0 | >10 % |
Bcl2 | 1:100 | DAKO, 124 | Microwave, citrate pH 6.0 | >10 % |
CHK1 | 1:150 | Abcam, rabbit polyclonal | Microwave, citrate pH 6.0 | >30 H-score |
CK5/6 | 1:100 | Boehringer biochemica, D5/16134 | Microwave, citrate pH 6.0 | >10 % |
CK14 | 1:100 | Novocastra, LL002 | Microwave, citrate pH 6.0 | >10 % |
CK17 | 1:20 | Abcam, E3 | Microwave, citrate pH 6.0 | >10 % |
Evaluation of immunohistochemical staining
Statistical analysis
Results
PARP1 non-cleaved n (%) | PARP1 cleaved n (%) | ||||||
---|---|---|---|---|---|---|---|
Negative | Positive |
p value | Negative | Positive |
p value | ||
Unselected sporadic series | 552 (51.3) | 524 (48.7) | N/A | 194 (15.4) | 1075 (84.6) | N/A | |
BRCA1-mutated tumours | 2 (4.7) | 41 (95.3) | 5 (10.4) | 38 (79.2) | |||
Classes | |||||||
ER | Negative | 140 (49.6) | 142 (50.4) | NS | 59 (18.9) | 253 (81.1) |
0.041
|
Positive | 405 (51.7) | 379 (48.3) | 134 (14.1) | 815 (85.9) | |||
HER2 | Negative | 469 (52.1) | 432 (47.9) | NS | 165 (15.3) | 913 (84.7) | NS |
Positive | 63 (46.3) | 73 (53.7) | 22 (14.0) | 135 (86.0) | |||
Triple negative | No | 446 (52.1) | 410 (47.9) | NS | 145 (14.1) | 883 (85.9) |
0.030
|
Yes | 90 (47.6) | 99 (52.4) | 42 (20.0) | 168 (80.0) | |||
Basal phenotype | No | 414 (52.0) | 382 (48.0) | NS | 141 (14.8) | 810 (85.2) | NS |
Yes | 138 (48.9) | 132 (51.1) | 53 (16.8) | 263 (83.2) |
Association with clinicopathological and molecular variables sporadic breast cancer
PARP1nc
Variables | PARP1 non-cleaved | PARP1 cleaved | ||||
---|---|---|---|---|---|---|
Negative n (%) | Positive n (%) |
p value | Negative n (%) | Positive n (%) |
p value | |
Menopausal status | ||||||
Pre | 182 (33.2) | 216 (41.7) |
0.004
| 65 (33.5) | 409 (38.5) | NS |
Post | 366 (66.8) | 302 (58.3) | 129 (66.5) | 653 (61.5) | ||
Age | ||||||
<50 | 163 (29.7) | 193 (36.9) |
0.012
| 61 (31.4) | 365 (34) | NS |
≥50 | 386 (70.3) | 330 (63.1) | 133 (68.6) | 710 (66) | ||
Tumour size | ||||||
≤2 cm | 279 (51.2) | 240 (46.1) | NS | 96 (50) | 523 (48.8) | NS |
>2 cm | 266 (48.8) | 281 (53.9) | 96 (50) | 548 (51.2) | ||
Tumour stage | ||||||
1 | 346 (62.9) | 310 (59.5) | NS | 129 (66.5) | 634 (59.2) | NS |
2 | 159 (28.9) | 164 (31.5) | 51 (26.3) | 342 (31.9) | ||
3 | 45 (8.2) | 47 (9.0) | 14 (7.2) | 95 (8.9) | ||
Tumour grade | ||||||
1 | 113 (20.7) | 69 (13.3) |
0.001
| 32 (16.7) | 180 (17.9) | NS |
2 | 180 (33) | 163 (31.3) | 61 (31.8) | 368 (34.5) | ||
3 | 253 (46.3) | 288 (55.4) | 99 (51.6) | 519 (48.6) | ||
Tubule formation | ||||||
1 | 35 (6.7) | 24 (4.7) | NS | 11 (5.9) | 62 (6) | NS |
2 | 181 (34.6) | 160 (31.6) | 57 (30.3) | 336 (32.6) | ||
3 | 307 (58.7) | 323 (63.7) | 120 (63.8) | 634 (61.4) | ||
Pleomorphism | ||||||
1 | 14 (2.7) | 10 (2) |
<0.001
| 1 (0.5) | 29 (2.8) | NS |
2 | 233 (44.6) | 168 (33.1) | 76 (40.4) | 404 (39.2) | ||
3 | 275 (52.7) | 329 (64.9) | 111 (59) | 598 (58) | ||
Mitosis | ||||||
1 | 201 (38.4) | 145 (28.6) |
0.001
| 66 (35.1) | 373 (36.1) | NS |
