Background
Methods
Patient samples
Feature | ||
---|---|---|
Age (years) | Median = 62.5 | Range: 30–85 |
Mutation carrier status | ||
BRCA1
| 3 | 5.0% |
BRCA2
| 25 | 41.7% |
BRCAX
| 32 | 53.3% |
Size (mm) | Median = 17 | Range: 2–50 |
Histological subtype | ||
Invasive carcinoma - no special type (IC-NST) | 46 | 76.7% |
Invasive papillary carcinoma | 8 | 13.3% |
IC-NST with areas of micropapillary | 4 | 6.7% |
Invasive lobular carcinoma | 2 | 3.3% |
Grade | ||
1 | 2 | 3.3% |
2 | 30 | 50.0% |
3 | 28 | 46.7% |
DCIS | ||
Present | 41 | 68.3% |
Absent | 15 | 25.0% |
Unknown | 4 | 6.7% |
Nodal Status | ||
N0 | 28 | 46.7% |
N1 | 20 | 33.3% |
Nx | 12 | 20.0% |
Paget’s Disease | ||
Present | 8 | 13.3% |
Absent | 44 | 73.3% |
Unknown | 8 | 13.3% |
ER status (Allred score) | ||
Negative (0–4/8) | 2 | 3.3% |
Positive (5–8/8) | 58 | 96.7% |
PgR status (allred score) | ||
Negative (0–4/8) | 8 | 13.3% |
Positive (5–8/8) | 52 | 86.7% |
HER2 (SISH) | ||
Amplified | 5 | 8.3% |
Non-amplified | 55 | 91.7% |
Phenotype | ||
Luminal | 54 | 90.0% |
HER2 | 5 | 8.3% |
Basal | 1 | 1.7% |
Germline BRCA1/2 testing
DNA extraction
Bisulfite modification
Methylation-sensitive high resolution melting (MS-HRM)
Methylation scoring
Cluster analysis
Statistical analysis
Results
Methylation analysis of MBCs finds associations with genotype and clinico-pathological characteristics
GSTP1
|
RASSF1A
|
MAL
|
TWIST
|
RUNX3
|
RARβ
|
APC
|
FOXC1
|
CDH1
|
WIF1
| TOTAL HYPERMETHYLATED GENES | AMI (mean) | |
---|---|---|---|---|---|---|---|---|---|---|---|---|
BRCA1 (n = 3) | 2 (66%) | 1 (33%) | 1 (33%) | 0 | 1 (33%) | 1 (33%) | 1 (33%) | 1 (33%) | 0 | 1 (33%) | 9 (30%) | 13.4 |
BRCA2 (n = 25) | 22 (88%) | 20 (80%) | 14 (56%) | 13 (52%) | 8 (32%) | 11 (44%) | 8 (32%) | 6 (24%) | 0 | 11 (44%) | 113 (45%) | 23.6 |
BRCAX (n = 32) | 25 (78%) | 20 (63%) | 12 (38%) | 9 (28%) | 9 (28%) | 6 (19%) | 7 (22%) | 8 (25%) | 0 | 14 (56%) | 110 (34%) | 17.0 |
All (n = 60) | 49 (82%) | 41 (68%) | 27 (45%) | 22 (37%) | 18 (30%) | 18 (30%) | 16 (27%) | 15 (25%) | 0 | 26 (43%) | 232 (39%) | 14.0 |
p = 0.06 |
p = 0.08 |
P < 0.01 |
p = 0.01 |
GSTP1
|
RASSF1A
|
MAL
|
RUNX3
|
RARβ
|
APC
|
FOXC1
| AMI (median) | |||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Hypermethylation | + | - | + | - | + | - | + | - | + | - | + | - | + | - | > | < |
Age (years) | 59.1 | 65.7 | 67.2 | 60.9 | 69.1 | 60.4 | ||||||||||
p-value |
0.04
| 0.07 |
0.01
| |||||||||||||
Tumour size (mm) | 22.3 | 16.5 | 21.4 | 17.1 | 20.8 | 15.8 | ||||||||||
p-value |
0.01
| 0.08 |
0.02
| |||||||||||||
IC-NST Histology | 94% | 69% | ||||||||||||||
p-value |
0.046
| |||||||||||||||
Grade 3 | 51% | 18% | ||||||||||||||
p-value | 0.09 | |||||||||||||||
Paget’s Disease | 31% | 8% | ||||||||||||||
p-value |
0.04
| |||||||||||||||
DCIS present | 33% | 6% | ||||||||||||||
p-value |
0.02
| |||||||||||||||
Lymphovascular invasion | 49% | 18% | 20% | 51% | 53% | 40% | ||||||||||
p-value | 0.09 | 0.07 | 0.09 | |||||||||||||
Perineural invasion | 63% | 36% | ||||||||||||||
p-value | 0.07 | |||||||||||||||
Node positive | 52% | 24% | 62% | 34% | ||||||||||||
p-value | 0.08 | 0.07 | ||||||||||||||
HER2 positive | 13% | 0 | ||||||||||||||
p-value | 0.11 |