01.12.2017 | Research article | Ausgabe 1/2017 Open Access

Expression of enhancer of zeste homolog 2 correlates with survival outcome in patients with metastatic breast cancer: exploratory study using primary and paired metastatic lesions
- Zeitschrift:
- BMC Cancer > Ausgabe 1/2017
Electronic supplementary material
Background
Methods
Patients and samples
TMAs
IHC analysis
Biomarker
|
Primary lesions
|
Metastatic lesions
|
P-value
|
---|---|---|---|
ER status,
n (%)
|
|||
positive
|
51 (53.1)
|
41 (42.7)
|
0.149
|
negative
|
45 (46.9)
|
55 (57.3)
|
|
PR status,
n (%)
|
|||
positive
|
47 (49.0)
|
39 (40.6)
|
0.246
|
negative
|
49 (51.0)
|
57 (59.4)
|
|
HER2 status,
n (%)
|
|||
positive
|
16 (16.7)
|
14 (14.6)
|
0.691
|
negative
|
80 (83.3)
|
82 (85.4)
|
|
Ki-67 expression,
n (%)
|
|||
high
|
55 (57.3)
|
72 (75.0)
|
0.010
*
|
low
|
41 (42.7)
|
24 (25.0)
|
|
EZH2 expression,
n (%)
|
|||
high
|
54 (56.3)
|
79 (82.3)
|
<0.0001
*
|
low
|
42 (43.7)
|
17 (17.7)
|
|
Molecular subtype,
n (%)
|
|||
luminal A
a
|
31 (32.3)
|
19 (19.8)
|
0.304
|
luminal B
b
|
22 (22.9)
|
29 (30.2)
|
|
luminal HER2
c
|
5 (5.2)
|
5 (5.2)
|
|
HER2-type
d
|
11 (11.5)
|
9 (9.4)
|
|
TNBC
e
|
27 (28.1)
|
34 (35.4)
|
Follow-up
Statistical analyses
EZH2
|
r
|
P-value
|
|
---|---|---|---|
Primary ER status
|
Primary
|
−0.103
|
0.318
|
Primary PR status
|
Primary
|
−0.111
|
0.282
|
Primary HER2 status
|
Primary
|
0.361
|
<0.001
*
|
Primary Ki-67 expression
|
Primary
|
0.722
|
<0.0001
*
|
Metastatic ER status
|
Metastatic
|
−0.099
|
0.339
|
Metastatic PR status
|
Metastatic
|
−0.190
|
0.064
|
Metastatic HER2 status
|
Metastatic
|
0.306
|
0.002
*
|
Metastatic Ki-67 expression
|
Metastatic
|
0.685
|
<0.0001
*
|
Results
Clinicopathological characteristics of all patients with MBC in this study
Characteristic
|
Patients (
n = 96)
|
Percent
|
---|---|---|
Age, years (mean ± SD)
|
51 ± 7.5
|
|
Menopausal status
|
||
pre-
|
44
|
45.8
|
post-
|
52
|
54.2
|
Tumor size
|
||
≤ 20 mm
|
17
|
17.7
|
> 20 mm
|
77
|
80.2
|
unknown
|
2
|
2.1
|
LN status
|
||
positive
|
63
|
65.6
|
negative
|
33
|
34.4
|
Histological type
|
||
ductal
|
83
|
86.5
|
special
a
|
13
|
13.5
|
LVI status
|
||
positive
|
63
|
65.6
|
negative
|
25
|
26.1
|
unknown
|
8
|
8.3
|
Operation status
|
||
partial mastectomy
|
17
|
17.8
|
