Table 1
Clinical and pathohistological characteristics of patients included in study
| Clinical and histopathological characteristics | CRC patients (N = 163) |
|---|---|
| Sex | |
| Male/female, N(%) | 92/71 (56.4/43.6) |
| Age at diagnosis | |
| Mean +/- SD | 67.28 +/- 11.44 |
| Stage of disease, N (%) | |
| I | 29 (17.8) |
| II | 81 (49.7) |
| III | 53 (32.5) |
| TNM staging | |
| Tumour, N (%) | |
| T1 | 8 (4.9) |
| T2 | 24 (14.7) |
| T3 | 120 (73.6) |
| T4 | 11 (6.7) |
| Lymph nodes, N (%) | |
| N0 | 110 (67.5) |
| N1 | 38 (23.3) |
| N2 | 15 (9.2) |
| Vital status (5.10.2015), N (%) | |
| dead | 65 (39.9) (42 due to CRC |
| progression, 23 other cause) | |
| alive | 98 (60.1) |
| Disease progression, N (%) | |
| yes | 46 (28.2) (42 dead, 4 alive) |
| no | 117 (71.8) |
| Disease progression according to stage, N (%) | |
| stage I | 5 (17.2) |
| stage II | 18 (22.2) |
| stage III | 23 (43.4) |
| Clinical characteristics, N (%) | |
| more than 12 lymph nodes resected | 70 (42.9) |
| adjuvant therapy | 41 (25.2) |
| Differentiation grade, N (%) | |
| I | 66 (40.5) |
| II | 74 (45.4) |
| III | 21 (12.9) |
| no data | 2 (1.2) |
| Perineural invasion, N (%) | |
| yes | 13 (8.0) |
| no | 150 (92.0) |
| Lymphovascular invasion, N (%) | |
| yes | 27 (16.6) |
| no | 136 (83.4) |
N = number of patients
Table 2
Primer sequences used in PCR reaction, expected sizes of products, annealing temperatures, primer concentrations and genotyping method of selected SNPs; restriction enzymes and sizes of fragments after restriction for genotyping of SNPs rs1801157 (CXCL12) and rs1990172 (MACC1)
| Gene | SNP ID | Forward and reverse primer | Product size [bp] | Annealing temperature [°C] | Primer concentration [nM] | Genotyping method |
|---|---|---|---|---|---|---|
| CXCL12 | rs1801157 A/G | GTGGGATGGGATGGTGGAG CCTCAGCTCAGGGTAGCC | 109 | 60 | 650 | RFLP |
| MACC1 | rs1990172 G/T | CAGGGAAAGAAATGGTTATTGCA GGAAAAGGAGGGAAGCATGTG | 115 | 63 | 300 | RFLP |
| MMP7 | rs11568818 A/G | TGGAGTCAATTTATGCAGCAG CGAGGAAGTATTACATCGTTATTGG | 93 | 57 | 250 | HRM |
| RAD18 | rs373572 A/G | TGTGATTAACCTAGTGGTTATTTTCTT GCATCCTAGTCTTCTCTATATTTTCG | 85 | 60 | 300 | HRM |
| Gene | SNP ID | Restriction enzyme | Size of fragments after restriction [bp] | |||
| CXCL12 | rs1801157 | MspI | AA: 109, AG: 109+62+47, GG: 62+47 | |||
| MACC1 | rs1990172 | BseGI | GG: 80+35, GT: 105+80+35, TT: 105 | |||
Table 3
Associations between selected SNPs and clinico-histopathological characteristics of patients
| TNM staging | Grade of differentiation | Perineural invasion | Lymphovascular invasion | |||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Gene/ SNP ID | Frequency | T1+T2 | T3+T4 | N0 | N1+N2 | 1+2 | 3 | No | Yes | No | Yes | |||
| CXCL12 rs1801157 | AA | 4.76 | 0.0 | 5.9 | 7.3 | 0.0 | 4.8 | 5.3 | 5.2 | 0.0 | 5.8 | 0.0 | ||
| AG | 29.25 | 31.0 | 28.8 | 26.0 | 35.3 | 27.8 | 42.1 | 29.9 | 23.1 | 27.3 | 38.5 | |||
| GG | 65.99 | 69.0 | 65.3 | 66.7 | 64.7 | 67.5 | 52.6 | 64.9 | 76.9 | 66.9 | 61.5 | |||