2 | 103 (19.7) | 95 (18.7) | 30 (16) | 200 (19.4) | ||
3 | 219 (41.9) | 267 (52.7) | 92 (48.9) | 459 (44.5) | ||
Vascular invasion | ||||||
Negative | 185 (34) | 174 (33.5) | NS | 65 (34) | 355 (33.2) | NS |
Positive | 359 (66) | 346 (66.5) | 126 (66) | 714 (66.8) | ||
NPI groups | ||||||
Good | 190 (34.9) | 134 (25.7) |
0.005
| 63 (32.8) | 326 (30.4) | NS |
Moderate | 269 (49.4) | 295 (56.6) | 94 (49) | 566 (52.8) | ||
Poor | 86 (15.8) | 92 (17.7) | 35 (18.2) | 179 (16.7) |
Variables | PARP1 non-cleaved | PARP1 cleaved | |||||
---|---|---|---|---|---|---|---|
Negative no. (%) | Positive no. (%) |
p value | Negative no. (%) | Positive no. (%) |
p value | ||
APE1 | Negative | 31 (16.2) | 36 (15.9) | NS | 16 (19.8) | 59 (14.4) | NS |
Positive | 160 (83.8) | 190 (84.1) | 65 (80.2) | 352 (85.6) | |||
BARD1 | Negative | 299 (87.4) | 314 (88.2) | NS | 115 (89.8) | 620 (86.8) | NS |
Positive | 43 (12.6) | 42 (11.8) | 13 (10.2) | 86 (12.2) | |||
BRCA1 | Negative | 218 (51.8) | 178 (41.9) |
0.004
| 74 (50.3) | 370 (43.3) | NS |
Positive | 203 (48.2) | 247 (58.1) | 73 (49.7) | 485 (56.7) | |||
BRCA2 | Negative | 348 (92.6) | 333 (92.5) | NS | 111 (92.5) | 666 (92.4) | NS |
Positive | 28 (7.4) | 27 (7.5) | 9 (7.5) | 55 (7.6) | |||
RAD51 | Negative | 275 (68.9) | 191 (48.1) |
<0.001
| 107 (84.3) | 391 (53.8) |
<0.001
|
Positive | 124 (31.1) | 206 (51.9) | 20 (15.7) | 336 (46.2) | |||
CHK1 | Negative | 360 (79.3) | 323 (69.3) |
<0.001
| 127 (84.7) | 607 (71.9) |
<0.001
|
Positive | 94 (20.7) | 143 (30.7) | 23 (15.3) | 237 (28.1) | |||
Ku70/Ku80 | Negative | 78 (17.3) | 65 (14.6) | NS | 61 (38.4) | 101 (12.5) |
<0.001
|
Positive | 373 (82.7) | 379 (85.4) | 98 (61.6) | 708 (87.5) | |||
PIASγ | Negative | 156 (33.3) | 74 (15.7) |
<0.001
| 80 (50) | 164 (18.3) |
<0.001
|
Positive | 113 (66.7) | 396 (84.3) | 80 (50) | 730 (81.7) | |||
DNA-PKcs | Negative | 81 (25.9) | 38 (11.3) |
<0.001
| 44 (36.7) | 88 (13.8) |
<0.001
|
Positive | 232 (74.1) | 299 (88.7) | 76 (63.3) | 552 (86.2) | |||
Ki67 | Negative | 145 (35.5) | 122 (28.2) |
0.022
| 49 (33.6) | 284 (33.1) | NS |
Positive | 263 (64.5) | 311 (71.8) | 97 (66.4) | 573 (66.9) | |||
ID4 | Negative | 424 (87.4) | 410 (89.5) | NS | 148 (85.1) | 839 (89.4) | NS |
Positive | 61 (12.6) | 48 (10.5) | 26 (14.9) | 99 (10.6) | |||
P53 | Negative | 413 (77.1) | 328 (65.3) |
<0.001
| 137 (75.3) | 748 (71.2) | NS |
Positive | 123 (22.9) | 174 (34.7) | 45 (24.7) | 302 (28.8) |
Marker | PARP1nc+/BRCA1−
n (%) | PARP1nc−/BRCA1+
n (%) |
p value | |
---|---|---|---|---|
CHK1 | Negative | 137 (52.3) | 125 (47.7) |
0.004
|
Positive | 21 (32.3) | 44 (67.7) | ||
DNA-PKcs | Negative | 12 (32.4) | 25 (67.6) |
0.003
|
Positive | 116 (59.2) | 80 (40.8) | ||
P53 | Negative | 99 (37.1) | 168 (62.9) |
<0.001
|
Positive | 78 (69.0) | 25 (31.0) | ||
PIASγ | Negative | 25 (36.2) | 44 (63.8) |
0.013
|
Positive | 143 (53.0) | 127 (47.0) | ||
Ki67 | Negative | 25 (29.8) | 59 (70.2) |
<0.001
|
Positive | 130 (61.3) | 82 (47.6) |
Variables | ER-positive | ER negative | |||||