mastectomy
|
79
|
82.2
|
Stage at the primary diagnosis
|
||
1
|
11
|
11.5
|
2
|
48
|
50
|
3
|
34
|
35.4
|
4
|
1
|
1.0
|
unknown
|
2
|
2.0
|
Chemotherapy
|
||
adjuvant
|
83
|
86.5
|
none
|
13
|
13.5
|
Hormone therapy
|
||
adjuvant
|
50
|
52.1
|
none
|
46
|
47.9
|
Site of recurrence
|
||
brain
|
26
|
27.0
|
lung
|
8
|
8.3
|
liver
|
1
|
1.0
|
ovary
|
2
|
2.1
|
chest wall
|
13
|
13.5
|
lymph node
|
15
|
15.6
|
distant skin
|
7
|
7.3
|
bone
|
24
|
25
|
Changes in ER, PR, HER2, Ki-67 and EZH2 expression between primary and metastatic lesions
Correlation coefficients of ER, PR, HER2, Ki-67 and EZH2 expression scores in primary and metastatic lesions
Relationship between EZH2 expression and patient clinicopathological characteristics
Characteristic
|
Primary EZH2
|
P-value
|
Metastatic EZH2
|
P-value
|
||
---|---|---|---|---|---|---|
low (
n = 42)
|
high (
n = 54)
|
low (
n = 17)
|
high (
n = 79)
|
|||
Age, years (mean ± SD)
|
52 ± 8.5
|
50 ± 9.8
|
0.198
|
50 ± 7.8
|
51 ± 9.7
|
0.241
|
Menopausal status,
n (%)
|
||||||
pre-
|
17 (38.6)
|
27 (61.4)
|
0.353
|
10 (22.7)
|
34 (77.3)
|
0.236
|
post-
|
25 (48.1)
|
27 (51.9)
|
7 (13.5)
|
45 (86.5)
|
||
Tumor size,
n (%)
|
||||||
≤ 20 mm
|
8 (47.1)
|
9 (52.9)
|
2 (11.8)
|
15 (88.2)
|
||
> 20 mm
|
33 (42.9)
|
44 (57.1)
|
0.936
|
14 (18.2)
|
63 (81.8)
|
0.396
|
unknown
|
1 (50.0)
|
1 (50.0)
|
1 (50.0)
|
1 (50.0)
|
||
LN status,
n (%)
|
||||||
positive
|
23 (36.5)
|
40 (63.5)
|
11 (17.5)
|
52 (82.5)
|
||
negative
|
19 (57.6)
|
14 (42.4)
|
0.048
*
|
6 (18.2)
|
27 (81.8)
|
0.930
|
Histological type
, n (%)
|
||||||
ductal
|
32 (38.6)
|
51 (61.4)
|
0.010
*
|
12 (14.5)
|
71 (85.5)
|
0.035
*
|
special
|
10 (76.9)
|
3 (23.1)
|
5 (38.5)
|
8 (61.5)
|
||
LVI status,
n (%)
|
||||||
positive
|
32 (50.8)
|
31 (49.2)
|
11 (17.5)
|
52 (82.5)
|
||
negative
|
8 (32.0)
|
17 (68.0)
|
0.148
|
4 (16.0)
|
21 (84.0)
|
0.842
|
unknown
|
2 (25.0)
|
6 (75.0)
|
2 (25.0)
|
6 (75.0)
|
||
Operation status,
n (%)
|
||||||
partial mastectomy
|
4 (23.5)
|
13 (76.5)
|
0.064
|
2 (11.8)
|
15 (88.2)
|
0.479
|
mastectomy
|
38 (48.1)
|
41 (51.9)
|
15 (19.0)
|
64 (81.0)
|
||
Chemotherapy,
n (%)
|
||||||
adjuvant
|
34 (41.0)
|
49 (59.0)
|
0.164
|
12 (14.5)
|
71 (85.5)
|
0.035
*
|
none
|
8 (61.5)
|
5 (38.5)
|
5 (38.5)
|
8 (61.5)
|
||
Hormone therapy,
n (%)