| Statistical analysis | 0.346 | 0.096 | 1.000 | 1.000 | 0.354 | p-value | AA vs AG+GG | |||||||
| 1.261 | 1.573 | 0.900 | 1.102 | 1.228 | OR | |||||||||
| 1.159–1.373 | 1.388–1.783 | 0.102–7.916 | 1.045–1.162 | 1.135–1.329 | 95% CI | |||||||||
| 0.828 | 1.000 | 0.303 | 0.543 | 0.656 | p-value | AA+AG vs GG | ||||||||
| 0.814 | 0.960 | 0.556 | 1.860 | 0.820 | OR | |||||||||
| 0.340–1.948 | 0.471–1.958 | 0.210–1.471 | 0.488–7.089 | 0.342–1.967 | 95% CI | |||||||||
| MACC1 rs1990172 | GG | 5.84 | 12.0 | 4.5 | 7.8 | 2.1 | 5.2 | 5.3 | 5.6 | 7.7 | 7.1 | 0.0 | ||
| GT | 36.50 | 52.0 | 33.0 | 37.8 | 34.0 | 37.9 | 26.3 | 38.7 | 15.4 | 35.4 | 41.7 | |||
| TT | 57.66 | 36.0 | 62.5 | 54.4 | 63.8 | 56.9 | 68.4 | 55.6 | 76.9 | 57.5 | 58.3 | |||
| Statistical analysis | 0.024 | 0.363 | 1.000 | 0.560 | 0.350 | p-value | GG vs GT+TT | |||||||
| 2.963 | 1.477 | 0.982 | 0.718 | 1.229 | OR | |||||||||
| 1.202–7.301 | 0.715–3.050 | 0.112–8.641 | 0.081–6.338 | 1.131–1.334 | 95% CI | |||||||||
| 0.263 | 0.160 | 0.453 | 0.237 | 1.000 | p-value | GG+GT vs TT | ||||||||
| 3.880 | 2.918 | 1.641 | 2.657 | 1.034 | OR | |||||||||
| 0.463–32.516 | 0.649–13.119 | 0.583–4.620 | 0.697–10.127 | 0.523–2.525 | 95% CI | |||||||||
| MMP7 rs11568818 | AA | 27.27 | 28.6 | 27.0 | 28.1 | 25.5 | 27.9 | 26.3 | 26.7 | 33.3 | 27.7 | 25.0 | ||
| AG | 47.55 | 64.3 | 43.5 | 52.1 | 38.3 | 47.5 | 52.6 | 48.9 | 33.3 | 51.3 | 29.2 | |||
| GG | 25.17 | 7.1 | 29.6 | 19.8 | 36.2 | 24.6 | 21.1 | 24.4 | 33.3 | 21.0 | 45.8 | |||
| Statistical analysis | 1.000 | 0.843 | 1.000 | 0.736 | 1.000 | p-value | AA vs AG+GG | |||||||
| 1.084 | 1.141 | 1.082 | 0.729 | 1.151 | OR | |||||||||
| 0.433–2.713 | 0.517–2.521 | 0.362–3.234 | 0.207–2.573 | 0.420–3.152 | 95% CI | |||||||||
| 0.014 | 0.041 | 1.000 | 0.497 | 0.018 | p-value | AA+AG vs GG | ||||||||
| 5.457 | 2.296 | 0.818 | 1.547 | 3.182 | OR | |||||||||
| 1.226–24.284 | 1.054–5.002 | 0.252–2.654 | 0.437–5.479 | 1.273–7.952 | 95% CI | |||||||||
| RAD18 rs373572 | AA | 51.59 | 46.7 | 52.8 | 48.6 | 58.0 | 50.4 | 55.0 | 50.7 | 61.5 | 51.9 | 50.0 | ||
| AG | 38.22 | 33.3 | 39.4 | 41.1 | 32.0 | 39.3 | 35.0 | 39.6 | 23.1 | 38.2 | 38.5 | |||
| GG | 10.19 | 20.0 | 7.9 | 10.3 | 10.0 | 10.4 | 10.0 | 9.7 | 15.4 | 9.9 | 11.5 | |||
| Statistical analysis | 0.685 | 0.306 | 0.812 | 0.567 | 1.000 | p-value | AA vs AG+GG | |||||||
| 0.784 | 0.685 | 0.830 | 0.643 | 1.079 | OR | |||||||||
| 0.353–1.739 | 0.348–1.348 | 0.323–2.133 | 0.201–2.058 | 0.465–2.504 | 95% CI | |||||||||
| 0.085 | 1.000 | 1.000 | 0.624 | 0.731 | p-value | AA+AG vs GG | ||||||||
| 0.342 | 0.970 | 0.960 | 1.688 | 1.184 | OR | |||||||||
| 0.113–1.030 | 0.318–2.957 | 0.201–4.580 | 0.339–8.399 | 0.312–4.488 | 95% CI | |||||||||
Survival analysis curves for different genotype groups of CRC patients according to SNP rs1990172 in gene MACC1.
Survival analysis curves for different genotype groups of CRC patients according to SNP rs373572 in gene RAD18.