Background
Materials and methods
Patients
Variable | Total (n = 524) | FKBP12 high, no. (%) | FKBP12 low, no. (%) | P value | MDM2 high, no. (%) | MDM2 low, sno. (%) | P value |
---|---|---|---|---|---|---|---|
Age | |||||||
< 56 | 313 | 85 (27.2) | 228 (72.8) | 0.748 | 189 (60.4) | 124 (39.6) | 0.471 |
≥ 56 | 211 | 60 (28.4) | 151 (71.6) | 134 (63.5) | 77 (36.5) | ||
Menopausal status | |||||||
Premenopausal | 265 | 81 (30.6) | 184 (69.4) | 0.134 | 156 (58.9) | 109 (41.1) | 0.187 |
Postmenopausal | 259 | 64 (24.7) | 195 (75.3) | 167 (64.5) | 92 (35.5) | ||
Size-TNM | |||||||
pT1 | 188 | 59 (31.4) | 129 (68.6) | 0.109 | 106 (56.4) | 82 (43.6) | 0.112 |
pT2 + pTx | 252 | 70 (27.8) | 182 (72.2) | 159 (63.1) | 93 (36.9) | ||
pT3 + pT4 | 84 | 16 (19.0) | 68 (81.0) | 58 (69.0) | 26 (31.0) | ||
Histologic grade | |||||||
Grade I | 97 | 33 (34.0) | 64 (66.0) | 0.264 | 48 (49.5) | 49 (50.5) | 0.015 |
Grade II | 217 | 61 (28.1) | 156 (71.9) | 135 (62.2) | 82 (37.8) | ||
Grade III | 210 | 51 (24.3) | 159 (75.7) | 140 (66.7) | 70 (33.3) | ||
LN metastasis | |||||||
Positive | 319 | 83 (26.0) | 236 (74.0) | 0.291 | 204 (63.9) | 115 (36.1) | 0.175 |
Negative | 205 | 62 (30.2) | 143 (69.8) | 119 (58.0) | 86 (42.0) | ||
NPI
| |||||||
< 3.4 | 180 | 57 (31.7) | 123 (68.3) | 0.107 | 84 (46.7) | 96 (53.3) | < 0.001 |
3.4–5.4 | 269 | 74 (27.5) | 195 (72.5) | 189 (70.3) | 80 (29.7) | ||
> 5.4 | 75 | 14 (18.7) | 61 (81.3) | 50 (66.7) | 25 (33.3) | ||
MIB-1 (%) | |||||||
< 10 | 204 | 63 (30.9) | 141 (69.1) | 0.134 | 117 (57.4) | 87 (42.6) | 0.028 |
10–30 | 254 | 70 (27.6) | 184 (72.4) | 171 (67.3) | 83 (32.7) | ||
> 30 | 66 | 12 (18.2) | 54 (81.8) | 35 (53.0) | 31 (47.0) | ||
ERα | |||||||
Positive | 343 | 107 (31.2) | 236 (68.8) | 0.013 | 200 (58.3) | 143 (41.7) | 0.031 |
Negative | 181 | 38 (21.0) | 143 (79.0) | 123 (68.0) | 58 (32.0) | ||
PR | |||||||
Positive | 319 | 98 (30.7) | 221 (69.3) | 0.052 | 187 (58.6) | 132 (41.4) | 0.076 |
Negative | 205 | 47 (22.9) | 158 (77.1) | 136 (66.3) | 69 (33.7) | ||
HER2 amplification | |||||||
Positive | 103 | 21 (20.4) | 82 (79.6) | 0.065 | 71 (68.9) | 32 (31.1) | 0.090 |
Negative | 421 | 124 (29.5) | 297 (70.5) | 252 (59.9) | 169 (40.1) | ||
Hormone therapy | |||||||
No | 182 | 41 (22.5) | 141 (77.5) | 0.120 | 121 (66.5) | 61 (33.5) | 0.114 |
Yes (Tamoxifen) | 332 | 102 (30.7) | 230 (69.3) | 198 (59.6) | 134 (40.4) | ||
Unknown | 10 | 2 (20.0) | 8 (80.0) | 4 (40.0) | 6 (60.0) | ||
Chemotherapy | |||||||
No | 31 | 9 (29.0) | 22 (71.0) | 0.235 | 19 (61.3) | 12 (38.7) | 0.118 |
Yes (FEC) | 469 | 133 (28.4) | 336 (71.6) | 294 (62.7) | 175 (37.3) | ||
Unknown | 24 | 3 (12.5) | 21 (87.5) | 10 (41.7) | 14 (58.3) | ||
Recurrence | |||||||
No | 387 | 115 (29.7) | 272 (70.3) | 0.079 | 228 (58.9) | 159 (41.1) | 0.031 |
Yes | 137 | 30 (21.9) | 107 (78.1) | 95 (69.3) | 42 (30.7) | ||
MDM2 expression | |||||||
High | 323 | 81 (25.1) | 242 (74.9) | 0.092 | |||
Low | 201 | 64 (31.8) | 137 (68.2) | ||||
FKBP12 expression | |||||||
High | 145 | 81 (55.9) | 64 (44.1) | 0.092 | |||
Low | 379 | 242 (63.9) | 137 (36.1) | ||||
Molecular subtype | |||||||
Luminal (non HER2+) | 334 | 102 (30.5) | 232 (69.5) | 0.144 | 196 (58.7) | 138 (41.3) | 0.171 |
HER2+ | 102 | 24 (23.5) | 78 (76.5) | 67 (65.7) | 35 (34.3) | ||
