Background
Materials and methods
Patients and specimens
Follow-up and postoperative treatment
Tissue microarray and immunohistochemistry
Evaluation of the immunohistochemical findings
Statistical analysis
Results
Immunohistochemical features in TMA
Correlations of 5-hmC and IDH2 expression with clinicopathologic characteristics
Clinicopathological indexes | No. of patients | No. of patients | |||||
---|---|---|---|---|---|---|---|
5-hmC Low | 5-hmC High | P† | IDH2 Low | IDH2 High | P† | ||
Sex | Female | 18 | 36 |
0.007
| 28 | 26 | 0.765 |
Male | 141 | 123 | 131 | 133 | |||
Age(year) | ≤50 | 55 | 65 | 0.247 | 60 | 60 | 1.000 |
>50 | 104 | 94 | 99 | 99 | |||
HBsAg | Negative | 30 | 26 | 0.556 | 28 | 28 | 1.000 |
Positive | 129 | 133 | 131 | 131 | |||
HCV | Negative | 158 | 158 | 1.000 | 157 | 159 | 0.156 |
Positive | 1 | 1 | 2 | 0 | |||
AFP | ≤20 | 83 | 37 |
<0.001
| 58 | 62 | 0.644 |
>20 | 76 | 122 | 101 | 97 | |||
γ-GT(U/L) | ≤54 | 87 | 81 | 0.500 | 78 | 90 | 0.178 |
>54 | 72 | 78 | 81 | 69 | |||
Liver cirrhosis | No | 32 | 26 | 0.384 | 23 | 35 | 0.081 |
Yes | 127 | 133 | 136 | 124 | |||
Tumor number | Single | 131 | 134 | 0.652 | 134 | 131 | 0.652 |
Multiple | 28 | 25 | 25 | 28 | |||
Tumor size(cm) | ≤5 | 97 | 108 | 0.197 | 99 | 106 | 0.412 |
>5 | 62 | 51 | 60 | 53 | |||
Tumor encapsulation | Complete | 94 | 88 | 0.496 | 93 | 89 | 0.650 |
None | 65 | 71 | 66 | 70 | |||
Microvascular invasion | Absent | 113 | 107 | 0.466 | 106 | 114 | 0.331 |
Present | 46 | 52 | 53 | 45 | |||
Tumor differentiation | I + II | 129 | 115 | 0.063 | 113 | 131 |
0.017
|
III + IV | 30 | 44 | 46 | 28 | |||
TNM stage | I | 98 | 93 | 0.567 | 93 | 98 | 0.567 |
II + III | 61 | 66 | 66 | 61 |
Association between combined 5-hmC and IDH2 expression and outcome in the training cohort
Factor | OS | TTR | ||||||
---|---|---|---|---|---|---|---|---|
Multivariate | Multivariate | |||||||
Univariate P | Hazard ratio | 95% CI | P† | Univariate P | Hazard ratio | 95% CI | P† | |
Sex (female vs. male) | 0.959 | NA | 0.083 | NA | ||||
Age, years (≤50 vs. >50) | 0.772 | NA | 0.597 | NA | ||||
HBsAg (negative vs. positive) | 0.983 | NA | 0.491 | NA | ||||
AFP, ng/ml (≤20 vs. >20) |
0.041
| 1.893 | 1.257–2.852 |
0.002
| 0.230 | NA | ||
γ-GT, U/L (≤54 vs. >54) |
0.006
| 1.619 | 1.118–2.343 |
0.011
|
0.003
| 1.547 | 1.138–2.102 |
0.005
|
Liver cirrhosis (no vs. yes) | 0.077 | NA |
0.009
| 1.824 | 1.135–2.930 |
0.013
| ||
Tumor number (single vs. multiple) |
0.003
| NS |
0.002
| 1.651 | 1.135–2.402 |
0.009
| ||
Tumor size, cm (≤5 vs. >5) |
0.009
| NS |
0.041
| NS | ||||
Tumor encapsulation (complete vs. none) | 0.261 | NA | 0.166 | NA | ||||
Microvascular invasion (no vs. yes) |
0.003
| NS |
0.001
| 1.775 | 1.287–2.448 |
<0.001
| ||
Tumor differentiation (I-II vs. III-IV) | 0.138 | NA | 0.053 | NA | ||||
TNM stage (I vs. II III) |
<0.001
| 2.048 | 1.412–2.971 |
<0.001
|
<0.001
| 1.649 | 1.134–2.397 |
0.009
|
5-hmC (low vs. high) |
<0.001
| 0.316 | 0.211–0.472 |
<0.001
|
0.001
| 0.462 | 0.335–0.636 |
<0.001
|
IDH2 (low vs. high) |
<0.001
| 0.405 | 0.275–0.594 |
<0.001
|
0.006
| 0.591 | 0.432–0.810 |
0.001
|
Combination of 5-hmC and IDH2 |
<0.001
|
<0.001
|
<0.001
|
<0.001
| ||||
I versus II |
0.002
| 3.987 | 1.890–8.413 |
<0.001
|
0.001
| 2.651 | 1.576–4.461 |
<0.001
|
I versus III |
0.002
| 3.359 | 1.607–7.025 |
0.001
|
0.003
| 2.098 | 1.247–3.530 |
0.005
|
I versus IV |
<0.001
| 8.908 | 4.215–18.825 |
<0.001
|
<0.001
| 3.891 | 2.270–6.671 |
<0.001
|