Table 1. Patient characteristics and distribution of SLC39A6 rs1050631 genotypes
Variables
|
Patients
N (%)
|
SLC39A6 rs1050631 genotype (N)
|
P*
|
Pdominant a
|
Padditive b
|
TT
|
CT
|
CC
|
Total
|
512
|
20(3.9%)
|
139(27.1%)
|
353(69.0%)
|
|
|
|
Age
|
|
|
|
|
0.26
|
0.25
|
0.14
|
<60
|
235(45.9)
|
6
|
61
|
168
|
|
|
|
≥60
|
277(54.1)
|
14
|
78
|
185
|
|
|
|
Gender
|
|
|
|
|
0.38
|
0.18
|
0.25
|
Male
|
383(74.8)
|
15
|
110
|
258
|
|
|
|
Female
|
129(25.2)
|
5
|
29
|
95
|
|
|
|
Grade of differentiation
|
|
|
|
|
0.85
|
0.93
|
0.86
|
Well differentiated
|
21(4.1)
|
1
|
6
|
14
|
|
|
|
Moderately differentiated
|
263(51.4)
|
9
|
75
|
179
|
|
|
|
Poorly differentiated
|
228(44.5)
|
10
|
58
|
160
|
|
|
|
Gross findings
|
|
|
|
|
0.79
|
0.96
|
0.81
|
Apophysis
|
35(6.8)
|
2
|
9
|
24
|
|
|
|
Invasion
|
477(93.2)
|
18
|
130
|
329
|
|
|
|
Lauren classification
|
|
|
|
|
0.65
|
0.45
|
0.22
|
Intestinal type
|
420(82.1)
|
18
|
119
|
283
|
|
|
|
Diffuse type
|
81(15.8)
|
2
|
18
|
61
|
|
|
|
Uncertain type
|
11(2.1)
|
0
|
2
|
9
|
|
|
|
Tumor location
|
|
|
|
|
0.32
|
0.38
|
0.42
|
Upper third
|
179(35.0)
|
4
|
52
|
123
|
|
|
|
Middle third
|
78(15.2)
|
4
|
16
|
58
|
|
|
|
Lower third
|
252(49.2)
|
12
|
69
|
171
|
|
|
|
Total
|
3(0.6)
|
0
|
2
|
1
|
|
|
|
Tumor size
|
|
|
|
|
0.99
|
0.88
|
0.92
|
≥5cm
|
210(41.0)
|
8
|
58
|
144
|
|
|
|
<5cm
|
302(59.0)
|
12
|
81
|
209
|
|
|
|
Survival
|
|
|
|
|
0.002
|
<0.001
|
<0.001
|
Alive
|
182(35.5)
|
4
|
35
|
143
|
|
|
|
Dead
|
330(64.5)
|
16
|
104
|
210
|
|
|
|
Recurrence
|
|
|
|
|
0.004
|
<0.001
|
<0.001
|
Yes
|
334(65.2)
|
16
|
104
|
214
|
|
|
|
No
|
178(34.8)
|
4
|
35
|
139
|
|
|
|
Distant metastasis
|
|
|
|
|
0.43
|
0.87
|
0.81
|
Yes
|
63(12.3)
|
4
|
15
|
44
|
|
|
|
No
|
449(87.7)
|
16
|
124
|
309
|
|
|
|
Lymph node metastasis
|
|
|
|
|
0.12
|
0.21
|
0.08
|
Yes
|
398(77.7)
|
19
|
110
|
269
|
|
|
|
No
|
114(22.3)
|
1
|
29
|
84
|
|
|
|
Chemotherapy
|
|
|
|
|
|
|
|
Yes
|
329(64.3)
|
13
|
93
|
223
|
0.61
|
0.78
|
0.69
|
No
|
183(35.7)
|
7
|
46
|
130
|
|
|
|
|
|
|
|
|
|
|
|
|
|
* The χ2 test was used to calculate p values. Fisher’s exact test was used in the analysis of contingency tables when sample size was smaller than 5.
a Logistic regression analysis for the dominant model (TT+CT vs CC) was used to calculate the p value.
b Logistic regression analysis for the log-additive model was used to calculate the p value by comparing the effect of each additional variant allele.
