Introduction
Patients and methods
Patients
Methods
Results
Associations with clinicopathological factors
Associations of miR-26a, miR-101, CCNE1 and CDC2 levels with clinicopathological factors | ||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Clinicopathological factors |
N
| % | miR-26a | miR-101 |
N
| % | CCNE1 |
N
| % | CDC2 | ||||||||
Median | iqra
|
P* | Median | iqra
|
P* | Median | iqra
|
P* | Median | iqra
|
P* | |||||||
Total | 235 [65] | 100 [28] | 0.99 | 0.41 | 1.03 | 0.81 | 226 | 100 | 0.03 | 0.03 | 230 | 100 | 9.94 | 7.11 | ||||
Age in categories, year | 0.21b
| 0.09b
| 0.60b
| 0.40b
| ||||||||||||||
<40 | 12 [2] | 5 [3] | 1.04 | 0.66 | 1.19 | 0.90 | 12 | 5 | 0.03 | 0.03 | 12 | 5 | 9.41 | 0.90 | ||||
41–55 | 75 [22] | 32 [34] | 0.95 | 0.38 | 0.93 | 0.55 | 73 | 32 | 0.03 | 0.03 | 75 | 33 | 11.09 | 0.56 | ||||
56–70 | 89 [26] | 38 [40] | 1.01 | 0.38 | 1.04 | 0.87 | 84 | 37 | 0.03 | 0.03 | 89 | 39 | 9.42 | 0.87 | ||||
>70 | 59 [15] | 25 [23] | 1.05 | 0.45 | 1.25 | 0.78 | 57 | 25 | 0.03 | 0.03 | 54 | 23 | 10.05 | 0.78 | ||||
Menopausal status | 0.07c
| 0.036c
| 0.07c
| 0.14c
| ||||||||||||||
Pre menopausal | 56 [17] | 24 [26] | 0.95 | 0.40 | 0.95 | 0.66 | 55 | 24 | 0.03 | 0.03 | 56 | 24 | 11.14 | 6.31 | ||||
Postmenopausal | 179 [48] | 76 [74] | 1.02 | 0.43 | 1.07 | 0.83 | 171 | 76 | 0.03 | 0.03 | 174 | 76 | 9.85 | 7.36 | ||||
Tumor size | 0.74d
| 0.12d
| 0.45d
| 0.16d
| ||||||||||||||
pT1, <2 cm | 63 [32] | 27 [49] | 0.99 | 0.46 | 0.90 | 0.54 | 60 | 27 | 0.03 | 0.03 | 62 | 27 | 10.00 | 7.46 | ||||
pT2, >2–5 cm | 140 [30] | 60 [46] | 0.97 | 0.39 | 1.10 | 0.84 | 136 | 60 | 0.03 | 0.03 | 137 | 60 | 9.81 | 7.14 | ||||
pT3, >5 cm + pT4 | 32 [3] | 14 [5] | 1.05 | 0.49 | 1.07 | 1.04 | 30 | 13 | 0.03 | 0.03 | 31 | 13 | 11.42 | 10.11 | ||||
Lymph nodes involved | 0.79d
| 0.61d
| 0.61d
| 0.76d
| ||||||||||||||
0 | 96 [64] | 44 [98] | 0.98 | 0.37 | 0.99 | 0.57 | 92 | 41 | 0.03 | 0.03 | 96 | 42 | 9.96 | 7.62 | ||||
1–3 | 55 [1] | 25 [2] | 1.02 | 0.45 | 1.03 | 0.87 | 52 | 23 | 0.03 | 0.03 | 54 | 23 | 9.97 | 6.57 | ||||
>3 | 69 [0] | 31 [0] | 0.96 | 0.47 | 1.08 | 0.87 | 67 | 30 | 0.03 | 0.03 | 68 | 30 | 9.93 | 9.36 | ||||
Grade | 0.15d
| 0.41d
| 0.13d
| 0.41d
| ||||||||||||||
Poor | 134 [33] | 57 [51] | 0.93 | 0.44 | 1.05 | 0.80 | 126 | 56 | 0.03 | 0.03 | 131 | 57 | 10.31 | 7.01 | ||||
Unknown | 72 [24] | 31 [37] | 1.05 | 0.37 | 0.99 | 0.74 | 71 | 31 | 0.02 | 0.02 | 71 | 31 | 9.55 | 5.64 | ||||
