Fig. 1Epidermal growth factor receptor protein expression in invasive ductal carcinoma: (A) 0, (B) 1+, (C) 2+, and (D) 3+.
Fig. 2Change in epidermal growth factor receptor gene copy number: (A) amplification, (B) high polysomy, (C) low trisomy, and (D) disomy.
Fig. 3Group 1 with HER-2+ (A) and EGFR+ (B), group 2 with EGFR+ (C) and CK5/6- (D), and group 3 with EGFR+ (E) and CK5/6+ (F). HER-2, human epidermal growth factor receptor; EGFR, epidermal growth factor receptor; CK, cytokeratin.
Fig. 4Kaplan-Meier disease-free survival curve. Patients in group 2 have better disease-free survival than in patients in group 1 and 3.
Table 1.Characteristics of the primary antibodies
Antibody |
Type |
Clone |
Source |
Dilution |
ER |
Monoclonal mouse |
1D5 |
Dako, Glostrup, Denmark |
1:50 |
PR |
Monoclonal mouse |
PgR 636 |
Dako, Glostrup, Denmark |
1:50 |
HER-2 |
Monoclonal rabbit |
SP3 |
Thermo Scientific, Rockford, IL, USA |
1:100 |
CK5/6 |
Monoclonal mouse |
D5/16B4 |
Dako, Glostrup, Denmark |
1:50 |
EGFR |
Monoclonal mouse |
E30 |
Dako, Glostrup, Denmark |
1:100 |
p53 |
Monoclonal mouse |
DO-7 |
Novocastra, Newcast upon Tyne, UK |
1:50 |
Ki-67 |
Monoclonal mouse |
MIB-1 |
Dako, Glostrup, Denmark |
1:50 |
Table 2.Immunohistochemistry scoring of EGFR protein expression according to HercepTest criteria
Grade |
Criteria |
0 |
No staining or membranous staining in < 10% of the tumor cells |
1+ |
Incomplete membrane staining in > 10% of the tumor cells |
2+ |
Weak to moderate complete membrane staining in > 10% of the tumor cells |
3+ |
Strong and complete membrane staining in > 10% of the tumor cells |
Table 3.
EGFR gene copy number changes according to group
|
|
Group 1 (HER-2+, CK5/6–, EFGR+) |
Group 2 (HER-2–, CK5/6–, EGFR+) |
Group 3 (HER-2–, CK5/6+, EGFR+) |
Total |
FISH+ |
Amplification |
7 (41.2) |
1 (5.6) |
8 (17.0) |
16 (19.5) |
High polysomy |
0 (0.0) |
1 (5.6) |
1 (2.1) |
2 (2.5) |
Subtotal |
7 (41.2) |
2 (11.2) |
9 (19.1) |
18 (22.0) |
FISH– |
Disomy |
8 (47.0) |
10 (55.6) |
10 (21.3) |
28 (34.1) |
Low trisomy |
2 (11.8) |
1 (5.6) |
14 (29.8) |
17 (20.7) |
High trisomy |
0 (0.0) |
1 (5.6) |
2 (4.3) |
3 (3.7) |
Low polysomy |
0 (0.0) |
4 (22.0) |
12 (25.5) |
16 (19.5) |
Subtotal |
10 (58.8) |
16 (88.8) |
38 (80.9) |
64 (78.0) |
Total |
|
17 |
18 |
47 |
82 |
Table 4.Correlation between EGFR protein expression and gene copy number changes
|
|
Protein 1+ |
Protein 2+ |
Protein 3+ |
Total |
FISH+ |
Amplification |