---|---|---|---|---|---|---|---|
Negative n (%) | Positive n (%) |
p value | Negative n (%) | Positive n (%) |
p value | ||
PARP1 non-cleaved | |||||||
BRCA1 | Negative | 138 (44.5) | 105 (33.9) |
0.007
| 80 (72.1) | 73 (64) | NS |
Positive | 172 (55.5) | 205 (66.1) | 31 (27.9) | 41 (36) | |||
RAD51 | Negative | 197 (67) | 113 (39.9) |
<0.001
| 77 (75.5) | 78 (69.6) | NS |
Positive | 97 (33) | 170 (60.1) | 25 (24.5) | 34 (30.4) | |||
CHK1 | Negative | 256 (77.3) | 216 (64.1) |
<0.001
| 102 (85) | 106 (83.5) | NS |
Positive | 75 (22.7) | 121 (35.9) | 18 (15) | 21 (16.5) | |||
CHK2 | Negative | 135 (57.4) | 94 (39.8) |
<0.001
| 66 (64.1) | 54 (53.5) | NS |
Positive | 100 (42.6) | 142 (60.2) | 37 (35.9) | 47 (46.5) | |||
PIASγ | Negative | 111 (32.5) | 48 (14.3) |
<0.001
| 45 (36) | 26 (19.7) |
0.003
|
Positive | 231 (67.5) | 288 (85.7) | 80 (64) | 106 (80.3) | |||
DNA-PKcs | Negative | 58 (26) | 25 (10.5) |
<0.001
| 23 (25.8) | 13 (13.3) |
0.029
|
Positive | 165 (74) | 214 (89.5) | 66 (74.2) | 85 (86.7) | |||
Ki67 | Negative | 126 (42.3) | 114 (36.4) | NS | 18 (16.7) | 8 (6.7) |
0.019
|
Positive | 172 (57.7) | 199 (63.6) | 90 (83.3) | 111 (93.3) | |||
ID4 | Negative | 323 (91.2) | 302 (90.1) | NS | 98 (77.2) | 106 (87.6) |
0.031
|
Positive | 31 (8.8) | 33 (9.9) | 29 (22.8) | 15 (12.4) | |||
P53 | Negative | 339 (85.2) | 277 (75.7) |
0.001
| 73 (53.3) | 51 (37.5) |
0.009
|
Positive | 59 (14.8) | 89 (24.3) | 64 (46.7) | 85 (62.5) | |||
PARP1 cleaved | |||||||
RAD51 | Negative | 70 (81.4) | 264 (49.1) |
<0.001
| 37 (90.2) | 126 (68.9) |
0.010
|
Positive | 16 (18.6) | 274 (50.9) | 4 (9.8) | 57 (31.1) | |||
CHK1 | Negative | 85 (82.5) | 428 (67.9) |
0.003
| 42 (89.4) | 175 (84.1) | NS |
Positive | 18 (17.5) | 202 (32.1) | 5 (10.6) | 33 (15.9) | |||
CHK2 | Negative | 52 (66.7) | 214 (45.2) |
<0.001
| 27 (61.4) | 106 (58.9) | NS |
Positive | 26 (33.3) | 259 (54.8) | 17 (38.6) | 75 (41.1) | |||
PIASγ | Negative | 55 (50.9) | 113 (17) |
<0.001
| 25 (48.1) | 51 (22.9) |
<0.001
|
Positive | 53 (49.1) | 552 (83) | 27 (51.9) | 172 (77.1) | |||
DNA-PKcs | Negative | 26 (34.2) | 65 (13.6) |
<0.001
| 18 (40.9) | 23 (14.3) |
<0.001
|
Positive | 50 (65.8) | 413 (86.4) | 26 (59.1) | 138 (85.7) | |||
P53 | Negative | 112 (87.5) | 637 (79.8) |
0.040
| 25 (46.3) | 108 (43.4) | NS |
Positive | 16 (12.5) | 161 (20.2) | 29 (53.7) | 141 (56.6) |
Variables | PARP1 non-cleaved | PARP1 cleaved | |||||
---|---|---|---|---|---|---|---|
Negative n (%) | Positive n (%) |
p value | Negative n (%) | Positive n (%) |
p value | ||
PIASγ | Negative | 35 (42.7) | 19 (20.7) |
0.002
| 19 (51.4) | 38 (25) |
0.002
|
Positive | 47 (57.3) | 73 (79.3) | 18 (48.6) | 114 (75) | |||
DNA-PKcs | Negative | 15 (25.9) | 10 (15.2) | NS | 12 (41.4) | 16 (14.4) |
0.001
|
Positive | 43 (74.1) | 56 (84.8) | 17 (58.6) | 95 (85.6) | |||
P53 | Negative | 48 (54.5) | 37 (37.8) |
0.022
| 18 (45) | 71 (42.5) | NS |
Positive | 40 (45.5) | 61 (62.2) | 22 (55) | 96 (57.5) |