Good/moderate | 29 [8] | 12 [12] | 0.97 | 0.44 | 0.99 | 0.83 | 29 | 13 | 0.04 | 0.04 | 28 | 12 | 9.35 | 11.54 | ||||
PgR statuse,f
| <0.001b
| <0.001b
| <0.001b
| <0.001b
| ||||||||||||||
PgR low | 44 [16] | 19 [25] | 0.87 | 0.31 | 0.76 | 0.63 | 43 | 19 | 0.03 | 0.04 | 43 | 19 | 12.84 | 9.23 | ||||
PgR high | 185 [47] | 79 [75] | 1.03 | 0.42 | 1.07 | 0.78 | 177 | 78 | 0.03 | 0.03 | 181 | 79 | 9.55 | 6.98 | ||||
HER2 statusc,e,f
| 0.026c
| 0.061c
| 0.003c
| 0.026c
| ||||||||||||||
HER2 low | 197 [51] | 84 [84] | 1.02 | 0.41 | 1.03 | 0.83 | 189 | 84 | 0.03 | 0.03 | 192 | 83 | 9.75 | 7.11 | ||||
HER2 high | 34 [10] | 14 [16] | 0.88 | 0.37 | 0.81 | 0.70 | 33 | 15 | 0.05 | 0.03 | 34 | 15 | 11.63 | 8.22 | ||||
EGFR levelse,f
| 0.044b
| 0.081b
| <0.001b
| 0.50b
| ||||||||||||||
EGFR low | 118 [27] | 50 [42] | 1.02 | 0.44 | 1.07 | 0.83 | 114 | 50 | 0.03 | 0.03 | 115 | 50 | 10.06 | 7.72 | ||||
EGFR high | 117 [38] | 50 [58] | 0.97 | 0.40 | 0.99 | 0.65 | 112 | 50 | 0.03 | 0.03 | 115 | 50 [58] | 9.91 | 7.01 |
Associations with clinical benefit and time to progression
Factor of base model |
N
| % | Univariate analysis | Multivariate analysis*
| ||||
---|---|---|---|---|---|---|---|---|
HR | 95% CI |
P
| HR | 95% CI |
P
| |||
Age (year) | ||||||||
≤55 | 87 | 37 | 1.00 | 1.00 | ||||
55–70 | 89 | 38 | 0.82 | 0.60–1.11 | 0.19 | 0.71 | 0.45–1.11 | 0.13 |
>70 | 59 | 25 | 0.66 | 0.47–0.94 | 0.02 | 0.58 | 0.36–0.94 | 0.03 |
Menopausal status | ||||||||
Premenopausal | 56 | 24 | 1.00 | |||||
Postmenopausal | 179 | 76 | 0.86 | 0.63–1.17 | 0.33 | |||
Disease-free survival | ||||||||
≤1 year | 62 | 26 | 1.00 | 1.00 | ||||
1–3 years | 109 | 46 | 0.66 | 0.48–0.91 | 0.01 | 0.63 | 0.46–0.88 | 0.006 |
>3 years | 64 | 27 | 0.51 | 0.35–0.75 | <0.001 | 0.52 | 0.36–0.77 | 0.001 |
Dominant site of relapse | ||||||||
Soft tissue | 26 | 11 | 1.00 | 1.00 | ||||
Bone | 127 | 54 | 1.29 | 0.83–2.02 | 0.26 | 1.28 | 0.79–2.07 | 0.31 |
Viscera | 82 | 35 | 1.12 | 0.70–1.79 | 0.64 | 1.29 | 0.77–2.15 | 0.33 |
ER mRNA | 235 | 100 | 0.89 | 0.83–0.94 | <0.001 | 0.90 | 0.84–0.96 | 0.002 |
PgR mRNA | 235 | 100 | 0.90 | 0.84–0.96 | 0.002 | 0.91 | 0.85–0.98 | 0.02 |
Factors analyzed | Additions to base model | |||||||
---|---|---|---|---|---|---|---|---|
mi-26a | ||||||||
Continuous variable | 235 | 100 | 0.13 | 0.06–0.28 | <0.001 | 0.18 | 0.07–0.44 | <0.001 |
Low | 79 | 34 | 1.00 | 1.00 | ||||
Intermediate | 78 | 33 | 0.93 | 0.68–1.29 | 0.68 | 1.18 | 0.83–1.66 | 0.35 |
High | 78 | 33 | 0.43 | 0.31–0.61 | <0.001 | 0.52 | 0.36–0.76 | <0.001 |
