Background
Methods
Patients and histological samples
Tissue microarray construction
Fluorescence In Situ Hybridization on TMA analysis
Immunohistochemistry analysis
Protein | Antibody | Origin | Clone | Dilution |
---|---|---|---|---|
Estrogen receptor | mmab | Novocastra Laboratories | 6F11 | 1/60 |
Progesterone receptor | mmab | DakoCytomation | PgR 636 | 1/80 |
ERBB2 | rpab | DakoCytomation | HercepTest | 1/400 |
P53 | mmab | Immunotech | DO-1 | 1/4 |
Ki67 | mmab | DakoCytomation | MIB-1 | 1/100 |
Statistical methods
Results
Frequencies of amplifications and co-amplifications
Region of amplification | Gene regions | Number of FISH informative cases | % of tumors with amplification |
---|---|---|---|
8p12 |
RAB11FIP1
| 279 | 13.3% |
FGFR1
| 319 | 9.4% | |
20q13 |
MYBL2
| 265 | 5.7% |
ZNF217
| 233 | 9.9% | |
AURKA
| 282 | 3.5% |
Correlation of amplified regions with histoclinical factors
Correlation of amplified regions with clinical outcome
Region of amplification | Population | 5-year M FS [95% CI] | Hazard ratio [95% CI] |
p-value | |||
---|---|---|---|---|---|---|---|
W hole | N + | N - | With amplification | Without amplification | |||
8p12 | + | 68.75 [56.73–83.3] | 82.4 [76.6–88.7] | 2.25 [1.28–3.93] |
0.0046
| ||
8p12 | + | 63.2 [46.6–85.6] | 72.6 [63–83.67] | 1.87 [0.95–3.71] | 0.071 | ||
8p12 | + | 73.77 [57.59–94.49] | 91.33 [85.37–97.70] | 2.98 [1.1–8.06] |
0.031
| ||
8q24 | + | 78.6 [59.8–100] | 82.61 [77.63–87.92] | 0.825 [0.25–2.75] | 0.75 | ||
8q24 | + | 75 [50.3–100] | 74.53 [66.32–83.75] | 0.728 [0.17–3.04] | 0.66 | ||
8q24 | + | 83.3 [58.3–100] | 89.55 [84.09–95.36] | 0.825 [0.09–7.66] | 0.87 | ||
11q13 | + | 75.93 [65.28–88.31] | 83.95 [79.18–89] | 1.46 [0.8–2.67] | 0.22 | ||
11q13 | + | 75.72 [61.54–93.17] | 74 [65.7–83.4] | 0.962 [0.44–2.1] | 0.92 | ||
11q13 | + | 76.02 [60.88–94.92] | 91.72 [86.91–96.80] | 2.46 [0.93–6.48] | 0.069 | ||
12p13 | + | 91.67 [77.29–100] | 82.23 [77.85–86.85] | 0.427 [0.06–3.08] | 0.4 | ||
12p13 | + | 100 [100-100] | 72.14 [64.84–80.26] | * | 1 | ||
12p13 | + | 83.3 [58.3–100] | 91.28 [86.85–95.94] | 2.46 [0.32–18.75] | 0.39 | ||
17q12 | + | 64.7 [47.5–88.2] | 83.92 [79.28–88.83] | 2.09 [1.02–4.26] |
0.044
| ||
17q12 | + | 68.2 [48.6–95.7] | 75.37 [67.54–84.10] | 1.24 [0.52–2.95] | 0.63 | ||
17q12 | + | 55.6 [27.4–100] | 91.25 [86.44–96.33] | 4.2 [1.18–14.95] |
0.027
| ||
20q13Z
| + | 85.4 [71.27–100] | 79.43 [73.82–85.47] | 0.57 [0.18–1.83] | 0.35 | ||
20q13Z
| + | 90.91 [75.41–100] | 63.98 [54.65–74.89] | 0.186 [0.03–1.35] | 0.097 | ||
20q13Z
| + | 79.5 [57.7–100] | 93.05 [88.18–98.19] | 2.58 [0.55–12.15] | 0.23 | ||
20q13Co
| + | 90.43 [78.63–100] | 79.80 [74.71–85.24] | 0.364 [0.09–1.51] | 0.16 | ||
20q13Co
| + | 87.84 [73.37–100] | 66.80 [58.20–76.66] | 0.276 [0.07–1.15] | 0.076 | ||
20q13Co
| + | 100 [100-100] | 90.40 [85.35–95.75] | * | 1 |
Variable | Coefficient value | Hazard ratio 95% CI |
p-value |
---|---|---|---|
8p12 | 0.925 | 0.15 | |
Non Amplified | 1 | ||
Amplified | 2.52 (0.72–8.79) | ||
Ki67 | 1.29 | 0.055 | |
<20 | 1 | ||
≥20 | 3.63 (0.97–13.52) |
Correlation of amplified regions with molecular subtypes
Region of amplification | Basal | Non Basal |
p-value | ERBB2 | Non ERBB2 |
p-value | Luminal A | Non Luminal A |
p-value |
---|---|---|---|---|---|---|---|---|---|
No. of patients (%) | No. of patients (%) | No. of patients (%) | |||||||
11q13 | |||||||||
Amplification
| 1 (5.5) | 22 (45.8) |
p
= 0.006
| 2 (25) | 21 (36.2) | NS | 19 (57.6) | 4 (12.1) |
p
= 0.0003
|
No Amplification
| 17 (94.5) | 26 (54.2) | 6 (75) | 37 (63.8) | 14 (42.4) | 29 (87.9) | |||
12p13 | |||||||||
Amplification
| 5 (27.8) | 2 (4.2) |
p
= 0.02
| 1 (11.1) | 6 (10.5) | NS | 1 (3.1) | 6 (17.6) | NS |
No Amplification
| 13 (72.2) | 46 (95.8) | 8 (88.9) | 51 (89.5) | 31 (96.9) | 28 (82.4) | |||
17q12 | |||||||||
Amplification
| 0 (0) | 6 (17.6) | NS | 5 (71.4) | 1 (2.2) |
p
= 2.061e-06
| 0 (0) | 6 (20) | NS |
No Amplification
| 19 (100) | 28 (82.4) | 2 (28.6) | 45 (97.8) | 23 (100) | 24 (80) | p = 0.065 |