Background
Methods
Study subjects
Treatment for TC
Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)
SNP | Primer sequence |
---|---|
rs2736100 | Forward: 5′-CCCCACAAGCTAAGCATTAT-3′ |
Reverse: 5′-GAAGAACCACGCAAAGGAC-3′ | |
rs2736098 | Forward: 5′-CCGTGGTTTCTGTGTGGTGT-3′ |
Reverse: 5′-CCCCTGAGGAGTAGAGGAAGT-3′ |
Pyrosequencing
Primer | Primer sequence |
---|---|
TERTMF | 5′-ATGATGTGGAGGTTTTGGGAATAG-3′ |
TERTMR | 5′-Biotin-CCCAACCTAAAAACAACCCTAAT-3′ |
TERTS | 5′-GGAGGTTTTGGGAATAG-3′ |
Follow-up
Statistical analysis
Results
Baseline characteristics of TC patients in the recurrence and non-recurrence groups
Characteristic | Non-recurrence group | Recurrence group |
χ
2/t
|
P
|
---|---|---|---|---|
(n = 237) | (n = 75) | |||
Gender | ||||
Male | 67 (28.3) | 24 (32.0) | 0.69 | 0.467 |
Female | 170 (71.7) | 51 (68.0) | ||
Mean age (years) | 42.58 ± 8.40 | 45.04 ± 10.53 | 1.26 | 0.211 |
Pathological type | ||||
Papillary thyroid carcinoma | 220 (92.8) | 41 (54.7) | 72.13 | <0.0001 |
Follicular thyroid carcinoma | 10 (4.2) | 7 (9.3) | ||
Medullary thyroid carcinoma | 5 (2.1) | 10 (13.3) | ||
Undifferentiated carcinoma | 2 (0.8) | 17 (22.7) | ||
Tumor stage | ||||
I | 174 (73.4) | 10 (13.3) | 98.67 | < 0.0001 |
II | 25 (10.5) | 20 (26.7) | ||
III | 35 (14.8) | 29 (38.7) | ||
IV | 3 (1.3) | 16 (21.3) |
Genotype determination of hTERT rs2736100 and rs2736098
Allele frequency and genotype distribution of hTERT rs2736100 and rs2736098 between the recurrence and non-recurrence groups
Non-recurrence group | Recurrence group |
P
| OR (95%CI) | |
---|---|---|---|---|
(n = 237) | (n = 75) | |||
rs2736100 | ||||
TT genotype | 54 (22.8) | 7 (9.3) | Ref | |
TG genotype | 133 (56.1) | 39 (52.0) | 0.059 | 0.44 (0.19–1.05) |
GG genotype | 50 (21.1) | 29 (38.7) | 0.001 | 4.47 (1.80–11.12) |
T allele | 241 (50.8) | 53 (35.3) | Ref | |
G allele | 233 (49.2) | 97 (64.7) | 0.001 | 1.89 (1.29–2.77) |
rs2736098 | ||||
GG genotype | 135 (57.0) | 46 (61.3) | Ref | |
GA genotype | 86 (36.3) | 26 (34.7) | 0.671 | 0.89 (0.51–1.54) |
AA genotype | 16 (6.8) | 3 (4.0) | 0.353 | 0.55 (0.15–1.98) |
G allele | 356 (75.1) | 118 (78.7) | Ref | |
A allele | 118 (24.9) | 32 (21.3) | 0.373 | 0.82 (0.53–1.27) |
Correlations of hTERT rs2736100 polymorphism with baseline characteristics of TC patients in the recurrence and non-recurrence groups
Characteristic | Non-recurrence group | Recurrence group | ||||
---|---|---|---|---|---|---|
TT + TG (%) | GG (%) |
P
| TT + TG (%) | GG (%) |
P
| |
187 | 50 | 46 | 29 | |||
Tumor size | ||||||
<20 mm | 127 (67.9) | 30 (60.0) | 0.293 | 31 (67.4) | 16 (55.2) | 0.287 |
≥20 mm | 60 (32.1) | 20 (40.0) | 15 (32.6) | 13 (44.8) | ||
Multicentricity | ||||||
Yes | 115 (61.5) | 28 (56.0) | 0.532 | 27 (58.7) | 9 (31.0) | 0.02 |