Table 2 A. Chi-square partitioning of the association between SLC39A6 rs1050631 and recurrence
SNP
|
Genotype
|
Recurrence
|
Case-totala
|
Case-caseb
|
No(n)/ Yes(n)
|
Recurrence(%)
|
p<0.00833
|
p<0.0125
|
Total(n=512)
|
|
178/334
|
65.2
|
|
|
rs1050631
|
CC
|
139/214
|
60.6
|
0.167
|
1vs2:0.003
|
|
CT
|
35/104
|
74.8
|
0.033
|
2vs3:0.784
|
|
TT
|
4/16
|
80.0
|
0.0578
|
1vs3:0.100
|
aCase–total: p value for the comparison between the group with the indicated genotype and the total group of 512 samples;
bCase–case: p value for the pairwise comparisons between groups with different genotypes.
Table 2 B. Associations between SLC39A6 rs1050631 genotypes and recurrence
SNP
|
Genotype
|
Recurrence
|
Unadjusted
|
|
|
Adjusteda
|
|
|
|
No(n)/Yes(n)
|
HR
|
95%CI
|
p
|
HR
|
95%CI
|
p
|
Total (n=512)
|
|
178/334
|
|
|
|
|
|
|
rs1050631
|
CC
|
139/214
|
1(reference)
|
1(reference)
|
|
CT
|
35/104
|
1.813
|
1.09-3.01
|
0.022
|
1.379
|
1.090-1.744
|
0.007
|
|
|
TT
|
4/16
|
1.400
|
1.11-1.77
|
0.005
|
1.441
|
0.864-2.403
|
0.161
|
|
|
CT+TT
|
39/120
|
1.440
|
1.15-1.80
|
0.001
|
1.387
|
1.108-1.735
|
0.004
|
|
|
|
|
|
|
|
|
|
|
|
|
|
The bold values indicate p value < 0.05.
aData were calculated using multivariable Cox models that adjusted for gender, age and lymph node metastasis.
Abbreviations: HR, hazard ratio; CI, Confidence Interval.
Table 3. Associations between SLC39A6 rs1050631 genotypes and recurrence, after stratification by sex, age, tumor size, differentiation grade or chemotherapy status
SNP
|
Genotype
|
Gender
|
Male
|
Female
|
Recurrence
|
Recurrence
|
No(n)/Yes(n)
|
HR
|
95%CI
|
pa
|
No(n)/Yes(n)
|
HR
|
95%CI
|
pa
|
Total
(n=512)
|
|
134/249
|
|
|
|
44/85
|
|
|
|
CC
|
105/153
|
1(reference)
|
34/61
|
1(reference)
|
CT
|
26/84
|
1.564
|
1.197-2.044
|
0.001
|
9/20
|
0.951
|
0.572-1.581
|
0.846
|
TT
|
3/12
|
1.424
|
0.788-2.573
|
0.241
|
1/4
|
1.456
|
0.524-4.048
|
0.471
|
CT+TT
|
29/96
|
1.545
|
1.196-1.996
|
0.001
|
10/24
|
1.008
|
0.626-1.624
|
0.973
|
SNP
|
Genotype
|
Age
|
<60
|
≥60
|
Recurrence
|
Recurrence
|
No(n)/Yes(n)
|
HR
|
95%CI
|
pb
|
No(n)/Yes(n)
|
HR
|
95%CI
|
pb
|
Total
(n=512)
|
|
90/145
|
|
|
|
88/189
|
|
|
|
CC
|
70/98
|
1(reference)
|
69/116
|
1(reference)
|
CT
|
18/43
|
1.242
|
0.866-1.782
|
0.238
|
17/61
|
1.512
|
1.108-2.065
|
0.009
|
TT
|
2/4
|
1.127