|
||||||
adjuvant
|
26 (52.0)
|
24 (48.0)
|
0.089
|
12 (24.0)
|
38 (76.0)
|
0.092
|
none
|
16 (34.8)
|
30 (65.2)
|
5 (10.9)
|
41 (89.1)
|
||
Primary ER status,
n (%)
|
||||||
positive
|
28
|
23
|
0.019
*
|
12
|
39
|
0.112
|
negative
|
14
|
31
|
5
|
40
|
||
Primary PR status,
n (%)
|
||||||
positive
|
24
|
23
|
0.157
|
12
|
35
|
0.049
*
|
negative
|
18
|
31
|
5
|
44
|
||
Primary HER2 status,
n (%)
|
||||||
positive
|
1
|
15
|
0.001
*
|
0
|
16
|
0.042
*
|
negative
|
41
|
39
|
17
|
63
|
||
Primary Ki-67 expression,
n (%)
|
||||||
high
|
8
|
47
|
<0.001
*
|
2
|
53
|
<0.001
*
|
low
|
34
|
7
|
15
|
26
|
||
Metastatic ER status,
n (%)
|
||||||
positive
|
21
|
20
|
0.203
|
8
|
33
|
0.689
|
negative
|
21
|
34
|
9
|
46
|
||
Metastatic PR status,
n (%)
|
||||||
positive
|
22
|
17
|
0.039
*
|
9
|
30
|
0.254
|
negative
|
20
|
37
|
8
|
49
|
||
Metastatic HER2 status,
n (%)
|
||||||
positive
|
1
|
13
|
0.003
*
|
0
|
14
|
0.060
|
negative
|
41
|
41
|
17
|
65
|
||
Metastatic Ki-67 expression,
n (%)
|
||||||
high
|
23
|
49
|
<0.001
*
|
2
|
70
|
<0.001
*
|
low
|
19
|
5
|
15
|
9
|
||
Site of recurrence,
n (%)
|
||||||
viscera
b
|
14 (37.8)
|
23 (62.2)
|
0.102
|
5 (13.5)
|
32 (86.5)
|
0.012
*
|
soft tissue
c
|
13 (37.1)
|
22 (62.9)
|
3 (8.6)
|
32 (91.4)
|
||
bone
|
15 (62.5)
|
9 (37.5)
|
9 (37.5)
|
15 (62.5)
|
||
Disease free interval
|
||||||
≤ 2 years
|
7 (23.5)
|
24(77.4)
|
0.04
*
|
4 (12.9)
|
27 (87.1)
|
0.394
|
> 2 years
|
35 (53.8)
|
30 (46.1)
|
13 (20)
|
52 (80)
|
||
≤ 10 years
|
32 (38.6)
|
51 (61.4)
|
0.010
*
|
13 (15.7)
|
70 (84.3)
|
0.185
|
> 10 years
|
10 (76.9)
|
3 (23.1)
|
4 (30.8)
|
9 (69.2)
|
Relationship between EZH2 expression and patient outcome
Prognostic factor
|
Patients (
n = 96)
|
Univariate analysis
|
Multivariate analysis
|
||||
---|---|---|---|---|---|---|---|
HR
|
95% CI
|
P-value
|
HR
|
95% CI
|
P-value
|
||
Menopausal status,
n (%)
|
|||||||
pre-
|
44 (45.8)
|
1
|
|||||
post-
|
52 (54.2)
|
1.396
|
0.891–2.186
|
0.145
|
|||
Tumor size,
n (%)
|
|||||||
≤ 20 mm
|
17 (17.7)
|
1
|
|||||
> 20 mm
|
77 (80.2)
|
1.317
|
0.726–2.388
|
0.364
|
|||
unknown
|
2 (2.1)
|
||||||
LN status,
n (%)
|
|||||||
positive
|
63 (65.6)
|
1.309
|
0.829–2.066
|
0.248
|
|||
negative
|
33 (34.4)
|
1
|
|||||
Histological type,
n (%)
|
|||||||
ductal
|
83 (86.5)
|
1
|
|||||