1 (8.3) |
6 (13.6) |
9 (34.6) |
16 (19.5) |
High polysomy |
0 (0.0) |
1 (2.3) |
1 (3.9) |
2 (2.5) |
Subtotal |
1 (8.3) |
7 (15.9) |
10 (38.5) |
18 (22.0) |
FISH– |
Disomy |
3 (25.0) |
16(36.3) |
9 (34.6) |
28 (34.1) |
Low trisomy |
5 (41.7) |
9 (20.5) |
3 (11.5) |
17 (20.7) |
High trisomy |
0 (0.0) |
3 (6.8) |
0 (0.0) |
3 (3.7) |
Low polysomy |
3 (25) |
9 (20.5) |
4 (15.4) |
16 (19.5) |
Subtotal |
11 (91.7) |
37 (84.1) |
16 (61.5) |
64 (78.0) |
Total |
|
12 |
44 |
26 |
82 |
Table 5.Comparison of the clinicopathological characteristics of the patients among the groups
Parameters |
Group 1 (HER-2+, CK5/6–, EFGR+) |
Group 2 (HER-2–, CK5/6–, EGFR+) |
Group 3 (HER-2–, CK5/6+, EGFR+) |
Total |
p-value |
Age (yr) |
|
|
|
|
|
≤ 50 |
6 (35) |
9 (50) |
26 (55) |
41 (50) |
G1 vs G2: 0.380 |
> 50 |
11 (65) |
9 (50) |
21 (45) |
41 (50) |
G2 vs G3: 0.367 |
Mean age (yr) |
53.0 |
52.4 |
48.1 |
50.4 |
G1 vs G3: 0.157 |
Tumor size (cm) |
|
|
|
|
|
≤ 2 |
14 (82) |
7 (39) |
26 (55) |
47 (57) |
G1 vs G2: 0.009 |
> 2 |
3 (18) |
11 (61) |
21 (45) |
35 (43) |
G2 vs G3: 0.031 |
|
|
|
|
|
G1 vs G3: 0.048 |
Histologic grade |
|
|
|
|
|
1 |
1 (6) |
1 (6) |
0 (0) |
2 (2) |
G1 vs G2: 0.694 |
2 |
8 (47) |
6 (33) |
3 (6) |
17 (21) |
G2 vs G3: 0.001 |
3 |
8 (47) |
11 (61) |
44 (94) |
63 (77) |
G1 vs G3: 0.000 |
LN metastasis |
|
|
|
|
|
Negative |
11 (65) |
16 (89) |
37 (79) |
64 (78) |
G1 vs G2: 0.089 |
Positive |
6 (35) |
2 (11) |
10 (21) |
18 (22) |
G2 vs G3: 0.222 |
|
|
|
|
|
G1 vs G3: 0.253 |
Ki-67 (%) |
|
|
|
|
|
≤ 60 |
13 (76) |
5 (28) |
19 (40) |
37 (45) |
G1 vs G2: 0.004 |
> 60 |
4 (24) |
13 (72) |
28 (60) |
45 (55) |
G2 vs G3: 0.009 |
|
|
|
|
|
G1 vs G3: 0.011 |
p53 (%) |
|
|
|
|
|
≤ 60 |
13 (76) |
11 (61) |
26 (55) |
50 (61) |
G1 vs G2: 0.328 |
> 60 |
4 (24) |
7 (39) |
21 (45) |
32 (39) |
G2 vs G3: 0.309 |
|
|
|
|
|
G1 vs G3: 0.126 |
Table 6.Summary of the treatment modality in addition to surgery
Treatment |
Group 1 (HER-2+, CK5/6–, EFGR+) |
Group 2 (HER-2–, CK5/6–, EGFR+) |
Group 3 (HER-2–, CK5/6+, EGFR+) |
Chemotherapy |
5 (1) |
4 (1)a
|
17 (3) |
Herceptin therapy |
|
|
1 (1) |
Radiotherapy |
1 |
2 |
4 (2) |
Chemotherapy and Herceptin therapy |
2 (1) |
|
|
Chemotherapy and radiotherapy |
1 |
9 (1) |
23 |
Herceptin therapy and radiotherapy |
3 (2) |
|
|
Radiotherapy and hormone therapy |
|
|
1 (1) |
Chemotherapy, Herceptin therapy and radiotherapy |
3 |
|
|
None |
2 (1) |
3 |
1 |
Total |
17 (5) |
18 (2)a
|
47 (7) |