|
0.414-3.071
|
0.814
|
2/12
|
1.620
|
0.890-2.948
|
0.115
|
CT+TT
|
20/47
|
1.232
|
0.868-1.747
|
0.243
|
19/73
|
1.529
|
1.139-2.052
|
0.005
|
SNP
|
Genotype
|
Tumor Size
|
<5cm
|
≥5cm
|
Recurrence
|
Recurrence
|
No(n)/Yes(n)
|
HR
|
95%CI
|
pc
|
No(n)/Yes(n)
|
HR
|
95%CI
|
pc
|
Total
(n=512)
|
|
113/189
|
|
|
|
65/145
|
|
|
|
CC
|
84/125
|
1(reference)
|
55/89
|
1(reference)
|
CT
|
27/54
|
1.131
|
0.820-1.558
|
0.453
|
8/50
|
1.807
|
1.269-2.575
|
0.001
|
TT
|
2/10
|
1.333
|
0.696-2.554
|
0.386
|
2/6
|
1.657
|
0.720-3.811
|
0.235
|
CT+TT
|
29/64
|
1.158
|
0.856-1.567
|
0.341
|
10/56
|
1.789
|
1.273-2.516
|
0.001
|
SNP
|
Genotype
|
Histologic grade
|
Poorly differentiated
|
Moderately differentiated
|
Well differentiated
|
Recurrence
|
Recurrence
|
Recurrence
|
No(n)/Yes(n)
|
HR
|
95%CI
|
pc
|
No(n)/
Yes(n)
|
HR
|
95%CI
|
pc
|
No(n)/Yes(n)
|
HR
|
95%CI
|
pc
|
Total
(n=512)
|
|
70/158
|
|
103/160
|
|
5/16
|
|
CC
|
48/112
|
1(reference)
|
87/92
|
1(reference)
|
4/10
|
1(referrence)
|
CT
|
20/38
|
1.052
|
0.73-1.52
|
0.789
|
14/61
|
1.830
|
1.32-2.54
|
<0.001
|
1/5
|
1.827
|
0.54-6.22
|
0.335
|
TT
|
2/8
|
1.468
|
0.71-3.03
|
0.298
|
2/7
|
1.421
|
0.65-3.10
|
0.376
|
0/1
|
NAd
|
NAd
|
NAd
|
CT+TT
|
22/46
|
1.107
|
0.78-1.56
|
0.565
|
16/68
|
1.780
|
1.29-2.45
|
<0.001
|
1/6
|
2.131
|
0.67-6.78
|
0.200
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
SNP
|
Genotype
|
Chemotherapy
|
|
|
|
Yes
|
No
|
|
|
Recurrence
|
Recurrence
|
|
|
No(n)/Yes(n)
|
HR
|
95%CI
|
pc
|
No(n)/Yes(n)
|
HR
|
95%CI
|
pc
|
Total
|
|
128/201
|
|
|
|
50/131
|
|
|
|
(n=512)
|
CC
|
95/128
|
1(reference)
|
|
|
44/84
|
1(reference)
|
|
|
|
CT
|
29/64
|
1.27
|
0.94-1.72
|
0.981
|
6/40
|
1.79
|
1..21-2.63
|
0.003
|
|
TT
|
4/9
|
1.01
|
0.51-2.01
|
0.571
|
0/7
|
2.78
|
1.26-6.13
|
0.012
|
|
CT+TT
|
33/73
|
1.24
|
0.92-1.65
|
0.153
|
6/47
|
1.89
|
1.30-2.73
|
<0.001
|
The bold values indicate p value < 0.05.
aData were calculated using a multivariable Cox model that adjusted for age and lymph node metastasis.
bData were calculated using a multivariable Cox model that adjusted for gender and lymph node metastasis.
cData were calculated using a multivariable Cox model that adjusted for gender, age and lymph node metastasis.
dData were not available.
Abbreviations: HR, hazard ratio; CI, Confidence Interval.