special
a
|
13 (13.5)
|
1.028
|
0.543–1.945
|
0.932
|
|||
LVI status,
n (%)
|
|||||||
positive
|
63 (65.6)
|
1.287
|
0.769–2.156
|
0.337
|
|||
negative
|
25 (26.1)
|
1
|
|||||
unknown
|
8 (8.3)
|
||||||
Operation status,
n (%)
|
|||||||
partial mastectomy
|
17 (17.7)
|
1
|
|||||
mastectomy
|
79 (82.3)
|
0.894
|
0.500–1.596
|
0.704
|
|||
Chemotherapy,
n (%)
|
|||||||
adjuvant
|
83 (86.5)
|
1.320
|
0.707–2.461
|
0.383
|
|||
none
|
13 (13.5)
|
1
|
|||||
Hormone therapy,
n (%)
|
|||||||
adjuvant
|
50 (52.1)
|
0.719
|
0.460–1.122
|
0.146
|
|||
none
|
46 (47.9)
|
1
|
|||||
Primary ER status,
n (%)
|
|||||||
positive
|
51 (53.1)
|
0.541
|
0.344–0.850
|
0.008
*
|
0.724
|
0.382–1.372
|
0.322
|
negative
|
45 (46.9)
|
1
|
|||||
Primary PR status,
n (%)
|
|||||||
positive
|
47 (49.0)
|
0.595
|
0.380–0.931
|
0.023
*
|
0.911
|
0.484–1.714
|
0.773
|
negative
|
49 (51.0)
|
1
|
|||||
Primary HER2 status,
n (%)
|
|||||||
positive
|
16 (16.7)
|
1.122
|
0.616–2.046
|
0.706
|
|||
negative
|
80 (83.3)
|
1
|
|||||
Primary Ki-67 expression,
n (%)
b
|
|||||||
high
|
55 (57.3)
|
1.848
|
1.170–2.917
|
0.008
*
|
|||
low
|
41 (42.7)
|
1
|
|||||
Primary EZH2 expression,
n (%)
|
|||||||
high
|
54 (56.3)
|
1.449
|
0.930–2.258
|
0.101
|
|||
low
|
42 (43.7)
|
1
|
|||||
Metastatic ER status,
n (%)
|
|||||||
positive
|
41 (42.7)
|
0.972
|
0.625–1.513
|
0.901
|
|||
negative
|
55 (57.3)
|
1
|
|||||
Metastatic PR status,
n (%)
|
|||||||
positive
|
39 (40.6)
|
0.692
|
0.440–1.089
|
0.112
|
|||
negative
|
57 (59.4)
|
1
|
|||||
Metastatic HER2 status,
n (%)
|
|||||||
positive
|
14 (14.6)
|
1.282
|
0.673–2.440
|
0.450
|
|||
negative
|
82 (85.4)
|
1
|
|||||
Metastatic Ki-67 expression,
n (%)
b
|
|||||||
high
|
72 (75.0)
|
2.422
|
1.394–4.206
|
0.002
*
|
|||
low
|
24 (25.0)
|
1
|
|||||
Metastatic EZH2 expression,
n (%)
|
|||||||
high
|
59 (77.6)
|
2.116
|
1.143–3.916
|
0.017
*
|
2.047
|
1.074–3.902
|
0.029
*
|
low
|
17 (22.4)
|
1
|
|||||
Metastatic sites, n (%)
|
|||||||
bones
|
24 (25.0)
|
0.899
|
0.548–1.476
|
0.674
|
|||
others
|
72 (75.0)
|
1
|
|||||
Disease free interval
|
|||||||
≤ 2 years
|
28 (29.0)
|
1.713
|
1.086–2.702
|
0.021
*
|
1.601
|
0.978–2.621
|
0.061
|
> 2 years
|
68 (71.0)
|
1
|
|||||
≤ 10 years
|
83 (87.0)
|
1
|
|||||
> 10 years
|
13 (13.0)
|
0.861
|
0.442–1.679
|
0.661
|