miR-101 | ||||||||
Continuous variable | 235 | 100 | 0.87 | 0.70–1.07 | 0.19 | 0.90 | 0.71–1.13 | 0.37 |
EZH2 mRNA | ||||||||
Continuous variable | 235 | 100 | 1.26 | 1.06–1.51 | 0.01 | 1.28 | 1.05–1.56 | 0.02 |
Low | 79 | 34 | 1.00 | 1.00 | ||||
Intermediate | 78 | 33 | 1.58 | 1.14–2.19 | 0.006 | 1.73 | 1.23–2.44 | 0.002 |
High | 78 | 33 | 1.91 | 1.37–2.68 | <0.001 | 1.80 | 1.26–2.55 | 0.001 |
CCNE1 mRNA | ||||||||
Continuous variable | 226 | 96 | 1.27 | 1.12–1.45 | <0.001 | 1.24 | 1.06–.144 | 0.007 |
Low | 76 | 34 | 1.00 | 1.00 | ||||
Intermediate | 75 | 33 | 1.19 | 0.85–1.66 | 0.31 | 1.24 | 0.88–1.76 | 0.22 |
High | 75 | 33 | 1.87 | 1.33–2.62 | <0.001 | 1.62 | 1.11–2.35 | 0.01 |
CDC2 mRNA | ||||||||
Continuous variable | 230 | 98 | 1.53 | 1.29–1.81 | <0.001 | 1.54 | 1.27–1.87 | <0.001 |
Low | 77 | 34 | 1.00 | 1.00 | ||||
Intermediate | 77 | 33 | 1.53 | 1.09–2.13 | 0.01 | 1.52 | 1.07–2.15 | 0.02 |
High | 76 | 33 | 2.07 | 1.47–2.90 | <0.001 | 2.05 | 1.42–2.98 | <0.001 |
miR-26a & CDC2 | ||||||||
miR-26a | 230 | 98 | 0.22 | 0.09–0.52 | <0.001 | 0.27 | 0.11–0.65 | 0.004 |
CDC2 | 230 | 98 | 1.38 | 1.15–1.65 | 0.001 | 1.47 | 1.20–1.79 | <0.001 |
Pathway analysis for miR-26a and EZH2
Global testing approach—KEGG/BioCarta pathway analysis | |||
---|---|---|---|
Genes tested |
P
| Genes significant (z-score >1.96) | |
miR-26a associated pathways
| |||
Cyclins and cell cycle regulation | 18 |
0.008
|
CCNE1,CDK7,CDKN2D,CDC2
|
TPO signaling pathway | 18 | 0.018 | HRAS,THPO,RASA1 |
EZH2 associated pathways
| |||
Cell cycle G1 S check point | 21 | 0.002 | TGFB1,E2F1,ATM,SMAD4,CDC2,CCNE1,SKP2,ATR,ABL1 |
Role of BRCA1 BRCA2 and ATR in cancer susceptibility | 20 | 0.003 | FANCG,RAD51,ATM,FANCA,CHEK1,ATR,RAD9A,NBN,FANCC,BRCA1 |
Cyclins and cell cycle regulation
| 18 |
0.005
| CCNB1,E2F1,CDC2,CCNE1,CCND2 |
ATM signaling pathway | 16 | 0.011 | RAD51,ATM,NFKB1,CHEK1,GADD45A,ABL1,NBN,BRCA1 |
Spliceosomal assembly | 15 | 0.018 | SNRPD1,SNRPG,SNRPF,U2AF1,SFRS2,U2AF2,SNRPE,SNRPA1 |
Cytokines and inflammatory response | 15 | 0.019 | TGFB1,HLA-DRA,IL15,CD4,CSF1,LTA |
Cell cycle G2 M checkpoint | 21 | 0.025 | CCNB1,ATM,CDC2,PLK1,CHEK1,ATR,WEE1,GADD45A,BRCA1 |
ADPRibosylation factor | 15 | 0.029 | KDELR1,ARFGAP1,DDEF2,PSCD4,COPA,CENTD1 |
Hypoxia and p53 in the cardiovascular system | 16 | 0.038 | ATM,FHL2,CSNK1A1,GADD45A |
p38 MAPK signaling pathway | 32 | 0.044 | TGFB1,CREB1,DAXX,CDC42,DDIT3,MAPKAPK5,HMGN1,HRAS,PLA2G4A |