Table 4 A Chi-square partitioning of the association between SLC39A6 rs1050631 and survival
SNP
|
Genotype
|
Survival
|
Case-totala
|
Case-caseb
|
Ali(n)/ Dea(n)
|
Death(%)
|
p<0.00833
|
P<0.0125
|
Total(n=512)
|
|
182/330
|
64.5
|
|
|
rs1050631
|
CC
|
143/210
|
59.5
|
0.125
|
1vs2:0.001
|
|
CT
|
35/104
|
74.8
|
0.021
|
2vs3:0.784
|
|
TT
|
4/16
|
80.0
|
0.231
|
1vs3:0.098
|
aCase–total: p value for the comparison between the group with the indicated genotype and the total group of 512 samples;
bCase–case: p value for the pairwise comparison between groups with different genotypes.
Table 4 B Associations between SLC39A6 rs1050631 genotypes and survival
SNP
|
Genotype
|
Survival
|
Unadjusted
|
|
|
Adjusteda
|
|
|
Ali(n)/Dea(n)
|
HR
|
95%CI
|
p
|
HR
|
95%CI
|
p
|
Total(n=512)
|
|
182/330
|
|
|
|
|
|
|
rs1050631
|
CC
|
143/210
|
1(reference)
|
1(reference)
|
|
CT
|
35/104
|
1.440
|
1.138-1.823
|
0.002
|
1.416
|
1.119-1.792
|
0.004
|
|
TT
|
4/16
|
1.947
|
1.170-3.239
|
0.010
|
1.523
|
0.912-2.542
|
0.107
|
|
CT+TT
|
39/120
|
1.492
|
1.192-1.868
|
<0.001
|
1.429
|
1.141-1.790
|
0.002
|
The bold values indicate p value < 0.05.
aData were calculated using a multivariable Cox model that adjusted for gender, age and lymph node metastasis.
Abbreviations: HR, hazard ratio; CI, Confidence Interval.
Table 5 Associations between SLC39A6 rs1050631 genotypes with survival, after stratification by sex, age, tumor size, differentiation grade or chemotherapy status
SNP
|
Genotype
|
Gender
|
|
|
Male
|
Female
|
|
|
Ali(n)/Dea(n)
|
HR
|
95%CI
|
pa
|
Ali(n)/Dea(n)
|
HR
|
95%CI
|
pa
|
Total
(n=512)
|
|
138/245
|
|
|
|
44/85
|
|
|
|
CC
|
109/149
|
1(reference)
|
34/61
|
1(reference)
|
CT
|
26/84
|
1.601
|
1.224-2.093
|
0.001
|
9/20
|
0.985
|
0.592-1.640
|
0.954
|
TT
|
3/12
|
1.486
|
0.822-2.688
|
0.190
|
1/4
|
1.654
|
0.595-4.600
|
0.335
|
CT+TT
|
29/96
|
1.586
|
1.226-2.051
|
<0.001
|
10/24
|
1.055
|
0.654-1.701
|
0.826
|
SNP
|
Genotype
|
Age
|
|
|
<60
|
≥60
|
|
|
Ali(n)/Dea(n)
|
HR
|
95%CI
|
pb
|
Ali(n)/Dea(n)
|
HR
|
95%CI
|
pb
|
Total
(n=512)
|
|
95/140
|
|
|
|
87/190
|
|
|
|
CC
|
74/94
|
1(reference)
|
69/116
|
1(reference)
|
CT
|
19/42
|
1.282
|
0.889-1.847
|
0.183
|
16/62
|
1.531
|
1.123-2.087
|
0.007
|
TT
|
2/4
|
1.290
|
0.473-3.517
|
0.618
|
2/12
|
1.605
|
0.880-2.926
|
0.123
|
CT+TT
|
21/46
|
1.282
|
0.900-1.827
|
0.169
|
18/74
|
1.542
|
1.150-2.068
|
0.004
|
SNP
|
Genotype
|
Tumor Size
|
|
|
<5cm
|
≥5cm
|
|
|
Ali(n)/Dea(n)
|
HR
|
95%CI
|
pc
|
Ali(n)/Dea(n)
|
HR
|
95%CI
|
pc
|
Total
(n=512)
|
|
116/186
|
|
|
|
66/144
|
|
|
|
CC
|
86/123
|
1(reference)