No | 72 (38.5) | 14 (28.0) | 19 (41.3) | 20 (69.0) | ||
Extrathyroidal invasion | ||||||
Yes | 66 (35.3) | 22 (44.0) | 0.258 | 11 (23.9) | 22 (75.9) | <0.001 |
No | 121 (64.7) | 28 (56.0) | 35 (76.1) | 7 (24.1) | ||
Lymph node metastasis | ||||||
Yes | 33 (17.6) | 11 (22.0) | 0.482 | 25 (54.3) | 23 (79.3) | 0.028 |
No | 154 (82.4) | 39 (78.0) | 21 (45.7) | 6 (20.7) | ||
Pathological type | ||||||
Papillary thyroid carcinoma | 173 (92.5) | 47 (94.0) | 0.907 | 27 (58.7) | 10 (34.5) | 0.004 |
Follicular thyroid carcinoma | 8 (4.3) | 2 (4.0) | 5 (10.9) | 2 (6.9) | ||
Medullary thyroid carcinoma | 4 (2.1) | 1 (2.0) | 8 (17.4) | 2 (6.9) | ||
Undifferentiated carcinoma | 2 (1.1) | 0 | 6 (13.0) | 15 (51.7) | ||
Tumor stage | ||||||
I + II | 156 (83.4) | 43 (86.0) | 0.659 | 21 (45.7) | 9 (31.0) | |
III + IV | 31 (16.6) | 7 (14.0) | 25 (54.3) | 20 (69.0) | 0.208 |
DNA methylation level in the promoter region of hTERT gene in the recurrence and non-recurrence groups
Group | CpG 1 | CpG 2 | CpG 3 | CpG 4 | CpG 5 | Methylation levels | Mean value |
P
|
---|---|---|---|---|---|---|---|---|
Non-recurrence group | 16% | 17% | 18% | 9% | 10% | Low | 14.0% | <0.001 |
Recurrence group | 25% | 31% | 21% | 39% | 38% | High | 30.8% |
Correlations of DNA methylation level in the promoter region of hTERT gene with baseline characteristics of TC patients in the recurrence and non-recurrence groups
Characteristic | Non-recurrence group | Recurrence group | ||||
---|---|---|---|---|---|---|
Hypomethylation (%) | Hypermethylation (%) |
P
| Hypomethylation (%) (n = 22) | Hypomethylation (%) (n = 53) |
P
| |
Tumor size | ||||||
<20 mm | 112 (70.0) | 45 (58.4) | 0.078 | 12 (54.5) | 19 (35.8) | |
≥20 mm | 48 (30.0) | 32 (41.6) | 10 (45.5) | 34 (64.2) | 0.134 | |
Multicentricity | ||||||
Yes | 102 (63.8) | 41 (53.2) | 0.122 | 9 (40.9) | 27 (50.9) | |
No | 58 (36.3) | 36 (46.8) | 13 (59.1) | 26 (49.1) | 0.428 | |
Extrathyroidal invasion | ||||||
Yes | 57 (35.6) | 31 (40.3) | 0.489 | 8 (36.4) | 25 (47.2) | 0.391 |
No | 103 (64.4) | 46 (59.7) | 14 (63.6) | 28 (52.8) | ||
Lymph node metastasis | ||||||
Yes | 29 (18.1) | 15 (19.5) | 0.802 | 7 (31.8) | 41 (77.4) | <0.001 |
No | 131 (81.9) | 62 (80.5) | 15 (68.2) | 12 (22.6) | ||
Pathological type | ||||||
Papillary thyroid carcinoma | 147 (91.9) | 73 (94.8) | 0.707 | 12 (54.5) | 25 (47.2) | |
Follicular thyroid carcinoma | 7 (4.4) | 3 (3.9) | 2 (9.1) | 5 (9.4) | ||
Medullary thyroid carcinoma | 4 (2.5) | 1 (1.3) | 2 (9.1) | 8 (15.1) | ||
Undifferentiated carcinoma | 2 (1.3) | 0 (0.0) | 6 (27.3) | 15 (28.3) | 0.897 | |
Tumor stage | ||||||
I + II | 135 (84.4) | 64 (83.1) | 0.805 | 14 (63.6) | 16 (30.2) | |
III + IV | 25 (15.6) | 13 (16.9) | 8 (36.4) | 37 (69.8) | 0.007 |