TNBC | 88 | 19 (21.6) | 69 (78.4) | 60 (68.2) | 28 (31.8) |
Variable | TNBC (n = 88) | FKBP12 high, no. (%) | FKBP12 low, no. (%) | P value | MDM2 high, no. (%) | MDM2 low, sno. (%) | P value |
---|---|---|---|---|---|---|---|
MDM2 high | 60 | 7 (11.7) | 53 (88.3) | 0.001 | |||
MDM2 low | 28 | 12 (42.9) | 16 (57.1) | ||||
FKBP12 high | 19 | 7 (36.8) | 12 (63.2) | 0.001 | |||
FKBP12 low | 69 | 53 (76.8) | 16 (23.2) |
Immunohistochemistry
Cell culture, plasmid transfection, and doxorubicin treatment
Western blot
WST-1 assay
Flow cytometry
Statistical analysis
Results
FKBP12 and MDM2 protein expression pattern in breast cancer tissue
FKBP12 and MDM2 expression correlated with clinicopathologic parameters
FKBP12 loss correlated with MDM2 overexpression
Correlation of FKBP12 expression with clinical outcome
Variable | Disease-free survival | Overall survival | ||||
---|---|---|---|---|---|---|
Hazard ratio | 95% CI | P value | Hazard ratio | 95% CI | P value | |
Age (< 56 vs ≥ 56) | 0.834 | 0.433–1.748 | 0.776 | 0.903 | 0.389–2.124 | 0.786 |
Menopausal status (pre. vs post.) | 0.687 | 0.325–1.975 | 0.477 | 0.525 | 0.223–1.867 | 0.335 |
Size (pT1-2 vs pT3-4) | 0.659 | 0.236–1.682 | 0.065 | 0.387 | 0.233–1.407 | 0.059 |
Histological grade (I–II vs III) | 0.412 | 0.237–1.531 | 0.158 | 0.704 | 0.498–1.994 | 0.086 |
LN metastasis (Posi. vs Neg.) | 0.627 | 0.247–1.033 | 0.014 | 0.754 | 0.162–1.228 | 0.019 |
NPI (< 3.4 vs ≥ 3.4) | 1.632 | 1.114–3.152 | 0.113 | 0.603 | 0.327–1.178 | 0.076 |
MIB (< 10% vs ≥ 10%) | 0.631 | 0.443–0.987 | 0.069 | 0.836 | 0.530–1.625 | 0.097 |
FKBP12 (high vs low) | 0.512 | 0.201–0.739 | < 0.001 | 0.317 | 0.186–0.586 | < 0.001 |
MDM2 (high vs low) | 0.405 | 0.427–0.855 | 0.003 | 0.372 | 0.264–0.679 | 0.005 |
Response to chemotherapy according to FKBP12 expression
Subtypes |
n
| PCR rate (FKBP12 high/low) | P value |
---|---|---|---|
All patients | 166 | 47.0%/15.1% | 0.006 |
Luminal (non HER2+) | 65 | 33.8%/18.5% | 0.412 |
HER2+ | 34 | 38.2%/17.6% | 0.294 |
TNBC | 67 | 64.2%/10.4% | 0.003 |
TNBC = 67 | |||
MDM2 high expression | 45 | 71.1%/11.1% | < 0.001 |
MDM2 low expression | 22 | 50.0%/9.10% | 0.005 |
Variable | pCR rate (%) | Univariate analysis P value | Multivariate analysis P value |
---|---|---|---|
FKBP12 (high/low) | 47.0/15.1 | 0.002 | 0.004 |
MDM2 (high/low) | 21.4/33.8 | 0.038 | 0.065 |
Size (pT1-2/pT3-4) | 27.2/29.4 | 0.484 | 0.507 |
Histological grade (I–II/III) | 24.6/30.8 | 0.371 | 0.438 |
LN metastasis (posi/neg) | 28.2/26.2 | 0.673 | 0.328 |
NPI (< 3.4/≥ 3.4) | 21.4/32.4 | 0.039 | 0.107 |
MIB (< 10%/≥ 10%) | 22.8/31.5 | 0.071 | 0.337 |
ERα (posi/neg) | 22.5/36.1 | 0.023 | 0.045 |
PR (posi/neg) | 19.7/35.4 | 0.028 | 0.047 |
HER2 amplification (posi/neg) | 29.7/26.3 | 0.336 | 0.503 |
Molecular subtype (TNBC/others) | 41.8/17.2 | 0.003 | 0.035 |
Variable | pCR rate (%) | Univariate analysis P value | Multivariate analysis P value |
---|---|---|---|
FKBP12 (high/low) | 64.2/10.4 | 0.003 | 0.009 |
MDM2 (high/low) | 19.4/35.8 | 0.042 | 0.105 |
Size (pT1-2/pT3-4) | 41.5/43.2 | 0.642 | 0.466 |
Histological grade (I–II/III) | 39.5/44.7 | 0.407 | 0.637 |
LN metastasis (posi/neg) | 45.3/39.1 | 0.373 | 0.578 |
NPI (< 3.4/≥ 3.4) | 38.3/46.4 | 0.175 | 0.233 |
MIB (< 10%/≥ 10%) | 36.4/52.8 | 0.043 | 0.153 |