|
57/87
|
1(reference)
|
CT
|
28/53
|
1.141
|
0.826-1.577
|
0.423
|
7/51
|
1.928
|
1.355-2.742
|
<0.001
|
TT
|
2/10
|
1.433
|
0.747-2.749
|
0.279
|
2/6
|
1.726
|
0.750-3.974
|
0.200
|
CT+TT
|
30/63
|
1.179
|
0.869-1.600
|
0.289
|
9/57
|
1.904
|
1.355-2.675
|
<0.001
|
|
|
|
|
|
|
|
|
|
|
|
|
SNP
|
Genotype
|
Histologic grade
|
|
|
Poorly differentiated
|
Moderately differentiated
|
Well differentiated
|
|
|
Ali(n)/Dea(n)
|
HR
|
95%CI
|
pc
|
Ali(n)/Dea(n)
|
HR
|
95%CI
|
pc
|
Ali(n)/Dea(n)
|
HR
|
95%CI
|
pc
|
Total
(n=512)
|
|
75/153
|
|
|
|
102/161
|
|
|
|
5/16
|
|
|
|
|
CC
|
54/106
|
1(reference)
|
85/94
|
1(reference)
|
4/10
|
1(referrence)
|
CT
|
19/39
|
1.209
|
0.84-1.75
|
0.315
|
15/60
|
1.735
|
1.25-2.41
|
0.001
|
1/5
|
1.731
|
0.48-6.26
|
0.403
|
TT
|
2/8
|
1.479
|
0.72-3.06
|
0.291
|
2/7
|
1.482
|
0.68-3.23
|
0.322
|
0/1
|
NAd
|
NAd
|
NAd
|
CT+TT
|
21/47
|
1.248
|
0.88-1.76
|
0.209
|
17/67
|
1.706
|
1.24-2.35
|
0.001
|
1/6
|
2.192
|
0.66-7.34
|
0.203
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
SNP
|
Genotype
|
Chemotherapy
|
|
|
Yes
|
No
|
|
|
Ali(n)/Dea(n)
|
HR
|
95%CI
|
pc
|
Ali(n)/Dea(n)
|
HR
|
95%CI
|
pc
|
Total
(n=512)
|
|
135/194
|
|
|
|
47/136
|
|
|
|
CC
|
102/121
|
1(reference)
|
41/89
|
1(reference)
|
CT
|
29/64
|
1.40
|
1.03-1.89
|
0.032
|
6/40
|
1.66
|
1.13-2.43
|
0.010
|
TT
|
4/9
|
1.40
|
0.58-2.30
|
0.693
|
0/7
|
2.47
|
1.12-5.46
|
0.025
|
CT+TT
|
33/73
|
1.36
|
1.01-1.83
|
0.040
|
6/47
|
1.74
|
1.20-2.53
|
0.004
|
The bold values indicate p value < 0.05.
aData were calculated using a multivariable Cox model that adjusted for age and lymph node metastasis.
bData were calculated using a multivariable Cox model that adjusted for gender and lymph node metastasis.
cData were calculated using a multivariable Cox model that adjusted for gender, age and lymph node metastasis.
dData were not available.
Abbreviations: HR, hazard ratio; CI, Confidence Interval.
Table 6 Chi-square partitioning to test the association between SLC39A6 rs1050631 and SLC39A6 expression detected by immunohistochemistry
SNP
|
Genotype
|
expression of SLC39A6
|
Case-totala
|
Case-caseb
|
Low(n)/ High(n)
|
High expression rate (%)
|
p<0.00833
|
p<0.0125
|
Total(n=198)
|
|
112/86
|
43.4
|
|
|
rs1050631
|
CC
|
61/31
|
33.7
|
0.116
|
1vs2:0.005
|
|
CT
|
42/50
|
54.3
|
0.083
|
2vs3:0.422
|
|
TT
|
8/6
|
42.9
|
0.966
|
1vs3:0.712
|
aCase–total: p value for the comparison between the group with the indicated genotype and the total group of 198 samples;
bCase–case: p value for the pairwise comparisons between groups with different genotypes.